LOCUS       TLCV2.       16733 bp DNA     circular SYN 13-MAY-2021
DEFINITION  LentiCRISPR v2 was modified into an all-in-one dox inducible system.
            The addition of doxycycline induces Cas9-2A-eGFP. The U6 promoter 
            drives constitutive sgRNA expression..
ACCESSION   .
VERSION     .
KEYWORDS    TLCV2.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16733)
  AUTHORS   Barger CJ, Branick C, Chee L, Karpf AR
  TITLE     Pan-Cancer Analyses Reveal Genomic Features of FOXM1 Overexpression 
            in Cancer.
  JOURNAL   Cancers (Basel). 2019 Feb 21;11(2). pii: cancers11020251. doi: 
            10.3390/cancers11020251.
  PUBMED    30795624
REFERENCE   2  (bases 1 to 16733)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 16733)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cancers
            (Basel)."; date: "2019-02-21"; pages: " 10.3390/cancers11020251"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..16733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    14..32
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    49..67
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    85..103
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    121..139
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    156..174
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    192..210
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        222..260
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     257..281
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             316..4419
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     CDS             4420..4467
                     /codon_start=1
                     /product="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4468..4491
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             4501..4554
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4555..5268
                     /label=EGFP
                     /note="enhanced GFP"
     promoter        5282..5493
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             5515..6111
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     CDS             6121..6177
                     /label=P2A
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     CDS             6178..6921
                     /label=rtTA-Advanced
                     /note="improved tetracycline-controlled transactivator"
     misc_feature    6945..7533
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             7605..7838
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7870..8094
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8140..8568
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        8582..8911
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        8959..9006
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             9025..9396
                     /label=BleoR
                     /note="antibiotic-binding protein"
     polyA_signal    9529..9662
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9699..9715)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9699..9715)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9712..9734)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9723..9739
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(9747..9777)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(9792..9813)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(9930..9947)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10101..10689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10863..11720)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(11721..11825)
                     /label=AmpR promoter
     primer_bind     complement(11900..11919)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     enhancer        12091..12470
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        12471..12673
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             12688..12868
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    12915..13040
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    13533..13766
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             13951..13995
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
     CDS             14144..14185
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    14293..14410
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        14461..14701
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        16591..16666
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    16711..16729
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 atgtcgagtt tactccctat cagtgataga gaacgatgtc gagtttactc cctatcagtg
       61 atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac
      121 tccctatcag tgatagagaa cgtatgtcga gtttatccct atcagtgata gagaacgtat
      181 gtcgagttta ctccctatca gtgatagaga acgtatgtcg aggtaggcgt gtacggtggg
      241 aggcctatat aagcagagct cgtttagtga accgtcagat cgcctggaga ataccggttc
      301 tagagcgctg ccaccatgga caagaagtac agcatcggcc tggacatcgg caccaactct
      361 gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg
      421 ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc
      481 ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg
      541 aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac
      601 agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg
      661 caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc
      721 taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat
      781 ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac
      841 cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg
      901 ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga
      961 ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat
     1021 ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac
     1081 ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg
     1141 gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac
     1201 ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc
     1261 cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg
     1321 aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc
     1381 aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc
     1441 atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga
     1501 gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac
     1561 ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac
     1621 aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg
     1681 gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc
     1741 tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg
     1801 accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac
     1861 gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga
     1921 aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc
     1981 aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc
     2041 gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac
     2101 gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt
     2161 ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg
     2221 ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga
     2281 tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc
     2341 ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag
     2401 ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc
     2461 cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag
     2521 ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag
     2581 cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag
     2641 aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc
     2701 ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac
     2761 ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac
     2821 tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag
     2881 gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc
     2941 gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga
     3001 aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc
     3061 ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg
     3121 gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg
     3181 atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg
     3241 cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc
     3301 gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg
     3361 tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag
     3421 tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc
     3481 gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat
     3541 aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg
     3601 aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac
     3661 agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac
     3721 agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag
     3781 aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag
     3841 aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc
     3901 atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc
     3961 tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc
     4021 ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa
     4081 cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag
     4141 ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac
     4201 aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg
     4261 accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg
     4321 tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg
     4381 tacgagacac ggatcgacct gtctcagctg ggaggcgaca agcgacctgc cgccacaaag
     4441 aaggctggac aggctaagaa gaagaaagat tacaaagacg atgacgataa gggatccgga
     4501 gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg cccagtgagc
     4561 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     4621 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     4681 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     4741 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     4801 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     4861 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     4921 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     4981 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     5041 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     5101 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     5161 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     5221 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agctagcatc
     5281 cgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     5341 tccggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     5401 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     5461 ctttttcgca acgggtttgc cgccagaaca cagactgcga tcgcaatgta cagtatgacc
     5521 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     5581 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     5641 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     5701 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     5761 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     5821 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     5881 tggttcctgg ccaccgtcgg agtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     5941 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     6001 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     6061 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc cggttccggc
     6121 gcaacaaact tctctctgct gaaacaagcc ggagatgtcg aagagaatcc tggaccgatg
     6181 tctagactgg acaagagcaa agtcataaac ggagctctgg aattactcaa tggtgtcggt
     6241 atcgaaggcc tgacgacaag gaaactcgct caaaagctgg gagttgagca gcctaccctg
     6301 tactggcacg tgaagaacaa gcgggccctg ctcgatgccc tgccaatcga gatgctggac
     6361 aggcatcata cccacttctg ccccctggaa ggcgagtcat ggcaagactt tctgcggaac
     6421 aacgccaagt cataccgctg tgctctcctc tcacatcgcg acggggctaa agtgcatctc
     6481 ggcacccgcc caacagagaa acagtacgaa accctggaaa atcagctcgc gttcctgtgt
     6541 cagcaaggct tctccctgga gaacgcactg tacgctctgt ccgccgtggg ccactttaca
     6601 ctgggctgcg tattggagga acaggagcat caagtagcaa aagaggaaag agagacacct
     6661 accaccgatt ctatgccccc acttctgaga caagcaattg agctgttcga ccggcaggga
     6721 gccgaacctg ccttcctttt cggcctggaa ctaatcatat gtggcctgga gaaacagcta
     6781 aagtgcgaaa gcggcgggcc gaccgacgcc cttgacgatt ttgacttaga catgctccca
     6841 gccgatgccc ttgacgactt tgaccttgat atgctgcctg ctgacgctct tgacgatttt
     6901 gaccttgaca tgctccccgg gtaactcgag cgcgttaagt cgacaatcaa cctctggatt
     6961 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     7021 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     7081 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     7141 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     7201 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     7261 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     7321 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     7381 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     7441 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     7501 cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa
     7561 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     7621 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     7681 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     7741 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     7801 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat
     7861 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     7921 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     7981 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     8041 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     8101 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     8161 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     8221 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     8281 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     8341 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     8401 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     8461 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     8521 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     8581 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     8641 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     8701 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     8761 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     8821 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     8881 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     8941 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     9001 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
     9061 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     9121 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     9181 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     9241 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     9301 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     9361 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     9421 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     9481 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     9541 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     9601 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     9661 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     9721 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     9781 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     9841 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     9901 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     9961 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
    10021 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
    10081 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
    10141 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
    10201 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
    10261 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
    10321 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
    10381 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
    10441 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
    10501 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
    10561 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
    10621 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
    10681 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
    10741 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
    10801 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
    10861 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
    10921 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
    10981 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
    11041 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
    11101 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
    11161 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
    11221 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
    11281 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
    11341 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
    11401 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
    11461 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
    11521 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
    11581 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
    11641 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
    11701 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
    11761 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
    11821 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
    11881 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
    11941 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
    12001 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
    12061 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
    12121 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
    12181 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
    12241 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
    12301 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
    12361 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
    12421 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
    12481 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
    12541 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
    12601 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
    12661 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
    12721 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
    12781 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
    12841 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
    12901 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
    12961 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
    13021 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
    13081 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
    13141 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
    13201 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
    13261 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
    13321 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
    13381 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
    13441 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
    13501 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
    13561 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
    13621 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
    13681 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
    13741 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
    13801 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
    13861 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
    13921 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
    13981 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
    14041 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
    14101 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
    14161 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
    14221 tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac aaatggcagt
    14281 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
    14341 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
    14401 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa ttaaggtacc
    14461 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
    14521 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
    14581 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
    14641 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
    14701 cgaaacaccg gagacggttg taaatgagca cacaaaatac acatgctaaa atattatatt
    14761 ctatgacctt tataaaatca accaaaatct tctttttaat aactttagta tcaataatta
    14821 gaatttttat gttccttttt gcaaactttt aataaaaatg agcaaaataa aaaaacgcta
    14881 gttttagtaa ctcgcgttgt tttcttcacc tttaataata gctactccac cacttgttcc
    14941 taagcggtca gctcctgctt caatcatttt ttgagcatct tcaaatgttc taactccacc
    15001 agctgcttta actaaagcat tgtctttaac aactgacttc attagtttaa catcttcaaa
    15061 tgttgcacct gattttgaaa atcctgttga tgttttaaca aattctaatc cagcttcaac
    15121 agctatttca caagctttca tgatttcttc ttttgttaat aaacaatttt ccataataca
    15181 tttaacaaca tgtgatccag ctgctttttt tacagctttc atgtcttcta aaactaattc
    15241 ataatttttg tcttttaatg caccaatatt taataccata tcaatttctg ttgcaccatc
    15301 tttaattgct tcagaaactt cgaatgcttt tgtagctgtt gtgcatgcac ctagaggaaa
    15361 acctacaaca tttgttattc ctacatttgt gccttttaat aattctttac aatagcttgt
    15421 tcaatatgaa ttaacacaaa ctgttgcaaa atcaaattca attgcttcat cacataattg
    15481 tttaatttca gctttcgtag catcttgttt taataatgtg tgatctatat atttgtttag
    15541 tttcattttt tctcctatat attcattttt aattttaatt ctttaataat ttcgtctact
    15601 ttaactttag cgttttgaac agattcacca acacctataa aataaatttt tagtttaggt
    15661 tcagttccac ttgggcgaac agcaaatcat gacttatctt ctaaataaaa ttttagtaag
    15721 tcttgtcctg gcatattata cattccatcg atgtagtctt caacattaac aactttaagt
    15781 ccagcaattt gagttaaggg tgttgctctc aatgatttca ttaatggttc aatttttaat
    15841 ttcttttctt ctggtttaaa attcaagttt aaagtgaaag tgtaatatgc acccatttct
    15901 ttaaataaat cttctaaata gtctactaat gttttatttt gttttttata aaatcaagca
    15961 gcctctgcta ttaatataga agcttgtatt ccatctttat ctctagctga gtcatcaatt
    16021 acatatccat aactttcttc ataagcaaaa acaaaattta atccgttatc ttcttcttta
    16081 gcaatttctc tacccattca tttaaatcca gttaaagttt ttacaatatt aactccatat
    16141 ttttcatgag cgattctatc acccaaatca cttgttacaa aacttgaata tagagccgga
    16201 ttttttggaa tgctatttaa gcgttttaga tttgataatt ttcaatcaat taaaattggt
    16261 cctgtttgat ttccatctaa tcttacaaaa tgaccatcat gttttattgc cattccaaat
    16321 ctgtcagcat ctgggtcatt cataataata atatctgcat catgtttaat accatattca
    16381 agcggtattt ttcatgcagg atcaaattct ggatttggat ttacaacatt tttaaatgtt
    16441 tcatcttcaa atgcatgctc ttcaacctca ataacgttat atcctgattc acgtaatatt
    16501 tttggggtaa atttagttcc tgttccatta actgcgctaa aaataatttt taaatctttt
    16561 ttagcttctt gctctttttt gtacgtctct gttttagagc tagaaatagc aagttaaaat
    16621 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgaattcac
    16681 tttggccgcg aatcgatatg tcgagtttac tccctatcag tgatagagaa cgt
//
