LOCUS       sequence-334558-       13702 bp    DNA     circular SYN 14-APR-2025
DEFINITION  PGK+ circuit on an episomal vector. This plasmid encodes eGFP to 
            report circuit output levels. This plasmid also encodes a separate 
            mCherry expression cassette to monitor plasmid dosage..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13702)
  AUTHORS   Yang J, Lee J, Land MA, Lai S, Igoshin OA, St-Pierre F
  TITLE     A synthetic circuit for buffering gene dosage variation between 
            individual mammalian cells.
  JOURNAL   Nat Commun. 2021 Jul 5;12(1):4132. doi: 10.1038/s41467-021-23889-0.
  PUBMED    34226556
REFERENCE   2  (bases 1 to 13702)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2021-07-5"; volume: "12(1):4132. doi"; pages: " 
            10.1038/s41467-021-23889-0"
COMMENT     Sequence Label: PGK+ episome
FEATURES             Location/Qualifiers
     source          1..13702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      667..1255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1156..1175
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1686..1831
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             1874..2911
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     primer_bind     complement(2909..2928)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2953..3001
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        3545..4044
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3580..3603)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3977..3996
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    4051..4069
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4077..4095
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          4278..4850
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      4862..4871
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4868..4870,4871..4873,4874..5587)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4868..4870,4871..4873,4874..5584)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4913..4934)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5174..5193)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5521..5542
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5607..5741
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5631..5650)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5685..5704
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5884..5906)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        6155..7333
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6386..7324
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7281..7301
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      7372..7381
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7378..8088
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(7523..7541)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7791..7810
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(8180..8199)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8245..8300
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8299..8318)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     CDS             complement(10787..12712)
                     /codon_start=1
                     /product="Epstein-Barr nuclear antigen 1, also known as 
                     EBNA-1"
                     /label=EBNA1
                     /note="crucial for latent viral infection, and for episomal
                     amplification of vectors containing the oriP origin (Okita 
                     et al., 2013)"
                     /translation="MSDEGPGTGPGNGLGEKGDTSGPEGSGGSGPQRRGGDNHGRGRGR
                     GRGRGGGRPGAPGGSGSGPRHRDGVRRPQKRPSCIGCKGTHGGTGAGAGAGGAGAGGAG
                     AGGGAGAGGGAGGAGGAGGAGXGGGAGXGGGXGGXGGXGXGGGXGXGGGXGGXGXGGGX
                     GGXGGXGXGGGXGXGGGAGGAGAGGGXGGAGGAGAGGGXGXGGAGGXGGXGAGGAGXGG
                     GAGGAGGAGAGGAGAGGAGAGGAGAGGAGGAGAGGAGGAGXGGXGGAGXGGGAGGXGAG
                     GGXGGXGXGGXGGXGXGGXGGXGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRG
                     GSGGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPP
                     PGRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGR
                     RKKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSL
                     YNLRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLK
                     DAIKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDE
                     GEEGQE"
     primer_bind     complement(13147..13165)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        13233..13337
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(13338..13406,13407..13702,1..496)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(13556..13575)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
       61 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
      121 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
      181 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
      241 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
      301 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
      361 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
      421 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
      481 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
      541 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
      601 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
      661 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      721 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      781 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
      841 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      901 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      961 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1021 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     1081 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1141 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1201 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1261 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttgaagct gtccctgatg
     1321 gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga
     1381 agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac
     1441 gtagcccagc gcgtcggccc cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg
     1501 atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg
     1561 gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccctgcttc atccccgtgg
     1621 cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat atatttgcat gtctttagtt
     1681 ctatgatgac acaaaccccg cccagcgtct tgtcattggc gaattcgaac acgcagatgc
     1741 agtcggggcg gcgcggtccg aggtccactt cgcatattaa ggtgacgcgt gtggcctcga
     1801 acaccgagcg accctgcagc gacccgctta acagcgtcaa cagcgtgccg cagatcccgg
     1861 ggggcaatga gatatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     1921 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     1981 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     2041 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     2101 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     2161 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     2221 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     2281 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     2341 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     2401 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     2461 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     2521 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     2581 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     2641 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     2701 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     2761 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     2821 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     2881 tccggatcgg gagatggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     2941 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     3001 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     3061 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3121 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagccac tggccccgtg
     3181 ggttagggac ggggtccccc atggggaatg gtttatggtt cgtgggggtt attattttgg
     3241 gcgttgcgtg gggtcaggtc cacgactgga ctgagcagac agacccatgg tttttggatg
     3301 gcctgggcat ggaccgcatg tactggcgcg acacgaacac cgggcgtctg tggctgccaa
     3361 acacccccga cccccaaaaa ccaccgcgcg gatttctggc gtgccaagct agtcgaccaa
     3421 ttctcatgtt tgacagctta tcatcgcaga tccgggcaac gttgttgcca ttgctgcagg
     3481 cgcagaactg gtaggtatgg aagatctccc gatcccctat ggtgcactct cagtacaagc
     3541 tagcgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     3601 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     3661 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     3721 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     3781 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     3841 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     3901 aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg
     3961 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
     4021 tcttcctcat ctccgggcct ttcgcgcgcg tccctatcag tgatagagat cagatctccc
     4081 tatcagtgat agagagctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
     4141 ggactctagc gtttaaactt aagctgggta cccggggatc ctctagggcc tctgagctat
     4201 tccagaagta gtgaagaggc ttttttggag gcctaggctt ttgcaaaaag ctccggatcg
     4261 atcctgagaa cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg
     4321 taaaattcat gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat
     4381 gtcccttgta tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt
     4441 gacaaccatt gtctcctctt attttctttt cattttctgt aactttttcg ttaaacttta
     4501 gcttgcattt gtaacgaatt tttaaattca cttttgttta tttgtcagat tgtaagtact
     4561 ttctctaatc actttttttt caaggcaatc agggtatatt atattgtact tcagcacagt
     4621 tttagagaac aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg
     4681 gctggcgtgg aaatattctt attggtagaa acaactacat cctggtcatc atcctgcctt
     4741 tctctttatg gttacaatga tatacactgt ttgagatgag gataaaatac tctgagtcca
     4801 aaccgggccc ctctgctaac catgttcatg ccttcttctt tttcctacag gtcctgcagg
     4861 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     4921 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     4981 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     5041 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     5101 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     5161 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     5221 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     5281 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     5341 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     5401 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     5461 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     5521 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     5581 caagtagctc gaggccggca aggccggatc cagacatgat aagatacatt gatgagtttg
     5641 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     5701 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     5761 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     5821 acaaatgtgg tatggctgat tatgatccgg ctgcctcgcg cgtttcggtg atgacggtga
     5881 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     5941 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     6001 gaggtcgact ctagaggatc gatccccgcc ccggacgaac taaacctgac tacgacatct
     6061 ctgccccttc ttcgcggggc agtgcatgta atcccttcag ttggttggta caacttgcca
     6121 actgggccct gttccacatg aaggagtggg aattggctcc ggtgcccgtc agtgggcaga
     6181 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc
     6241 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     6301 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     6361 caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct
     6421 ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat
     6481 tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg
     6541 agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg
     6601 aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa
     6661 tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca
     6721 agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc
     6781 ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg
     6841 gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc
     6901 gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct
     6961 tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg
     7021 tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc
     7081 cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc
     7141 gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga
     7201 gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga
     7261 gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt
     7321 tcaggtgtcg tgaggtaccg gatcctctag agtcgactcc ggacgtcgac ggccaccatg
     7381 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     7441 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     7501 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     7561 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     7621 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     7681 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     7741 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     7801 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     7861 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     7921 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     7981 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     8041 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaatg agcggccgct
     8101 tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     8161 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     8221 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     8281 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     8341 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     8401 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     8461 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     8521 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     8581 cctgactact cccagtcata gctgtccctc ttctccatgt acctcccagg ggcccaggaa
     8641 gactacggga ggctacacca acgtcaatca gaggggcctg tgtagctacc gataagcgga
     8701 ccctcaagag ggcattagca atagtgttta taaggccccc ttgttaaccc taaacgggta
     8761 gcatatgctt cccgggtagt agtatatact atccagacta accctaattc aatagcatat
     8821 gttacccaac gggaagcata tgctatcgaa ttagggttag taaaagggtc ctaaggaaca
     8881 gcgatatctc ccaccccatg agctgtcacg gttttattta catggggtca ggattccacg
     8941 agggtagtga accattttag tcacaagggc agtggctgaa gatcaaggag cgggcagtga
     9001 actctcctga atcttcgcct gcttcttcat tctccttcgt ttagctaata gaataactgc
     9061 tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa caaggtttca ggtgacgccc
     9121 ccagaataaa atttggacgg ggggttcagt ggtggcattg tgctatgaca ccaatataac
     9181 cctcacaaac cccttgggca ataaatacta gtgtaggaat gaaacattct gaatatcttt
     9241 aacaatagaa atccatgggg tggggacaag ccgtaaagac tggatgtcca tctcacacga
     9301 atttatggct atgggcaaca cataatccta gtgcaatatg atactggggt tattaagatg
     9361 tgtcccaggc agggaccaag acaggtgaac catgttgtta cactctattt gtaacaaggg
     9421 gaaagagagt ggacgccgac agcagcggac tccactggtt gtctctaaca cccccgaaaa
     9481 ttaaacgggg ctccacgcca atggggccca taaacaaaga caagtggcca ctcttttttt
     9541 tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc cgccctgcgg ttttggactg
     9601 taaaataagg gtgtaataac ttggctgatt gtaaccccgc taaccactgc ggtcaaacca
     9661 cttgcccaca aaaccactaa tggcaccccg gggaatacct gcataagtag gtgggcgggc
     9721 caagataggg gcgcgattgc tgcgatctgg aggacaaatt acacacactt gcgcctgagc
     9781 gccaagcaca gggttgttgg tcctcatatt cacgaggtcg ctgagagcac ggtgggctaa
     9841 tgttgccatg ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta
     9901 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     9961 tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta
    10021 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
    10081 tatccgggta gcatatgcta tcctcatgca tatacygtta gcatatgcta tccttcaaat
    10141 atctggatag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
    10201 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
    10261 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
    10321 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctcatgcat
    10381 atacagtcag catatgatac ccagtagtag agtgggagtg ctatcctttg catatgccgc
    10441 cacctcccaa gggggcgtga attttcgctg cttgtccttt tcctgctggt tgctcccatt
    10501 cttaggtgaa tttaaggagg ccaggctaaa gccgtcgcat gtctgattgc tcaccaggta
    10561 aatgtcgcta atgttttcca acgcgagaag gtgttgagcg cggagctgag tgacgtgaca
    10621 acatgggtat gcccaattgc cccatgttgg gaggacgaaa atggtgacaa gacagatggc
    10681 cagaaataca ccaacagcac gcatgatgtc tactggggat ttattcttta gtgcggggga
    10741 atacacggct tttaatacga ttgagggcgt ctcctaacaa gttacatcac tcctgccctt
    10801 cctcaccctc atctccatca cctccttcat ctccgtcatc tccgtcatca ccctccgcgg
    10861 cagccccttc caccataggt ggaaaccagg gaggcaaatc tactccatcg tcaaagctgc
    10921 acacagtcac cctgatattg caggtaggag cgggctttgt cataacaagg tccttaatcg
    10981 catccttcaa aacctcagca aatatatgag tttgtaaaaa gaccatgaaa taacagacaa
    11041 tggactccct tagcgggcca ggttgtgggc cgggtccagg ggccattcca aaggggagac
    11101 gactcaatgg tgtaagacga cattgtggaa tagcaagggc agttcctcgc cttaggttgt
    11161 aaagggaggt cttactacct ccatatacga acacaccggc gacccaagtt ccttcgtcgg
    11221 tagtcctttc tacgtgactc ctagccagga gagctcttaa accttctgca atgttctcaa
    11281 atttcgggtt ggaacctcct tgaccacgat gctttccaaa ccaccctcct tttttgcgcc
    11341 tgcctccatc accctgaccc cggggtccag tgcttgggcc ttctcctggg tcatctgcgg
    11401 ggccctgctc tatcgctccc gggggcacgt caggctcacc atctgggcca ccttcttggt
    11461 ggtattcaaa ataatcggct tcccctacag ggtggaaaaa tggccttcta cctggagggg
    11521 gcctgcgcgg tggagacccg gatgatgatg actgactact gggactcctg ggcctctttt
    11581 ctccacgtcc acgacctctc cccctggctc tttcacgact tccccccctg gctctttcac
    11641 gtcctctacc ccggcggcct ccactacctc ctcgaccccg gcctccacta cctcctcgac
    11701 cccggcctcc actgcctcct cgaccccggc ctccacctcc tgctcctgcc cctcctgctc
    11761 ctgcccctcc tcctgctcct gcccctcctg cccctcctgc tcctscccct cctscccctc
    11821 ctsctcctsc ccctcctscc cctcctsctc ctscccctcc tscccctcct cctgctccts
    11881 cccctcctgc ccctcctcct sctcctgccc ctcctscccc tcctsctcct gcccctcctg
    11941 cccctcctgc tcctgcccct cctgcccctc ctgctcctgc ccctcctgct cctgcccctc
    12001 ctgctcctgc ccctcctgct cctgcccctc ctgcccctcc tgcccctcct cctsctcctg
    12061 cccctcctgc tcctscccct cctscccctc ctgcccctcc tsctcctscc cctcctcctg
    12121 ctcctgcccc tcctgcccct cctscccctc ctcctgctcc tgcccctcct gcccctcctc
    12181 ctsctcctsc ccctcctcct sctcctsccc ctcctscccc tcctscccct cctcctsctc
    12241 ctscccctcc tscccctcct cctsctccts cccctcctcc tsctcctscc cctcctsccc
    12301 ctcctscccc tcctcctsct cctgcccctc ctcctsctcc tgcccctcct gcccctcctg
    12361 cccctcctgc ccctcctcct gctcctgccc ctcctcctgc tcctgcccct cctgctcctg
    12421 cccctcccgc tcctgctcct gctcctgttc caccgtgggt ccctttgcag ccaatgcaac
    12481 ttggacgttt ttggggtctc cggacaccat ctctatgtct tggccctgat cctgagccgc
    12541 ccggggctcc tggtcttccg cctcctcgtc ctcgtcctct tccccgtcct cgtccatggt
    12601 tatcaccccc tcttctttga ggtccactgc cgccggagcc ttctggtcca gatgtgtctc
    12661 ccttctctcc taggccattt ccaggtcctg tacctggccc ctcgtcagac atgattcaca
    12721 ctaaaagaga tcaatagaca tctttattag acgacgctca gtgaatacag ggagtgcaga
    12781 ctcctgcccc ctccaacagc ccccccaccc tcatcccctt catggtcgct gtcagacaga
    12841 tccaggtctg aaaattcccc atcctccgaa ccatcctcgt cctcatcacc aattactcgc
    12901 agcccggaaa actcccgctg aacatcctca agatttgcgt cctgagcctc aagccaggcc
    12961 tcaaattcct cgtccccctt tttgctggac ggtagggatg gggattctcg ggacccctcc
    13021 tcttcctctt caaggtcacc agacagagat gctactgggg caacggaaga aaagctgggt
    13081 gcggcctgtg aggatcagct tatcgatgat aagctgtcaa acatgagaat tcttgaagac
    13141 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
    13201 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
    13261 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
    13321 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
    13381 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
    13441 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
    13501 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
    13561 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
    13621 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
    13681 tgacagtaag agaattatgc ag
//
