LOCUS       sequence-419258-       10772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vinculin (chicken) with S1033 phosphomimetic point mutation (S1033D)
            tagged with VenusA206K at the C-terminus, in lentiviral expression 
            vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10772)
  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 10772)
  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-VinculinVenus-S1033D
FEATURES             Location/Qualifiers
     source          1..10772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        21..400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        401..604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     555..575
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        649..667
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3910..3912,3913..3915,3916..4629)
                     /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(3955..3976)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4216..4235)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4563..4584
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4655..5243
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4708..4728)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5126..5137)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5334..5567
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5639..5760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5676..5695)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5730..5749
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5800..5935
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(5956..5974)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5984..6001)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5984..6000)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5993..6015)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6142..6597
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6229..6248)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6439..6460
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6623..6727
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6728..6796,6797..7588)
                     /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(6946..6965)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7759..8347
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8248..8267
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8501..8518
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8635..8656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8671..8676,8677..8694,8695..8701)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8709..8725
                     /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     8714..8736
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8733..8749
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8733..8749
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8768..8788
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8770..8788
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8816..9042
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9043..9223
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9270..9395
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9888..10121
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10306..10350
                     /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    10617..10733
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
       61 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      121 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      181 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      241 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      301 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      361 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      421 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      481 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      541 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      601 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
      661 ctatagggag acccaagctg gctagcgttt aaacttaagc ttgccatgcc cgtcttccac
      721 acgcgcacca tcgagagcat cttggagccc gtggctcagc agatctccca cctggtcatc
      781 atgcacgagg agggggaggt agacggcaag gccatcccgg acctcaccgc ccccgtgtcg
      841 gccgtgcagg ccgctgtcag caacctggtg cgggttggaa aagaaactgt gcagacaaca
      901 gaagaccaga tcttgaaaag ggatatgcca ccagcattca tcaaagtaga gaatgcctgc
      961 accaagctcg ttcgagcagc ccagatgctg caagcagatc cttattcagt accagctcgt
     1021 gactacctaa ttgatggatc aagaggcatc ctttctggaa catcagactt acttctgaca
     1081 tttgatgaag cggaggtccg taaaatcatc cgtgtctgca aaggaatatt ggaatatctg
     1141 actgtggcag aagtagtaga gactatggag gatttggtga catatacaaa gaatctaggg
     1201 ccaggaatga caaagatggc gaaaatgatc gatgagagac aacaggaatt aactcatcag
     1261 gaacataggg ttatgctggt gaactccatg aatactgtga aggagctatt gcctgtactt
     1321 atttcagcta tgaagatctt tgtaaccaca aaaaacacta aaagccaggg aatagaagag
     1381 gccttgaaaa atcgcaattt cacagtagag aaaatgagtg ctgagataaa tgaaataatc
     1441 cgtgtattac aactcacttc ctgggatgaa gatgcctggg ccagcaagga cactgaagcc
     1501 atgaaaagag ctttagccct aatagattca aagatgaacc aggcaaaagg ctggctgaga
     1561 gatccaaacg cacctccagg ggatgctggt gagcaagcaa taaggcagat ccttgatgaa
     1621 gctggaaaag caggagaatt gtgtgcaggc aaagaacgca gggagattct gggaacatgc
     1681 aaaactttgg gccaaatgac tgatcaactt gctgatctcc gagctagagg acagggtgct
     1741 acaccgatgg caatgcagaa ggcacagcag gtgtcacaag gcctggattt gctcactgca
     1801 aaagtggaga atgcagcccg gaaattggag gccatgacaa actctaagca ggctattgca
     1861 aagaagattg atgctgctca gaactggctt gcggatccca acggcggaag tgaaggagaa
     1921 gaacacattc gaggaattat gtctgaagca aggaaagttg cagaattgtg tgaggagcct
     1981 aaagaaagag atgatatcct tcgctccttg ggggaaatct ctgctctgac agctaagctg
     2041 tcagatctgc gacgacatgg gaaaggcgac tctcctgagg cccgtgcatt ggccaagcaa
     2101 atagctacat cacttcagaa cttacagtcc aaaacaaaca gagctgtagc aaatactagg
     2161 ccagttaaag ctgctgtcca tttggagggc aagattgagc aagcccagag gtggatagat
     2221 aatcctacag ttgatgatcg gggagtaggc caggcagcaa ttcggggttt ggttgcagaa
     2281 ggtcgtcgtc tagccaatgt catgatggga ccttatcgtc aggacctgct tgccaaatgt
     2341 gaccgtgtag accagctggc tgctcagctt gctgaccttg cagcaagagg ggaaggagag
     2401 tctcctcagg ctagggcaat tgctgctcag cttcaggact ccctgaagga tctcaaagca
     2461 cggatgcaag aagcaatgac ccaggaggtg tctgatgttt tcagtgacac tacaacccct
     2521 attaagttgt tagcagtagc agccactgct ccttctgata ctcccaatag agaagaggtg
     2581 tttgaagaaa gagcagcaaa ttttgaaaac catgctgcta gactgggagc aacagcagaa
     2641 aaagcagctg cagttggaac tgctaataaa actactgtgg aaggcattca ggcaacagtc
     2701 aaatctgcaa gggagcttac accacaggta gtatcggctg ctcgaatcct cctgagaaat
     2761 cctggaaatc aagctgctta tgagcatttt gagacaatga aaaaccaatg gattgataat
     2821 gtagaaaaga tgacagggct ggtggatgag gccatcgata ccaagtctct gttggatgca
     2881 tcagaagagg ctattaagaa ggatcttgat aaatgtaaag ttgcaatggc caatatgcaa
     2941 cctcagatgc tggtagctgg agccaccagc attgctagac gagcgaaccg catcctgctt
     3001 gtggcaaaac gggaggttga aaattcagaa gaccctaaat tcagggaggc tgttaaagca
     3061 gcttctgatg agctgagcaa aaccatatca ccgatggtaa tggatgctaa agctgtagca
     3121 ggaaatatct ctgatcctgg tttgcagaag agtttcttgg attctggata caggattctg
     3181 ggagctgtgg ccaaagtcag agaagccttt cagcctcagg agccagactt tccccctcct
     3241 cctcctgacc tagagcatct ccatctgact gatgagcttg ctcctccaaa accaccactt
     3301 ccagaaggtg aggttccccc acccagacca ccaccccctg aagaaaaaga tgaggagttc
     3361 ccagagcaga aagcaggaga agctattaat cagcccatga tgatggctgc taggcagttg
     3421 catgacgagg ctcggaaatg gtctagcaag ggtaacgaca tcattgctgc tgctaaacga
     3481 atggcgctgc tcatggctga gatgtcacgc ctggtgcgag gaggcagcgg aaacaagcgt
     3541 gccctcatcc agtgtgcaaa agatattgct aaggcatcgg atgaagtcac tcgattggcc
     3601 aaagaggtgg ccaagcaatg tactgataaa cgcattagaa caaacctctt acaggtctgt
     3661 gagcgaatcc caaccatcag cacgcagctc aaaattcttt ccacagtgaa agctaccatg
     3721 ctgggcagga ctaacatcag cgatgaagaa tcagaacagg caactgagat gttggttcat
     3781 aacgcccaga acctcatgca ggatgtgaag gaaactgtga gagaagctga agcagcatcc
     3841 attaagataa gaacagatgc cggattcact ctgcgctggg tcagaaagac cccatggtat
     3901 caggaattca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3961 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     4021 gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc
     4081 tggcccaccc tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg ctaccccgac
     4141 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     4201 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     4261 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     4321 ctggggcaca agctggagta caactacaac agccacaacg tctatatcac cgccgacaag
     4381 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     4441 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     4501 gacaaccact acctgagcta ccagtccaaa ctgagcaaag accccaacga gaagcgcgat
     4561 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     4621 tacaagtaag cggccgctcg agtctagagt cgacaatcaa cctctggatt acaaaatttg
     4681 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4741 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4801 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4861 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4921 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4981 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5041 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     5101 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5161 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     5221 ctccctttgg gccgcctccc cgcctggaat taattcgagc tcggtacctt taagaccaat
     5281 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     5341 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     5401 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     5461 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     5521 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     5581 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     5641 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5701 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5761 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
     5821 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5881 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg
     5941 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     6001 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     6061 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     6121 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6181 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     6241 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6301 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     6361 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6421 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6481 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     6541 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     6601 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6661 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6721 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6781 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6841 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6901 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6961 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     7021 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7081 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7141 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7201 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7261 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7321 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7381 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7441 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7501 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7561 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7621 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7681 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7741 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7801 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7861 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7921 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7981 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     8041 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     8101 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     8161 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     8221 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     8281 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     8341 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     8401 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     8461 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     8521 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     8581 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     8641 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     8701 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     8761 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag
     8821 tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta
     8881 caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc
     8941 ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat
     9001 tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac
     9061 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     9121 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     9181 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
     9241 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
     9301 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
     9361 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
     9421 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     9481 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     9541 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     9601 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     9661 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     9721 ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     9781 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     9841 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     9901 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     9961 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
    10021 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
    10081 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
    10141 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
    10201 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
    10261 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
    10321 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
    10381 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
    10441 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
    10501 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
    10561 agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg gttaactttt
    10621 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
    10681 acagacatac aaactaaaga attacaaaaa caaattacaa aattcaaaat tttatcgatg
    10741 gatcccccgg gctcgagttc gaagatcgat gt
//
