LOCUS       sequence-334220-       13937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV+ 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 13937)
  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 13937)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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: CMV+ episome
FEATURES             Location/Qualifiers
     source          1..13937
                     /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)"
     primer_bind     complement(3534..3553)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3725..4104
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4105..4308
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4259..4279
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4304..4322
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4330..4348
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          4531..5103
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      5115..5124
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5121..5123,5124..5126,5127..5840)
                     /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(5121..5123,5124..5126,5127..5837)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5166..5187)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5427..5446)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5774..5795
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5860..5994
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5884..5903)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5938..5957
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6137..6159)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        6408..7586
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6639..7577
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7534..7554
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      7625..7634
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7631..8341
                     /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(7776..7794)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8044..8063
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(8433..8452)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8498..8553
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8552..8571)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     CDS             complement(11022..12947)
                     /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
                     AGGGAGAGGGAGGAGGAGGXGXGGGAGXGGGXGGXGGXGXGGGXGXGGGXGGAGXGGGX
                     GGXGGAGAGGGAGAGGGAGGAGAGGGXGGAGGXGXGGGXGAGGAGGAGGXGAGGAGAGG
                     GAGGAGGAGAGGAGAGGAGAGGAGXGGAGGAGAGGXGGXGAGGAGGAGAGGGAGGAGAG
                     GGAGGXGAGGXGGAGAGGXGGAGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRG
                     GSGGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPP
                     PGRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGR
                     RKKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSL
                     YNLRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLK
                     DAIKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDE
                     GEEGQE"
     primer_bind     complement(13382..13400)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        13468..13572
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(13573..13641,13642..13937,1..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(13791..13810)
                     /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 cagtacaatc
     3541 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     3601 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     3661 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     3721 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     3781 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     3841 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     3901 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     3961 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     4021 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     4081 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     4141 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     4201 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     4261 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcatccctat cagtgataga
     4321 gatcagatct ccctatcagt gatagagagc tgttttgacc tccatagaag acaccgggac
     4381 cgatccagcc tccggactct agcgtttaaa cttaagctgg gtacccgggg atcctctagg
     4441 gcctctgagc tattccagaa gtagtgaaga ggcttttttg gaggcctagg cttttgcaaa
     4501 aagctccgga tcgatcctga gaacttcagg gtgagtttgg ggacccttga ttgttctttc
     4561 tttttcgcta ttgtaaaatt catgttatat ggagggggca aagttttcag ggtgttgttt
     4621 agaatgggaa gatgtccctt gtatcaccat ggaccctcat gataattttg tttctttcac
     4681 tttctactct gttgacaacc attgtctcct cttattttct tttcattttc tgtaactttt
     4741 tcgttaaact ttagcttgca tttgtaacga atttttaaat tcacttttgt ttatttgtca
     4801 gattgtaagt actttctcta atcacttttt tttcaaggca atcagggtat attatattgt
     4861 acttcagcac agttttagag aacaattgtt ataattaaat gataaggtag aatatttctg
     4921 catataaatt ctggctggcg tggaaatatt cttattggta gaaacaacta catcctggtc
     4981 atcatcctgc ctttctcttt atggttacaa tgatatacac tgtttgagat gaggataaaa
     5041 tactctgagt ccaaaccggg cccctctgct aaccatgttc atgccttctt ctttttccta
     5101 caggtcctgc aggcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     5161 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     5221 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     5281 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     5341 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     5401 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     5461 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     5521 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     5581 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     5641 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     5701 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     5761 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     5821 tggacgagct gtacaagtag ctcgaggccg gcaaggccgg atccagacat gataagatac
     5881 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     5941 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     6001 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     6061 aagtaaaacc tctacaaatg tggtatggct gattatgatc cggctgcctc gcgcgtttcg
     6121 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     6181 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     6241 ggggcgcagc catgaggtcg actctagagg atcgatcccc gccccggacg aactaaacct
     6301 gactacgaca tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg
     6361 gtacaacttg ccaactgggc cctgttccac atgaaggagt gggaattggc tccggtgccc
     6421 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     6481 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     6541 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     6601 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
     6661 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
     6721 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
     6781 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
     6841 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     6901 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     6961 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     7021 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     7081 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     7141 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     7201 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
     7261 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     7321 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     7381 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     7441 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     7501 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     7561 tttttcttcc atttcaggtg tcgtgaggta ccggatcctc tagagtcgac tccggacgtc
     7621 gacggccacc atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
     7681 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     7741 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     7801 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     7861 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     7921 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     7981 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     8041 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     8101 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     8161 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     8221 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     8281 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta
     8341 atgagcggcc gcttcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     8401 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
     8461 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     8521 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     8581 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     8641 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     8701 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     8761 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     8821 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctcca tgtacctccc
     8881 aggggcccag gaagactacg ggaggctaca ccaacgtcaa tcagaggggc ctgtgtagct
     8941 accgataagc ggaccctcaa gagggcatta gcaatagtgt ttataaggcc cccttgttaa
     9001 ccctaaacgg gtagcatatg cttcccgggt agtagtatat actatccaga ctaaccctaa
     9061 ttcaatagca tatgttaccc aacgggaagc atatgctatc gaattagggt tagtaaaagg
     9121 gtcctaagga acagcgatat ctcccacccc atgagctgtc acggttttat ttacatgggg
     9181 tcaggattcc acgagggtag tgaaccattt tagtcacaag ggcagtggct gaagatcaag
     9241 gagcgggcag tgaactctcc tgaatcttcg cctgcttctt cattctcctt cgtttagcta
     9301 atagaataac tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga aaacaaggtt
     9361 tcaggtgacg cccccagaat aaaatttgga cggggggttc agtggtggca ttgtgctatg
     9421 acaccaatat aaccctcaca aaccccttgg gcaataaata ctagtgtagg aatgaaacat
     9481 tctgaatatc tttaacaata gaaatccatg gggtggggac aagccgtaaa gactggatgt
     9541 ccatctcaca cgaatttatg gctatgggca acacataatc ctagtgcaat atgatactgg
     9601 ggttattaag atgtgtccca ggcagggacc aagacaggtg aaccatgttg ttacactcta
     9661 tttgtaacaa ggggaaagag agtggacgcc gacagcagcg gactccactg gttgtctcta
     9721 acacccccga aaattaaacg gggctccacg ccaatggggc ccataaacaa agacaagtgg
     9781 ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca cgccgccctg
     9841 cggttttgga ctgtaaaata agggtgtaat aacttggctg attgtaaccc cgctaaccac
     9901 tgcggtcaaa ccacttgccc acaaaaccac taatggcacc ccggggaata cctgcataag
     9961 taggtgggcg ggccaagata ggggcgcgat tgctgcgatc tggaggacaa attacacaca
    10021 cttgcgcctg agcgccaagc acagggttgt tggtcctcat attcacgagg tcgctgagag
    10081 cacggtgggc taatgttgcc atgggtagca tatactaccc aaatatctgg atagcatatg
    10141 ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
    10201 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg
    10261 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
    10321 ctatcctaat ctgtatccgg gtagcatatg ctatcctcat gcatataccc caaatatctg
    10381 gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
    10441 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
    10501 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
    10561 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctca tgcatataca
    10621 gtcagcatat gatacccagt agtagagtgg gagtgctatc ctttgcatat gccgccacct
    10681 cccaaggggg cgtgaatttt cgctgcttgt ccttttcctg ctggttgctc ccattcttag
    10741 gtgaatttaa ggaggccagg ctaaagccgt cgcatgtctg attgctcacc aggtaaatgt
    10801 cgctaatgtt ttccaacgcg agaaggtgtt gagcgcggag ctgagtgacg tgacaacatg
    10861 ggtatgccca attgccccat gttgggagga cgaaaatggt gacaagacag atggccagaa
    10921 atacaccaac agcacgcatg atgtctactg gggatttatt ctttagtgcg ggggaataca
    10981 cggcttttaa tacgattgag ggcgtctcct aacaagttac atcactcctg cccttcctca
    11041 ccctcatctc catcacctcc ttcatctccg tcatctccgt catcaccctc cgcggcagcc
    11101 ccttccacca taggtggaaa ccagggaggc aaatctactc catcgtcaaa gctgcacaca
    11161 gtcaccctga tattgcaggt aggagcgggc tttgtcataa caaggtcctt aatcgcatcc
    11221 ttcaaaacct cagcaaatat atgagtttgt aaaaagacca tgaaataaca gacaatggac
    11281 tcccttagcg ggccaggttg tgggccgggt ccaggggcca ttccaaaggg gagacgactc
    11341 aatggtgtaa gacgacattg tggaatagca agggcagttc ctcgccttag gttgtaaagg
    11401 gaggtcttac tacctccata tacgaacaca ccggcgaccc aagttccttc gtcggtagtc
    11461 ctttctacgt gactcctagc caggagagct cttaaacctt ctgcaatgtt ctcaaatttc
    11521 gggttggaac ctccttgacc acgatgcttt ccaaaccacc ctcctttttt gcgcctgcct
    11581 ccatcaccct gaccccgggg tccagtgctt gggccttctc ctgggtcatc tgcggggccc
    11641 tgctctatcg ctcccggggg cacgtcaggc tcaccatctg ggccaccttc ttggtggtat
    11701 tcaaaataat cggcttcccc tacagggtgg aaaaatggcc ttctacctgg agggggcctg
    11761 cgcggtggag acccggatga tgatgactga ctactgggac tcctgggcct cttttctcca
    11821 cgtccacgac ctctccccct ggctctttca cgacttcccc ccctggctct ttcacgtcct
    11881 ctaccccggc ggcctccact acctcctcga ccccggcctc cactacctcc tcgaccccgg
    11941 cctccactgc ctcctcgacc ccggcctcca cctcctgctc ctgcccctcc tgctcctgcc
    12001 cctcctcctg ctcctgcccc tcctgcccct cctgctcctg cccctcctsc ccctcctgct
    12061 cctgcccctc ctscccctcc tgctcctscc cctcctgccc ctcctcctgc tcctgcccct
    12121 cctgcccctc ctcctgctcc tgcccctcct gcccctcctg ctcctscccc tcctscccct
    12181 cctgctcctg cccctcctgc ccctcctsct cctgcccctc ctgctcctgc ccctcctgct
    12241 cctgcccctc ctgctcctgc ccctcctgcc cctcctgccc ctcctcctgc tcctgcccct
    12301 cctgctccts cccctcctgc ccctcctgcc cctcctgctc ctscccctcc tcctsctcct
    12361 scccctcctg cccctcctsc ccctcctcct gctcctgccc ctcctgcccc tcctcctgct
    12421 cctgcccctc ctcctgctcc tgcccctcct scccctccts cccctcctcc tsctcctgcc
    12481 cctcctsccc ctcctcctsc tcctscccct cctcctsctc ctscccctcc tscccctcct
    12541 scccctcctc ctsctcctgc ccctcctcct sctcctsccc ctcctgcccc tcctgcccct
    12601 cctgcccctc ctcctgctcc tgcccctcct cctgctcctg cccctcctgc tcctgcccct
    12661 cccgctcctg ctcctgctcc tgttccaccg tgggtccctt tgcagccaat gcaacttgga
    12721 cgtttttggg gtctccggac accatctcta tgtcttggcc ctgatcctga gccgcccggg
    12781 gctcctggtc ttccgcctcc tcgtcctcgt cctcttcccc gtcctcgtcc atggttatca
    12841 ccccctcttc tttgaggtcc actgccgccg gagccttctg gtccagatgt gtctcccttc
    12901 tctcctaggc catttccagg tcctgtacct ggcccctcgt cagacatgat tcacactaaa
    12961 agagatcaat agacatcttt attagacgac gctcagtgaa tacagggagt gcagactcct
    13021 gccccctcca acagcccccc caccctcatc cccttcatgg tcgctgtcag acagatccag
    13081 gtctgaaaat tccccatcct ccgaaccatc ctcgtcctca tcaccaatta ctcgcagccc
    13141 ggaaaactcc cgctgaacat cctcaagatt tgcgtcctga gcctcaagcc aggcctcaaa
    13201 ttcctcgtcc ccctttttgc tggacggtag ggatggggat tctcgggacc cctcctcttc
    13261 ctcttcaagg tcaccagaca gagatgctac tggggcaacg gaagaaaagc tgggtgcggc
    13321 ctgtgaggat cagcttatcg atgataagct gtcaaacatg agaattcttg aagacgaaag
    13381 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
    13441 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
    13501 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
    13561 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
    13621 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
    13681 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
    13741 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
    13801 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
    13861 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
    13921 gtaagagaat tatgcag
//
