LOCUS       sequence-419255-       11488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vinculin conformation sensor (VinCS) with vinculin S1033 
            phosphomimetic point mutation (S1033D), in lentiviral expression 
            vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11488)
  AUTHORS   Shoyer TC, Gates EM, Cabe JI, Urs AN, Conway DE, Hoffman BD
  TITLE     Coupling during collective cell migration is controlled by a 
            vinculin mechanochemical switch.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Dec 12;120(50):e2316456120. doi: 
            10.1073/pnas.2316456120. Epub 2023 Dec 6.
  PUBMED    38055737
REFERENCE   2  (bases 1 to 11488)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-12-12"; volume: "120(50):e2316456120. 
            doi"; pages: " 10.1073/pnas.2316456120. Epub 2023 Dec 6"
COMMENT     Sequence Label: pRRL-VinculinCS-S1033D
FEATURES             Location/Qualifiers
     source          1..11488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      82..670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     571..590
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     824..841
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    958..979
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(994..999,1000..1017,1018..1024)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1032..1048
                     /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)."
     primer_bind     1037..1059
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1056..1072
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1056..1072
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1091..1111
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1093..1111
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1139..1365
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1366..1546
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1593..1718
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2211..2444
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2629..2673
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2940..3056
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3116..3495
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3496..3699
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3650..3670
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3744..3762
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6456..7163
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     CDS             join(7728..7730,7731..7733,7734..8447)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7773..7794)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8034..8053)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8381..8402
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    8466..9054
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8519..8539)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8937..8948)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9145..9378
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9450..9571
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9487..9506)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9541..9560
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9611..9746
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(9767..9785)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9795..9812)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9795..9811)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9804..9826)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9953..10408
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10040..10059)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10250..10271
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10434..10538
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10539..10607,10608..11399)
                     /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     complement(10757..10776)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
       61 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
      121 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
      181 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
      241 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
      301 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
      361 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
      421 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
      481 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      541 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      601 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      661 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      721 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      781 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      841 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      901 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      961 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     1021 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     1081 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
     1141 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     1201 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     1261 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     1321 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     1381 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     1441 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     1501 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     1561 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     1621 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     1681 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     1741 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     1801 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     1861 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     1921 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     1981 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     2041 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     2101 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     2161 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     2221 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     2281 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     2341 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     2401 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     2461 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     2521 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     2581 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     2641 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     2701 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     2761 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     2821 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     2881 gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact
     2941 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     3001 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaattcaa aattttatcg
     3061 atggatcccc cgggctcgag ttcgaagatc gatgtacggg ccagatatac gcgttgacat
     3121 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     3181 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     3241 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     3301 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     3361 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     3421 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     3481 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     3541 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     3601 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     3661 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
     3721 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
     3781 cgtttaaact taagcttgcc atgcccgtct tccacacgcg caccatcgag agcatcttgg
     3841 agcccgtggc tcagcagatc tcccacctgg tcatcatgca cgaggagggg gaggtagacg
     3901 gcaaggccat cccggacctc accgcccccg tgtcggccgt gcaggccgct gtcagcaacc
     3961 tggtgcgggt tggaaaagaa actgtgcaga caacagaaga ccagatcttg aaaagggata
     4021 tgccaccagc attcatcaaa gtagagaatg cctgcaccaa gctcgttcga gcagcccaga
     4081 tgctgcaagc agatccttat tcagtaccag ctcgtgacta cctaattgat ggatcaagag
     4141 gcatcctttc tggaacatca gacttacttc tgacatttga tgaagcggag gtccgtaaaa
     4201 tcatccgtgt ctgcaaagga atattggaat atctgactgt ggcagaagta gtagagacta
     4261 tggaggattt ggtgacatat acaaagaatc tagggccagg aatgacaaag atggcgaaaa
     4321 tgatcgatga gagacaacag gaattaactc atcaggaaca tagggttatg ctggtgaact
     4381 ccatgaatac tgtgaaggag ctattgcctg tacttatttc agctatgaag atctttgtaa
     4441 ccacaaaaaa cactaaaagc cagggaatag aagaggcctt gaaaaatcgc aatttcacag
     4501 tagagaaaat gagtgctgag ataaatgaaa taatccgtgt attacaactc acttcctggg
     4561 atgaagatgc ctgggccagc aaggacactg aagccatgaa aagagcttta gccctaatag
     4621 attcaaagat gaaccaggca aaaggctggc tgagagatcc aaacgcacct ccaggggatg
     4681 ctggtgagca agcaataagg cagatccttg atgaagctgg aaaagcagga gaattgtgtg
     4741 caggcaaaga acgcagggag attctgggaa catgcaaaac tttgggccaa atgactgatc
     4801 aacttgctga tctccgagct agaggacagg gtgctacacc gatggcaatg cagaaggcac
     4861 agcaggtgtc acaaggcctg gatttgctca ctgcaaaagt ggagaatgca gcccggaaat
     4921 tggaggccat gacaaactct aagcaggcta ttgcaaagaa gattgatgct gctcagaact
     4981 ggcttgcgga tcccaacggc ggaagtgaag gagaagaaca cattcgagga attatgtctg
     5041 aagcaaggaa agttgcagaa ttgtgtgagg agcctaaaga aagagatgat atccttcgct
     5101 ccttggggga aatctctgct ctgacagcta agctgtcaga tctgcgacga catgggaaag
     5161 gcgactctcc tgaggcccgt gcattggcca agcaaatagc tacatcactt cagaacttac
     5221 agtccaaaac aaacagagct gtagcaaata ctaggccagt taaagctgct gtccatttgg
     5281 agggcaagat tgagcaagcc cagaggtgga tagataatcc tacagttgat gatcggggag
     5341 taggccaggc agcaattcgg ggtttggttg cagaaggtcg tcgtctagcc aatgtcatga
     5401 tgggacctta tcgtcaggac ctgcttgcca aatgtgaccg tgtagaccag ctggctgctc
     5461 agcttgctga ccttgcagca agaggggaag gagagtctcc tcaggctagg gcaattgctg
     5521 ctcagcttca ggactccctg aaggatctca aagcacggat gcaagaagca atgacccagg
     5581 aggtgtctga tgttttcagt gacactacaa cccctattaa gttgttagca gtagcagcca
     5641 ctgctccttc tgatactccc aatagagaag aggtgtttga agaaagagca gcaaattttg
     5701 aaaaccatgc tgctagactg ggagcaacag cagaaaaagc agctgcagtt ggaactgcta
     5761 ataaaactac tgtggaaggc attcaggcaa cagtcaaatc tgcaagggag cttacaccac
     5821 aggtagtatc ggctgctcga atcctcctga gaaatcctgg aaatcaagct gcttatgagc
     5881 attttgagac aatgaaaaac caatggattg ataatgtaga aaagatgaca gggctggtgg
     5941 atgaggccat cgataccaag tctctgttgg atgcatcaga agaggctatt aagaaggatc
     6001 ttgataaatg taaagttgca atggccaata tgcaacctca gatgctggta gctggagcca
     6061 ccagcattgc tagacgagcg aaccgcatcc tgcttgtggc aaaacgggag gttgaaaatt
     6121 cagaagaccc taaattcagg gaggctgtta aagcagcttc tgatgagctg agcaaaacca
     6181 tatcaccgat ggtaatggat gctaaagctg tagcaggaaa tatctctgat cctggtttgc
     6241 agaagagttt cttggattct ggatacagga ttctgggagc tgtggccaaa gtcagagaag
     6301 cctttcagcc tcaggagcca gactttcccc ctcctcctcc tgacctagag catctccatc
     6361 tgactgatga gcttgctcct ccaaaaccac cacttccaga aggtgaggtt cccccaccca
     6421 gaccaccacc ccctgaagaa aaagatgagg tcgagatggt gagcaagggc gaggagacca
     6481 caatgggcgt aatcaagccc gacatgaaga tcaagctgaa gatggagggc aacgtgaatg
     6541 gccacgcctt cgtgatcgag ggcgagggcg agggcaagcc ctacgacggc accaacacca
     6601 tcaacctgga ggtgaaggag ggagcccccc tgcccttctc ctacgacatt ctgaccaccg
     6661 cgttcgccta cggcaacagg gccttcacca agtaccccga cgacatcccc aactacttca
     6721 agcagtcctt ccccgagggc tactcttggg agcgcaccat gaccttcgag gacaagggca
     6781 tcgtgaaggt gaagtccgac atctccatgg aggaggactc cttcatctac gagatacacc
     6841 tcaagggcga gaacttcccc cccaacggcc ccgtgatgca gaagaagacc accggctggg
     6901 acgcctccac cgagaggatg tacgtgcgcg acggcgtgct gaagggcgac gtcaagcaca
     6961 agctgctgct ggagggcggc ggccaccacc gcgttgactt caagaccatc tacagggcca
     7021 agaaggcggt gaagctgccc gactatcact ttgtggacca ccgcatcgag atcctgaacc
     7081 acgacaagga ctacaacaag gtgaccgttt acgagagcgc cgtggcccgc aactccaccg
     7141 acggcatgga cgagctgtac aaggcggccg cggagttccc agagcagaaa gcaggagaag
     7201 ctattaatca gcccatgatg atggctgcta ggcagttgca tgacgaggct cggaaatggt
     7261 ctagcaaggg taacgacatc attgctgctg ctaaacgaat ggcgctgctc atggctgaga
     7321 tgtcacgcct ggtgcgagga ggcagcggaa acaagcgtgc cctcatccag tgtgcaaaag
     7381 atattgctaa ggcatcggat gaagtcactc gattggccaa agaggtggcc aagcaatgta
     7441 ctgataaacg cattagaaca aacctcttac aggtctgtga gcgaatccca accatcagca
     7501 cgcagctcaa aattctttcc acagtgaaag ctaccatgct gggcaggact aacatcagcg
     7561 atgaagaatc agaacaggca actgagatgt tggttcataa cgcccagaac ctcatgcagg
     7621 atgtgaagga aactgtgaga gaagctgaag cagcatccat taagataaga acagatgccg
     7681 gattcactct gcgctgggtc agaaagaccc catggtatca ggaattcatg gtgagcaagg
     7741 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     7801 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     7861 tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     7921 tgggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     7981 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     8041 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     8101 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     8161 actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
     8221 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc
     8281 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     8341 agtccaaact gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     8401 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaatc gattctagag
     8461 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     8521 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     8581 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     8641 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     8701 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     8761 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     8821 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     8881 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     8941 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     9001 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     9061 ttaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     9121 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     9181 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     9241 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     9301 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     9361 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     9421 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     9481 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     9541 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     9601 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     9661 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     9721 gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg
     9781 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     9841 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     9901 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     9961 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
    10021 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
    10081 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
    10141 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
    10201 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
    10261 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
    10321 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
    10381 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
    10441 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
    10501 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
    10561 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
    10621 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
    10681 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
    10741 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
    10801 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
    10861 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
    10921 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
    10981 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
    11041 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
    11101 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
    11161 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
    11221 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
    11281 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
    11341 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
    11401 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
    11461 aaaggatcta ggtgaagatc ctttttga
//
