LOCUS       V001427                14982 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001427
VERSION     V001427
KEYWORDS    pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 14982)
  AUTHORS   Thakore PI, D'Ippolito AM, Song L, Safi A, Shivakumar NK, Kabadi AM,
            Reddy TE, Crawford GE, Gersbach CA
  TITLE     Highly specific epigenome editing by CRISPR-Cas9 repressors for
            silencing of distal regulatory elements.
  JOURNAL   Nat Methods. 2015 Dec;12(12):1143-9. doi: 10.1038/nmeth.3630. Epub
            2015 Oct 26.
   PUBMED   26501517
REFERENCE   2  (bases 1 to 14982)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 14982)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi:
            "10.1038/nmeth.3630"; journalName: "Nat Methods"; date: "2015-12";
            volume: "12"; issue: "12"; pages: "1143-9"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..14982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..583
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             598..778
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1443..1676
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1861..1905
                     /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             2054..2095
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2203..2320
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     protein_bind    complement(2364..2397)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_RNA        complement(2408..2483)
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     promoter        complement(2508..2748)
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     promoter        2781..3992
                     /label="UbC promoter"
                     /note="human ubiquitin C promoter"
     regulatory      4021..4030
                     /label="Kozak sequence"
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4072..4095
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             4102..4122
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             4132..8235
                     /label="dCas9"
                     /note="catalytically dead mutant of the Cas9 endonuclease
                     from the Streptococcus pyogenes Type II CRISPR/Cas system"
     CDS             8248..8268
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label="SV40 NLS"
                     /translation="PKKKRKV"
     CDS             8302..8496
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional
                     repression domain from the human zinc finger protein ZNF10
                     (Margolin et al., 1994)"
                     /label="KRAB"
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQILYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             8563..8583
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             8590..8643
                     /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             8650..9363
                     /label="EGFP"
                     /note="enhanced GFP"
     protein_bind    complement(9379..9412)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(9490..9510)
                     /label="WPRE-R"
                     /note="WPRE, reverse primer"
     CDS             complement(9908..9919)
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     primer_bind     complement(10028..10044)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     complement(10029..10045)
                     /label="pBluescriptKS"
                     /note="For pBluescript vector"
     LTR             10550..10730
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    10762..10986
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11032..11460
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        11474..11803
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        11851..11898
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             11917..12288
                     /label="BleoR"
                     /note="antibiotic-binding protein"
     polyA_signal    12421..12554
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12591..12607)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     complement(12591..12607)
                     /label="M13 Reverse"
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12604..12626)
                     /label="M13/pUC Reverse"
                     /note="In lacZ gene"
     protein_bind    12615..12631
                     /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(12639..12669)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(12684..12705)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(12822..12839)
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12993..13581)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(13755..14612)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(14613..14717)
                     /label="AmpR promoter"
     primer_bind     complement(14792..14811)
                     /label="pRS-marker"
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaataactt cgtatagcat acattatacg aagttattaa
     2401 aaaaaaagca ccgactcggt gccacttttt caagttgata acggactagc cttattttaa
     2461 cttgctattt ctagctctaa aactgagacg tccgtctccg gtgtttcgtc ctttccacaa
     2521 gatatataaa gccaagaaat cgaaatactt tcaagttacg gtaagcatat gatagtccat
     2581 tttaaaacat aattttaaaa ctgcaaacta cccaagaaat tattactttc tacgtcacgt
     2641 attttgtact aatatctttg tgtttacagt caaattaatt ccaattatct ctctaacagc
     2701 cttgtatcgt atatgcaaat atgaaggaat catgggaaat aggccctctt aattaattaa
     2761 cccgtgtcgg ctccagatct ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc
     2821 cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc
     2881 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     2941 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     3001 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     3061 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     3121 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     3181 gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg
     3241 gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga
     3301 actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac
     3361 gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga
     3421 acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
     3481 gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg
     3541 tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga
     3601 gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg
     3661 gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc
     3721 tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga
     3781 ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt
     3841 tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg
     3901 aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gactagtaaa ttgtccgcta
     3961 aattctggcc gtttttggct tttttgttag acgaagcttg ggctgcaggt cgactctaga
     4021 gccaccatgg actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag
     4081 gatgacgatg acaagatggc ccccaagaag aagaggaagg tgggccgcgg aatggacaag
     4141 aagtactcca ttgggctcgc catcggcaca aacagcgtcg gctgggccgt cattacggac
     4201 gagtacaagg tgccgagcaa aaaattcaaa gttctgggca ataccgatcg ccacagcata
     4261 aagaagaacc tcattggcgc cctcctgttc gactccgggg aaaccgccga agccacgcgg
     4321 ctcaaaagaa cagcacggcg cagatatacc cgcagaaaga atcggatctg ctacctgcag
     4381 gagatcttta gtaatgagat ggctaaggtg gatgactctt tcttccatag gctggaggag
     4441 tcctttttgg tggaggagga taaaaagcac gagcgccacc caatctttgg caatatcgtg
     4501 gacgaggtgg cgtaccatga aaagtaccca accatatatc atctgaggaa gaagcttgta
     4561 gacagtactg ataaggctga cttgcggttg atctatctcg cgctggcgca tatgatcaaa
     4621 tttcggggac acttcctcat cgagggggac ctgaacccag acaacagcga tgtcgacaaa
     4681 ctctttatcc aactggttca gacttacaat cagcttttcg aagagaaccc gatcaacgca
     4741 tccggagttg acgccaaagc aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa
     4801 aacctcatcg cacagctccc tggggagaag aagaacggcc tgtttggtaa tcttatcgcc
     4861 ctgtcactcg ggctgacccc caactttaaa tctaacttcg acctggccga agatgccaag
     4921 cttcaactga gcaaagacac ctacgatgat gatctcgaca atctgctggc ccagatcggc
     4981 gaccagtacg cagacctttt tttggcggca aagaacctgt cagacgccat tctgctgagt
     5041 gatattctgc gagtgaacac ggagatcacc aaagctccgc tgagcgctag tatgatcaag
     5101 cgctatgatg agcaccacca agacttgact ttgctgaagg cccttgtcag acagcaactg
     5161 cctgagaagt acaaggaaat tttcttcgat cagtctaaaa atggctacgc cggatacatt
     5221 gacggcggag caagccagga ggaattttac aaatttatta agcccatctt ggaaaaaatg
     5281 gacggcaccg aggagctgct ggtaaagctt aacagagaag atctgttgcg caaacagcgc
     5341 actttcgaca atggaagcat cccccaccag attcacctgg gcgaactgca cgctatcctc
     5401 aggcggcaag aggatttcta cccctttttg aaagataaca gggaaaagat tgagaaaatc
     5461 ctcacatttc ggatacccta ctatgtaggc cccctcgccc ggggaaattc cagattcgcg
     5521 tggatgactc gcaaatcaga agagaccatc actccctgga acttcgagga agtcgtggat
     5581 aagggggcct ctgcccagtc cttcatcgaa aggatgacta actttgataa aaatctgcct
     5641 aacgaaaagg tgcttcctaa acactctctg ctgtacgagt acttcacagt ttataacgag
     5701 ctcaccaagg tcaaatacgt cacagaaggg atgagaaagc cagcattcct gtctggagag
     5761 cagaagaaag ctatcgtgga cctcctcttc aagacgaacc ggaaagttac cgtgaaacag
     5821 ctcaaagaag actatttcaa aaagattgaa tgtttcgact ctgttgaaat cagcggagtg
     5881 gaggatcgct tcaacgcatc cctgggaacg tatcacgatc tcctgaaaat cattaaagac
     5941 aaggacttcc tggacaatga ggagaacgag gacattcttg aggacattgt cctcaccctt
     6001 acgttgtttg aagataggga gatgattgaa gaacgcttga aaacttacgc tcatctcttc
     6061 gacgacaaag tcatgaaaca gctcaagagg cgccgatata caggatgggg gcggctgtca
     6121 agaaaactga tcaatgggat ccgagacaag cagagtggaa agacaatcct ggattttctt
     6181 aagtccgatg gatttgccaa ccggaacttc atgcagttga tccatgatga ctctctcacc
     6241 tttaaggagg acatccagaa agcacaagtt tctggccagg gggacagtct tcacgagcac
     6301 atcgctaatc ttgcaggtag cccagctatc aaaaagggaa tactgcagac cgttaaggtc
     6361 gtggatgaac tcgtcaaagt aatgggaagg cataagcccg agaatatcgt tatcgagatg
     6421 gcccgagaga accaaactac ccagaaggga cagaagaaca gtagggaaag gatgaagagg
     6481 attgaagagg gtataaaaga actggggtcc caaatcctta aggaacaccc agttgaaaac
     6541 acccagcttc agaatgagaa gctctacctg tactacctgc agaacggcag ggacatgtac
     6601 gtggatcagg aactggacat caatcggctc tccgactacg acgtggatgc catcgtgccc
     6661 cagtcttttc tcaaagatga ttctattgat aataaagtgt tgacaagatc cgataaaaat
     6721 agagggaaga gtgataacgt cccctcagaa gaagttgtca agaaaatgaa aaattattgg
     6781 cggcagctgc tgaacgccaa actgatcaca caacggaagt tcgataatct gactaaggct
     6841 gaacgaggtg gcctgtctga gttggataaa gccggcttca tcaaaaggca gcttgttgag
     6901 acacgccaga tcaccaagca cgtggcccaa attctcgatt cacgcatgaa caccaagtac
     6961 gatgaaaatg acaaactgat tcgagaggtg aaagttatta ctctgaagtc taagctggtc
     7021 tcagatttca gaaaggactt tcagttttat aaggtgagag agatcaacaa ttaccaccat
     7081 gcgcatgatg cctacctgaa tgcagtggta ggcactgcac ttatcaaaaa atatcccaag
     7141 cttgaatctg aatttgttta cggagactat aaagtgtacg atgttaggaa aatgatcgca
     7201 aagtctgagc aggaaatagg caaggccacc gctaagtact tcttttacag caatattatg
     7261 aattttttca agaccgagat tacactggcc aatggagaga ttcggaagcg accacttatc
     7321 gaaacaaacg gagaaacagg agaaatcgtg tgggacaagg gtagggattt cgcgacagtc
     7381 cggaaggtcc tgtccatgcc gcaggtgaac atcgttaaaa agaccgaagt acagaccgga
     7441 ggcttctcca aggaaagtat cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa
     7501 aaagattggg accccaagaa atacggcgga ttcgattctc ctacagtcgc ttacagtgta
     7561 ctggttgtgg ccaaagtgga gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg
     7621 ctgggcatca caatcatgga gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag
     7681 gcgaaaggat ataaagaggt caaaaaagac ctcatcatta agcttcccaa gtactctctc
     7741 tttgagcttg aaaacggccg gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt
     7801 aacgagctgg cactgccctc taaatacgtt aatttcttgt atctggccag ccactatgaa
     7861 aagctcaaag ggtctcccga agataatgag cagaagcagc tgttcgtgga acaacacaaa
     7921 cactaccttg atgagatcat cgagcaaata agcgaattct ccaaaagagt gatcctcgcc
     7981 gacgctaacc tcgataaggt gctttctgct tacaataagc acagggataa gcccatcagg
     8041 gagcaggcag aaaacattat ccacttgttt actctgacca acttgggcgc gcctgcagcc
     8101 ttcaagtact tcgacaccac catagacaga aagcggtaca cctctacaaa ggaggtcctg
     8161 gacgccacac tgattcatca gtcaattacg gggctctatg aaacaagaat cgacctctct
     8221 cagctcggtg gagacagcag ggctgacccc aagaagaaga ggaaggtggc tagcgatgct
     8281 aagtcactga ctgcctggtc ccggacactg gtgaccttca aggatgtgtt tgtggacttc
     8341 accagggagg agtggaagct gctggacact gctcagcaga tcctgtacag aaatgtgatg
     8401 ctggagaact ataagaacct ggtttccttg ggttatcagc ttactaagcc agatgtgatc
     8461 ctccggttgg agaagggaga agagccctgg ctggtggaga gagaaattca ccaagagacc
     8521 catcctgatt cagagactgc atttgaaatc aaatcatcag ttccgaaaaa gaaacgcaaa
     8581 gttgctagcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc
     8641 cctggtaccg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     8701 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     8761 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     8821 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     8881 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     8941 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     9001 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     9061 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     9121 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     9181 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     9241 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     9301 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     9361 aagaccggtt gataatagat aacttcgtat agcatacatt atacgaagtt atgaattcga
     9421 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     9481 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     9541 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     9601 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     9661 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     9721 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     9781 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     9841 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     9901 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     9961 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
    10021 cccgcatcga taccgtcgac ctcgagacct agaaaaacat ggagcaatca caagtagcaa
    10081 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
    10141 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
    10201 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
    10261 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta
    10321 cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca agctagtacc
    10381 agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct tgttacaccc
    10441 tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga ggtttgacag
    10501 ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg ggtctctctg
    10561 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
    10621 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
    10681 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt
    10741 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
    10801 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
    10861 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
    10921 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
    10981 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
    11041 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
    11101 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
    11161 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
    11221 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
    11281 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
    11341 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
    11401 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
    11461 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
    11521 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
    11581 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
    11641 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
    11701 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
    11761 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
    11821 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
    11881 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
    11941 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
    12001 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
    12061 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
    12121 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
    12181 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
    12241 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
    12301 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
    12361 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
    12421 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
    12481 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
    12541 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
    12601 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
    12661 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
    12721 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
    12781 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
    12841 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
    12901 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
    12961 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
    13021 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
    13081 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
    13141 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
    13201 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
    13261 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
    13321 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
    13381 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
    13441 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
    13501 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
    13561 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
    13621 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
    13681 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
    13741 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
    13801 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
    13861 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
    13921 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
    13981 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
    14041 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
    14101 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
    14161 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
    14221 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
    14281 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
    14341 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
    14401 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
    14461 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
    14521 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
    14581 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
    14641 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    14701 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
    14761 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
    14821 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
    14881 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
    14941 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tt
//
