LOCUS       40924_46598       13274 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWM91-L6-PT3-T4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13274)
  AUTHORS   Lee JW, Gyorgy A, Cameron DE, Pyenson N, Choi KR, Way JC, Silver PA,
            Del Vecchio D, Collins JJ.
  TITLE     Creating Single-Copy Genetic Circuits
  JOURNAL   Mol. Cell 63 (2), 329-336 (2016)
  PUBMED    27425413
REFERENCE   2  (bases 1 to 13274)
  AUTHORS   Lee JW, Gyorgy A, Cameron DE, Pyenson N, Choi KR, Way JC, Silver PA,
            Del Vecchio D, Collins JJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAY-2016) Biological Engineering, MIT, 45 Carleton St,
            MIT Building E25-Rm302, Cambridge, MA 02142, USA
REFERENCE   3  (bases 1 to 13274)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 13274)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Cell";
            date: "2016"; volume: "63"; issue: "2"; pages: "329-336"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-MAY-2016) Biological Engineering, MIT, 45 Carleton St, MIT 
            Building E25-Rm302, Cambridge, MA 02142, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..13274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(60..515)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     657..673
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        683..701
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     727..743
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    776..1194
                     /label=similar to lacZ (upstream)
                     /note="similar to lacZ (upstream)"
     terminator      complement(1237..1331)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1394..2107)
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     regulatory      complement(2108..2131)
                     /label=G5
                     /note="G5"
                     /regulatory_class="ribosome_binding_site"
     RBS             2113..2121
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(2149..2222)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     misc_feature    complement(2265..2348)
                     /label=mfLon-specific tag pdt2
                     /note="mfLon-specific tag pdt2"
     CDS             complement(2349..3428)
                     /label=lacI
                     /note="lac repressor"
     RBS             3434..3442
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(3474..3547)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     protein_bind    3566..3585
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    3602..3621
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        3622..3651
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    complement(3658..3677)
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     regulatory      3690..3721
                     /label=T4
                     /note="T4"
                     /regulatory_class="ribosome_binding_site"
     RBS             3708..3716
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             3722..4342
                     /label=TetR
                     /note="tetracycline repressor TetR"
     regulatory      4388..4462
                     /label=Ptrc-2
                     /note="Ptrc-2"
                     /regulatory_class="promoter"
     promoter        4401..4430
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4438..4454
                     /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)."
     prim_transcript 4438
                     /label=transcription start
                     /note="transcription start"
     regulatory      4470..4496
                     /label=M2
                     /note="M2"
                     /regulatory_class="ribosome_binding_site"
     RBS             4483..4491
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             4497..5204
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     terminator      5267..5353
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    5422..5821
                     /label=similar to lacZ (downstream)
                     /note="similar to lacZ (downstream)"
     promoter        complement(5840..5858)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5879..5895)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5903..5919
                     /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(5927..5957)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5972..5993)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6697..7065)
                     /label=traJ
                     /note="oriT-recognizing protein"
     oriT            complement(7098..7207)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        8374..8819
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             8820..10238
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     CDS             complement(11811..12668)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(12669..12773)
                     /label=AmpR promoter
     rep_origin      complement(12883..13271)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 tacgccggac gcatcgtggg gatctggttc cgcgcacatt tccccgaaaa gtgccaccta
       61 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
      121 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
      181 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
      241 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
      301 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
      361 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
      421 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
      481 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
      541 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
      601 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
      661 aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccgggcc
      721 ccccctcgag gtcgacggta tcgataagct tgatatcgaa ttcctgcagc ccgggtttac
      781 gcgccggaga aaaccgcctc gcggtgatgg tgctgcgctg gagtgacggc agttatctgg
      841 aagatcagga tatgtggcgg atgagcggca ttttccgtga cgtctcgttg ctgcataaac
      901 cgactacaca aatcagcgat ttccatgttg ccactcgctt taatgatgat ttcagccgcg
      961 ctgtactgga ggctgaagtt cagatgtgcg gcgagttgcg tgactaccta cgggtaacag
     1021 tttctttatg gcagggtgaa acgcaggtcg ccagcggcac cgcgcctttc ggcggtgaaa
     1081 ttatcgatga gcgtggtggt tatgccgatc gcgtcacact acgtctgaac gtcgaaaacc
     1141 cgaaactgtg gagcgccgaa atcccgaatc tctatcgtgc ggtggttgaa ctgcgcggcc
     1201 gcaaacatga gaattggtct cgacgagtat gcttggattc tcaccaataa aaaacgcccg
     1261 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat
     1321 caacaggagt ccaagcgacg tctgtgcaag tactactgtt ctgcagtcac ttgaattcga
     1381 tacccagctg ttatttgtat agttcatcca tgccatgtgt aatcccagca gcggttacaa
     1441 actcaagaag gaccatgtgg tctctctttt cgttgggatc tttcgaaagg gcagattgtg
     1501 tggacaggta atggttgtct ggtaaaagga cagggccatc gccaatcgga gtattttgtt
     1561 gataatggtc tgctagttga acgcttccat cttcaatgtt gtgtctaatt ttgaagttaa
     1621 ctttgattcc attcttttgt ttgtctgcca tgatgtatac attgtgtgag ttatagttgt
     1681 attccaattt gtgtccaaga atgtttccat cttctttaaa atcaatacct tttaactcga
     1741 ttctattaac aagggtatca ccttcaaact tgacttcagc acgtgtcttg tagttcccgt
     1801 catctttgaa aaatatagtt ctttcctgta cgtaaccttc gggcatggca ctcttgaaaa
     1861 agtcatgctg tttcatgtga tctgggtatc tcgcaaagca ttgaacacca taaccgaaag
     1921 tagtgacaag tgttggccac ggaacaggta gttttccagt agtgcaaata aatttaaggg
     1981 taagttttcc gtatgttgca tcaccttcac cctctccact gacagaaaat ttgtgcccat
     2041 taacatcacc atctaattca acaagaattg ggacaactcc agtgaaaagt tcttctcctt
     2101 tacgcatatt atacctcctt aatttagtgt catctacaac tggcatgcgg tcagtgcgtc
     2161 ctgctgatgt gctcagtata tctatcactg atagggatgt caatctctat cactgatagg
     2221 gaaacgtttc gcagaagctt ccgcaaggta ccactttgcc gcggttattg agacaggtag
     2281 ttcgcctggc ccgcgttcag cataaaggtc ggcacttcgt tggtgttttc ttcgtttttg
     2341 ttcgccgcct gcccgctttc cagtcgggaa acctgtcgtg ccagttgcat taatgaatcg
     2401 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     2461 agtgagacgg gcaacagttg attgcccttc accgcctggc cctgagagag ttgcagcaag
     2521 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     2581 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg
     2641 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     2701 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     2761 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     2821 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     2881 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     2941 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     3001 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     3061 atcagcccac tgacgcgctg cgcgagaaga ttgtgaaccg ccgctttaca ggcttcgacg
     3121 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     3181 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     3241 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     3301 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     3361 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta cagcgttact
     3421 ggtttcatat tgtacctcct taattcttga ttttctatcc tcgtcatgtg agcggtcagt
     3481 gcgtcctgct gatgtgctca gtatatctat cactgatagg gatgtcaatc tctatcactg
     3541 atagggacgg ccgcccgttc catggaattg tgagcgctca caattactag cggccgcatg
     3601 gaattgtgag cgctcacaat tttgacaatt aatcatccgg ctcgtataat gtgtgggaat
     3661 tgtgagcgct cacaatttca cacgtcgacg caccataaag cggaaagtaa ggaggttaga
     3721 tatgtctcgt ttagataaaa gtaaagtgat taacagcgca ttagagctgc ttaatgaggt
     3781 cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag agcagcctac
     3841 attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca ttgagatgtt
     3901 agataggcac catactcact tttgcccttt agaaggggaa agctggcaag attttttacg
     3961 taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag caaaagtaca
     4021 tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat tagccttttt
     4081 atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgctg tggggcattt
     4141 tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag aaagggaaac
     4201 acctactact gatagtatgc cgccattatt acgacaagct atcgaattat ttgatcacca
     4261 aggtgcagag ccagccttct tattcggcct tgaattgatc atctgcggat tagaaaaaca
     4321 acttaaatgt gaaagtgggt cttgagagct cggactgctt aagtcgctcc atatgctcgt
     4381 tcccgggctg aaatgagctg ttgacaatta atcatccggc tcgtataatg tgtggaattg
     4441 tgagcggata acaatttcac accaattgat aatatacaag attaaggagg taaagaatgg
     4501 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggttc
     4561 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     4621 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     4681 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     4741 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     4801 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caagacggcg
     4861 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     4921 agaagaagac tatgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgcgc
     4981 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     5041 tcaagaccac ctacaaggcc aagaagcccg tgcaactgcc cggcgcgtac aacgtcaaca
     5101 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     5161 ccgagggccg ccactccacc ggcggcatgg acgagctgta taagtaagat atctatcgcc
     5221 ctagggaccg tctcgagaga atcaatatta atccaacgcg tggcatcaaa taaaacgaaa
     5281 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     5341 gagtaggaca aatccgccgc cctagactta ggcgttcggc tgcggcgagc ggtatcagct
     5401 cactcaaaga agtggggatc ctttatcagc cggaaaacct accggattga tggtagtggt
     5461 caaatggcga ttaccgttga tgttgaagtg gcgagcgata caccgcatcc ggcgcggatt
     5521 ggcctgaact gccagctggc gcaggtagca gagcgggtaa actggctcgg attagggccg
     5581 caagaaaact atcccgaccg ccttactgcc gcctgttttg accgctggga tctgccattg
     5641 tcagacatgt ataccccgta cgtcttcccg agcgaaaacg gtctgcgctg cgggacgcgc
     5701 gaattgaatt atggcccaca ccagtggcgc ggcgacttcc agttcaacat cagccgctac
     5761 agtcaacagc aactgatgga aaccagccat cgccatctgc tgcacgcgga agaaggcaca
     5821 tgagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     5881 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5941 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6001 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6061 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc agatccagcc gaccaggctt
     6121 tccacgcccg cgtgccgctc catgtcgttc gcgcggttct cggaaacgcg ctgccgcgtt
     6181 tcgtgattgt cacgctcaag cccgtagtcc cgttcgagcg tcgcgcagag gtcagcgagg
     6241 gcgcggtagg cccgatacgg ctcatggatg gtgtttcggg tcgggtgaat cttgttgatg
     6301 gcgatatgga tgtgcaggtt gtcggtgtcg tgatgcacgg cactgacgcg ctgatgctcg
     6361 gcgaagccaa gcccagcgca gatgcggtcc tcaatcgcgc gcaacgtctc cgcgtcgggc
     6421 ttctctcccg cgcggaagct aaccagcagg tgataggtct tgtcggcctc ggaacgggtg
     6481 ttgccgtgct gggtcgccat cacctcggcc atgacagcgg gcagggtgtt tgcctcgcag
     6541 ttcgtgacgc gcacgtgacc caggcgctcg gtcttgcctt gctcgtcggt gatgtacttc
     6601 accagctccg cgaagtcgct cttcttgatg gagcgcatgg ggacgtgctt ggcaatcacg
     6661 cgcacccccc ggccgtttta gcggctaaaa aagtcatggc tctgccctcg ggcggaccac
     6721 gcccatcatg accttgccaa gctcgtcctg cttctcttcg atcttcgcca gcagggcgag
     6781 gatcgtggca tcaccgaacc gcgccgtgcg cgggtcgtcg gtgagccaga gtttcagcag
     6841 gccgcccagg cggcccaggt cgccattgat gcgggccagc tcgcggacgt gctcatagtc
     6901 cacgacgccc gtgattttgt agccctggcc gacggccagc aggtaggccg acaggctcat
     6961 gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg tctggaaggc agtacacctt
     7021 gataggtggg ctgcccttcc tggttggctt ggtttcatca gccatccgct tgccctcatc
     7081 tgttacgccg gcggtagccg gccagcctcg cagagcagga ttcccgttga gcaccgccag
     7141 gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta cttcacctat
     7201 cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc tttggcaaaa
     7261 tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa tgaccccgaa
     7321 gcagggttat gcagcggaaa agcgctgctt ccctgctgtt ttgtggaata tctaccgact
     7381 ggaaacaggc aaatgcagga aattactgaa ctgaggggac aggcgagaga cgatgccaaa
     7441 gagctacacc gacgagctgg ccgagtgggt tgaatcccgc gcggccaaga agcgccggcg
     7501 tgatgaggct gcggttgcgt tcctggcggt gagggcggat gtcgaggcgg cgttagcgtc
     7561 cggctatgcg ctcgtcacca tttgggagca catgcgggaa acggggaagg tcaagttctc
     7621 ctacgagacg ttccgctcgc acgccaggcg gcacatcaag gccaagcccg ccgatgtgcc
     7681 cgcaccgcag gccaaggctg cggaacccgc gccggcaccc aagacgccgg agccacggcg
     7741 gccgaagcag gggggcaagg ctgaaaagcc ggcccccgct gcggccccga ccggcttcac
     7801 cttcaaccca acaccggaca aaaaggatct atgattccct ttgtcaacag caatggatca
     7861 ctgaaaatgg ttcaatgatc acattaagtg gtattcaata ttttcatgaa atgggaattg
     7921 acgttccttc caaacattca cgtaaaatct gttgtgcgtg tttagattgg agtgaacgcc
     7981 gtttccattt aggtgggtac gttggagccg cattattttc gctttatgaa tctaaagggt
     8041 ggttaactcg acatcttggt taccgtgaag ttaccatcac ggaaaaaggt tatgctgctt
     8101 ttaagaccca ctttcacatt taagttgttt ttctaatccg catatgatca attcaaggcc
     8161 gaataagaag gctggctctg caccttggtg atcaaataat tcgatagctt gtcgtaataa
     8221 tggcggcata ctatcagtag taggtgtttc cctttcttct ttagcgactt gatgctcttg
     8281 atcttccaat acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat ataatgcagg
     8341 tcgactctag aggatcgatc ctttttaacc catcacatat acctgccgtt cactattatt
     8401 tagtgaaatg agatattatg atattttctg aattgtgatt aaaaaggcaa ctttatgccc
     8461 atgcaacaga aactataaaa aatacagaga atgaaaagaa acagatagat tttttagttc
     8521 tttaggcccg tagtctgcaa atccttttat gattttctat caaacaaaag aggaaaatag
     8581 accagttgca atccaaacga gagtctaata gaatgaggtc gaaaagtaaa tcgcgcgggt
     8641 ttgttactga taaagcaggc aagacctaaa atgtgtaaag ggcaaagtgt atactttggc
     8701 gtcacccctt acatatttta ggtctttttt tattgtgcgt aactaacttg ccatcttcaa
     8761 acaggagggc tggaagaagc agaccgctaa cacagtacat aaaaaaggag acatgaacga
     8821 tgaacatcaa aaagtttgca aaacaagcaa cagtattaac ctttactacc gcactgctgg
     8881 caggaggcgc aactcaagcg tttgcgaaag aaacgaacca aaagccatat aaggaaacat
     8941 acggcatttc ccatattaca cgccatgata tgctgcaaat ccctgaacag caaaaaaatg
     9001 aaaaatatca agttcctgaa ttcgattcgt ccacaattaa aaatatctct tctgcaaaag
     9061 gcctggacgt ttgggacagc tggccattac aaaacgctga cggcactgtc gcaaactatc
     9121 acggctacca catcgtcttt gcattagccg gagatcctaa aaatgcggat gacacatcga
     9181 tttacatgtt ctatcaaaaa gtcggcgaaa cttctattga cagctggaaa aacgctggcc
     9241 gcgtctttaa agacagcgac aaattcgatg caaatgattc tatcctaaaa gaccaaacac
     9301 aagaatggtc aggttcagcc acatttacat ctgacggaaa aatccgttta ttctacactg
     9361 atttctccgg taaacattac ggcaaacaaa cactgacaac tgcacaagtt aacgtatcag
     9421 catcagacag ctctttgaac atcaacggtg tagaggatta taaatcaatc tttgacggtg
     9481 acggaaaaac gtatcaaaat gtacagcagt tcatcgatga aggcaactac agctcaggcg
     9541 acaaccatac gctgagagat cctcactacg tagaagataa aggccacaaa tacttagtat
     9601 ttgaagcaaa cactggaact gaagatggct accaaggcga agaatcttta tttaacaaag
     9661 catactatgg caaaagcaca tcattcttcc gtcaagaaag tcaaaaactt ctgcaaagcg
     9721 ataaaaaacg cacggctgag ttagcaaacg gcgctctcgg tatgattgag ctaaacgatg
     9781 attacacact gaaaaaagtg atgaaaccgc tgattgcatc taacacagta acagatgaaa
     9841 ttgaacgcgc gaacgtcttt aaaatgaacg gcaaatggta cctgttcact gactcccgcg
     9901 gatcaaaaat gacgattgac ggcattacgt ctaacgatat ttacatgctt ggttatgttt
     9961 ctaattcttt aactggccca tacaagccgc tgaacaaaac tggccttgtg ttaaaaatgg
    10021 atcttgatcc taacgatgta acctttactt actcacactt cgctgtacct caagcgaaag
    10081 gaaacaatgt cgtgattaca agctatatga caaacagagg attctacgca gacaaacaat
    10141 caacgtttgc gccaagcttc ctgctgaaca tcaaaggcaa gaaaacatct gttgtcaaag
    10201 acagcatcct tgaacaagga caattaacag ttaacaaata aaaacgcaaa agaaaatgcc
    10261 gatatcctat tggcattttc ttttatttct tatcaacata aaggtgaatc ccatatgaac
    10321 tatataaaag caggcaaatg gctaaccgta ttcctaacct tttggtaatg actccaactt
    10381 attgatagtg ttttatgttc agataatgcc cgatgacttt gtcatgcagc tccaccgatt
    10441 ttgagaacga cagcgacttc cgtcccagcc gtgccaggtg ctgcctcaga ttcaggttat
    10501 gccgctcaat tcgctgcgta tatcgcttgc tgattacgtg cagctttccc ttcaggcggg
    10561 attcatacag cggccagcca tccgtcatcc atatcaccac gtcaaagggt gacagcaggc
    10621 tcataagacg ccccagcgtc gccatagtgc gttcaccgaa tacgtgcgca acaaccgtct
    10681 tccggagact gtcatacgcg taaaacagcc agcgctggcg cgatttagcc ccgacatagc
    10741 cccactgttc gtccatttcc gcgcagacga tgacgtcact gcccggctgt atgcgcgagg
    10801 ttaccgactg cggcctgagt tttttaagtg acgtaaaatc gtgttgaggc caacgcccat
    10861 aatgcgggct gttgcccggc atccaacgcc attcatggcc atatcaatga ttttctggtg
    10921 cgtaccgggt tgagaagcgg tgtaagtgaa ctgcataaac cagccattga gtaagttttt
    10981 aagcacatca ctatcataag ctttaagttg gttctcttgg atcaatttgc tgacaatggc
    11041 gtttacctta ccagtaatgt attcaaggct aattttttca agttcattcc aaccaatgat
    11101 aggcatcact tcttggatag ggataaggtt tttattatta tcaataatat aatcaagata
    11161 atgttcaaat atactttcta aggcagacca accatttgtt aaatcagttt ttgttgtgat
    11221 gtaggcatca atcataatta attgctgctt ataacaggca ctgagtaatt gttttttatt
    11281 tttaaagtga tgataaaagg cacctttggt caccaacgct tttcccgaga tcctctacgc
    11341 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
    11401 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
    11461 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttggctgc
    11521 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
    11581 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
    11641 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
    11701 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa ttaaaaatga
    11761 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
    11821 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
    11881 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
    11941 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
    12001 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
    12061 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
    12121 gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
    12181 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
    12241 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
    12301 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
    12361 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
    12421 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
    12481 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
    12541 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
    12601 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
    12661 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
    12721 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
    12781 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
    12841 aataggcgta tcacgaggcc ctttcgtctt caagaattcc catgtcagcc gttaagtgtt
    12901 cctgtgtcac tcaaaattgc tttgagaggc tctaagggct tctcagtgcg ttacatccct
    12961 ggcttgttgt ccacaaccgt taaaccttaa aagctttaaa agccttatat attctttttt
    13021 ttcttataaa acttaaaacc ttagaggcta tttaagttgc tgatttatat taattttatt
    13081 gttcaaacat gagagcttag tacgtgaaac atgagagctt agtacgttag ccatgagagc
    13141 ttagtacgtt agccatgagg gtttagttcg ttaaacatga gagcttagta cgttaaacat
    13201 gagagcttag tacgtgaaac atgagagctt agtacgtact atcaacaggt tgaactgctg
    13261 atcttcagat cctc
//
