LOCUS       sequence-422049-       10160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vinculin tension sensor (VcnTS) with a single directionally 
            asymmetric, force strengthening (DAFS) variant point mutation 
            (E1021A)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10160)
  AUTHORS   Chirasani VR, Khan MAI, Malavade JN, Dokholyan NV, Hoffman BD, 
            Campbell SL
  TITLE     Molecular basis and cellular functions of vinculin-actin directional
            catch bonding.
  JOURNAL   Nat Commun. 2023 Dec 14;14(1):8300. doi: 10.1038/s41467-023-43779-x.
  PUBMED    38097542
REFERENCE   2  (bases 1 to 10160)
  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: "2023-12-14"; volume: "14(1):8300. doi"; pages: " 
            10.1038/s41467-023-43779-x"
COMMENT     Sequence Label: pcDNA3.1-VcnTS-E1021A
FEATURES             Location/Qualifiers
     source          1..10160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3577..4284
                     /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(4405..4407,4408..4410,4411..5121)
                     /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(4450..4471)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4711..4730)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5058..5079
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5753..5770)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5759..5983
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6029..6457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6116..6135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6326..6347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6466..6486)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6471..6800
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6651..6786
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6713..6732
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6867..7661
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6921..6940)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7531..7550
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7835..7956
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7872..7891)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7926..7945
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8005..8021)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8005..8021)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8018..8040)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8029..8045
                     /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(8053..8059,8060..8077,8078..8083))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8098..8119
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8236..8253)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8407..8995)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8487..8506)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9166..9957,9958..10026))
                     /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     9789..9808
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10027..10131)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gcgtttaaac ttaagcttgc catgcccgtc ttccacacgc gcaccatcga gagcatcttg
      961 gagcccgtgg ctcagcagat ctcccacctg gtcatcatgc acgaggaggg ggaggtagac
     1021 ggcaaggcca tcccggacct caccgccccc gtgtcggccg tgcaggccgc tgtcagcaac
     1081 ctggtgcggg ttggaaaaga aactgtgcag acaacagaag accagatctt gaaaagggat
     1141 atgccaccag cattcatcaa agtagagaat gcctgcacca agctcgttcg agcagcccag
     1201 atgctgcaag cagatcctta ttcagtacca gctcgtgact acctaattga tggatcaaga
     1261 ggcatccttt ctggaacatc agacttactt ctgacatttg atgaagcgga ggtccgtaaa
     1321 atcatccgtg tctgcaaagg aatattggaa tatctgactg tggcagaagt agtagagact
     1381 atggaggatt tggtgacata tacaaagaat ctagggccag gaatgacaaa gatggcgaaa
     1441 atgatcgatg agagacaaca ggaattaact catcaggaac atagggttat gctggtgaac
     1501 tccatgaata ctgtgaagga gctattgcct gtacttattt cagctatgaa gatctttgta
     1561 accacaaaaa acactaaaag ccagggaata gaagaggcct tgaaaaatcg caatttcaca
     1621 gtagagaaaa tgagtgctga gataaatgaa ataatccgtg tattacaact cacttcctgg
     1681 gatgaagatg cctgggccag caaggacact gaagccatga aaagagcttt agccctaata
     1741 gattcaaaga tgaaccaggc aaaaggctgg ctgagagatc caaacgcacc tccaggggat
     1801 gctggtgagc aagcaataag gcagatcctt gatgaagctg gaaaagcagg agaattgtgt
     1861 gcaggcaaag aacgcaggga gattctggga acatgcaaaa ctttgggcca aatgactgat
     1921 caacttgctg atctccgagc tagaggacag ggtgctacac cgatggcaat gcagaaggca
     1981 cagcaggtgt cacaaggcct ggatttgctc actgcaaaag tggagaatgc agcccggaaa
     2041 ttggaggcca tgacaaactc taagcaggct attgcaaaga agattgatgc tgctcagaac
     2101 tggcttgcgg atcccaacgg cggaagtgaa ggagaagaac acattcgagg aattatgtct
     2161 gaagcaagga aagttgcaga attgtgtgag gagcctaaag aaagagatga tatccttcgc
     2221 tccttggggg aaatctctgc tctgacagct aagctgtcag atctgcgacg acatgggaaa
     2281 ggcgactctc ctgaggcccg tgcattggcc aagcaaatag ctacatcact tcagaactta
     2341 cagtccaaaa caaacagagc tgtagcaaat actaggccag ttaaagctgc tgtccatttg
     2401 gagggcaaga ttgagcaagc ccagaggtgg atagataatc ctacagttga tgatcgggga
     2461 gtaggccagg cagcaattcg gggtttggtt gcagaaggtc gtcgtctagc caatgtcatg
     2521 atgggacctt atcgtcagga cctgcttgcc aaatgtgacc gtgtagacca gctggctgct
     2581 cagcttgctg accttgcagc aagaggggaa ggagagtctc ctcaggctag ggcaattgct
     2641 gctcagcttc aggactccct gaaggatctc aaagcacgga tgcaagaagc aatgacccag
     2701 gaggtgtctg atgttttcag tgacactaca acccctatta agttgttagc agtagcagcc
     2761 actgctcctt ctgatactcc caatagagaa gaggtgtttg aagaaagagc agcaaatttt
     2821 gaaaaccatg ctgctagact gggagcaaca gcagaaaaag cagctgcagt tggaactgct
     2881 aataaaacta ctgtggaagg cattcaggca acagtcaaat ctgcaaggga gcttacacca
     2941 caggtagtat cggctgctcg aatcctcctg agaaatcctg gaaatcaagc tgcttatgag
     3001 cattttgaga caatgaaaaa ccaatggatt gataatgtag aaaagatgac agggctggtg
     3061 gatgaggcca tcgataccaa gtctctgttg gatgcatcag aagaggctat taagaaggat
     3121 cttgataaat gtaaagttgc aatggccaat atgcaacctc agatgctggt agctggagcc
     3181 accagcattg ctagacgagc gaaccgcatc ctgcttgtgg caaaacggga ggttgaaaat
     3241 tcagaagacc ctaaattcag ggaggctgtt aaagcagctt ctgatgagct gagcaaaacc
     3301 atatcaccga tggtaatgga tgctaaagct gtagcaggaa atatctctga tcctggtttg
     3361 cagaagagtt tcttggattc tggatacagg attctgggag ctgtggccaa agtcagagaa
     3421 gcctttcagc ctcaggagcc agactttccc cctcctcctc ctgacctaga gcatctccat
     3481 ctgactgatg agcttgctcc tccaaaacca ccacttccag aaggtgaggt tcccccaccc
     3541 agaccaccac cccctgaaga aaaagatgag gtcgagatgg tgagcaaggg cgaggagacc
     3601 acaatgggcg taatcaagcc cgacatgaag atcaagctga agatggaggg caacgtgaat
     3661 ggccacgcct tcgtgatcga gggcgagggc gagggcaagc cctacgacgg caccaacacc
     3721 atcaacctgg aggtgaagga gggagccccc ctgcccttct cctacgacat tctgaccacc
     3781 gcgttcgcct acggcaacag ggccttcacc aagtaccccg acgacatccc caactacttc
     3841 aagcagtcct tccccgaggg ctactcttgg gagcgcacca tgaccttcga ggacaagggc
     3901 atcgtgaagg tgaagtccga catctccatg gaggaggact ccttcatcta cgagatacac
     3961 ctcaagggcg agaacttccc ccccaacggc cccgtgatgc agaagaagac caccggctgg
     4021 gacgcctcca ccgagaggat gtacgtgcgc gacggcgtgc tgaagggcga cgtcaagcac
     4081 aagctgctgc tggagggcgg cggccaccac cgcgttgact tcaagaccat ctacagggcc
     4141 aagaaggcgg tgaagctgcc cgactatcac tttgtggacc accgcatcga gatcctgaac
     4201 cacgacaagg actacaacaa ggtgaccgtt tacgagagcg ccgtggcccg caactccacc
     4261 gacggcatgg acgagctgta caaggggcca ggtggtgcag ggccaggtgg tgcagggcca
     4321 ggtggtgcag ggccaggtgg tgcagggccc ggtggtgcag gtccaggtgg tgcaggtcca
     4381 ggtggtgcag gtccaggtgg tgctatggtg agcaagggcg aggagctgtt caccggggtg
     4441 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     4501 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
     4561 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc
     4621 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     4681 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     4741 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     4801 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     4861 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     4921 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     4981 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaaactgag caaagacccc
     5041 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     5101 ggcatggacg agctgtacaa ggcggccgcg gagttcccag agcagaaagc aggagaagct
     5161 attaatcagc ccatgatgat ggctgctagg cagttgcatg acgaggctcg gaaatggtct
     5221 agcaagggta acgacatcat tgctgctgct aaacgaatgg cgctgctcat ggctgagatg
     5281 tcacgcctgg tgcgaggagg cagcggaaac aagcgtgccc tcatccagtg tgcaaaagat
     5341 attgctaagg catcggatga agtcactcga ttggccaaag aggtggccaa gcaatgtact
     5401 gataaacgca ttagaacaaa cctcttacag gtctgtgagc gaatcccaac catcagcacg
     5461 cagctcaaaa ttctttccac agtgaaagct accatgctgg gcaggactaa catcagcgat
     5521 gaagaatcag aacaggcaac tgctatgttg gttcataacg cccagaacct catgcagtct
     5581 gtgaaggaaa ctgtgagaga agctgaagca gcatccatta agataagaac agatgccgga
     5641 ttcactctgc gctgggtcag aaagacccca tggtatcagt aagaattctg cagatatcca
     5701 gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     5761 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     5821 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     5881 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     5941 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     6001 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     6061 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     6121 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     6181 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     6241 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     6301 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     6361 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     6421 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     6481 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     6541 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     6601 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     6661 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     6721 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     6781 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     6841 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     6901 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     6961 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     7021 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     7081 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     7141 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     7201 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     7261 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     7321 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     7381 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     7441 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     7501 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     7561 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     7621 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     7681 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     7741 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     7801 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     7861 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     7921 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     7981 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     8041 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     8101 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     8161 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     8221 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     8281 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     8341 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     8401 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     8461 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     8521 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     8581 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     8641 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     8701 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     8761 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     8821 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     8881 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     8941 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     9001 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     9061 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     9121 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     9181 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     9241 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     9301 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     9361 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     9421 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     9481 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     9541 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     9601 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     9661 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     9721 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     9781 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     9841 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     9901 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     9961 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
    10021 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
    10081 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
    10141 ccgaaaagtg ccacctgacg
//
