LOCUS       sequence-334453-        9257 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PGK+ plasmid that also encodes separate mCherry cassette. mCherry is
            used to monitor plasmid dosage..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9257)
  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 9257)
  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: PGK+ plasmid with onboard mCherry cassette
FEATURES             Location/Qualifiers
     source          1..9257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    594..715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(631..650)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     685..704
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(764..780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(764..780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(777..799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    788..804
                     /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(join(812..818,819..836,837..842))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    857..878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(995..1012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1125..2303
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1356..2294
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2251..2271
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      2342..2351
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2348..3058
                     /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(2493..2511)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2761..2780
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3150..3169)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3215..3270
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3269..3288)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(3647..4235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3727..3746)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4406..5197,5198..5266))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5029..5048
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5267..5371)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5446..5465)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5653..6152
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5688..5711)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6085..6104
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    6159..6177
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6185..6203
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          6386..6958
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      6970..6979
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6976..6978,6979..6981,6982..7695)
                     /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(6976..6978,6979..6981,6982..7692)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7021..7042)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7282..7301)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7629..7650
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7757..7777)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(8130..8147)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8136..8360
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8493..8512)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    8644..8691
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             join(8699..9257,1..464)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             join(8699..9257,1..464)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca
       61 atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg
      121 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct
      181 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
      241 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
      301 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
      361 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
      421 gtggaaaccg acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt
      481 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
      541 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
      601 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
      661 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
      721 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
      781 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
      841 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
      901 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
      961 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1021 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1081 aatcagggga taacgcagga aagaacatgt aaggagtggg aattggctcc ggtgcccgtc
     1141 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     1201 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     1261 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     1321 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg
     1381 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc
     1441 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt
     1501 gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcccggg cgctggggcc
     1561 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag
     1621 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa
     1681 atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg
     1741 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa
     1801 tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt
     1861 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa
     1921 gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag
     1981 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt
     2041 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt
     2101 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg
     2161 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg
     2221 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt
     2281 ttcttccatt tcaggtgtcg tgaggtaccg gatcctctag agtcgactcc ggacgtcgac
     2341 ggccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     2401 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     2461 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     2521 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     2581 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     2641 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     2701 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     2761 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     2821 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     2881 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     2941 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     3001 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaatg
     3061 agcggccgct tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
     3121 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac
     3181 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     3241 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     3301 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
     3361 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     3421 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     3481 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     3541 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcacatg tgagcaaaag
     3601 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3661 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3721 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3781 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3841 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3901 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3961 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4021 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4081 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4141 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4201 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4261 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4321 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4381 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4441 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4501 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4561 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4621 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4681 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4741 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4801 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4861 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4921 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4981 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5041 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5101 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5161 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5221 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5281 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5341 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5401 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     5461 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     5521 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     5581 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtgacgg
     5641 atcgggagat ctgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag
     5701 cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat
     5761 ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc
     5821 ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa
     5881 tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc
     5941 gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga
     6001 ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg
     6061 cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc
     6121 tgttctcctc ttcctcatct ccgggccttt cgcgcgcgtc cctatcagtg atagagatca
     6181 gatctcccta tcagtgatag agagctgttt tgacctccat agaagacacc gggaccgatc
     6241 cagcctccgg actctagcgt ttaaacttaa gctgggtacc cggggatcct ctagggcctc
     6301 tgagctattc cagaagtagt gaagaggctt ttttggaggc ctaggctttt gcaaaaagct
     6361 ccggatcgat cctgagaact tcagggtgag tttggggacc cttgattgtt ctttcttttt
     6421 cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt tgtttagaat
     6481 gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct ttcactttct
     6541 actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa ctttttcgtt
     6601 aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt tgtcagattg
     6661 taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat attgtacttc
     6721 agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat ttctgcatat
     6781 aaattctggc tggcgtggaa atattcttat tggtagaaac aactacatcc tggtcatcat
     6841 cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga taaaatactc
     6901 tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctttt tcctacaggt
     6961 cctgcaggcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     7021 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     7081 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     7141 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     7201 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     7261 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     7321 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     7381 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     7441 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     7501 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     7561 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     7621 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     7681 gagctgtaca agtaggcggc cgcaatcaac ctctggatta caaaatttgt gaaagattga
     7741 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     7801 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     7861 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     7921 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     7981 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     8041 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcctcgagtc
     8101 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     8161 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     8221 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     8281 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     8341 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     8401 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     8461 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     8521 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     8581 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac
     8641 ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa
     8701 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     8761 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     8821 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     8881 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     8941 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     9001 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     9061 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     9121 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     9181 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     9241 gctttgggcc gaggact
//
