LOCUS       sequence-457354-       10160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10160)
  AUTHORS   Grashoff C, Hoffman BD, Brenner MD, Zhou R, Parsons M, Yang MT, 
            McLean MA, Sligar SG, Chen CS, Ha T, Schwartz MA
  TITLE     Measuring mechanical tension across vinculin reveals regulation of 
            focal adhesion dynamics.
  JOURNAL   Nature. 2010 Jul 8. 466(7303):263-6.
  PUBMED    20613844
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: "Nature. 
            2010 Jul 8. 466(7303):263-6."
COMMENT     Sequence Label: VinculinTS
FEATURES             Location/Qualifiers
     source          1..10160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..29)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      823..1411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1312..1331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1565..1582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1699..1720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1735..1740,1741..1758,1759..1765)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1773..1789
                     /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     1778..1800
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1797..1813
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1797..1813
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1862..1983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1873..1892)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1927..1946
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2157..2951)
                     /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(2268..2287)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2878..2897
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3018..3347)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3032..3167
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3086..3105)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3332..3352
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3361..3789)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3471..3492)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3683..3702
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3835..4059
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4048..4065
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(4697..5407,5408..5410,5411..5413))
                     /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(4739..4760)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5088..5107
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5347..5368
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(5534..6241)
                     /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"
     primer_bind     complement(8934..8953)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8935..8953)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(8998..9201)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(9027..9047)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        9202..9581
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     9753..9772
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        9847..9951
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9952..10020,10021..10160,1..652)
                     /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"
ORIGIN
        1 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
       61 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
      121 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
      181 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
      241 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
      301 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
      361 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
      421 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
      481 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
      541 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
      601 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
      661 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
      721 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
      781 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
      841 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
      901 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
      961 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1021 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     1081 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     1141 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     1201 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     1261 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     1321 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     1381 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     1441 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     1501 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     1561 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     1621 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     1681 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     1741 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     1801 aaacagctat gaccatgatt acgccaagct ctagctagag gtcgacggta tacagacatg
     1861 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     1921 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     1981 gttggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc
     2041 gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc
     2101 tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag
     2161 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     2221 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     2281 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     2341 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
     2401 agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc
     2461 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     2521 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     2581 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     2641 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     2701 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     2761 gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     2821 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     2881 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     2941 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagatccgaa aatggatata
     3001 caagctcccg ggagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     3061 tggaatagct cagaggcaga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     3121 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     3181 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     3241 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     3301 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagaatt
     3361 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3421 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3481 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3541 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3601 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3661 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3721 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3781 agggcgcgtg gggatacccc ctagagcccc agctggttct ttccgcctca gaagccatag
     3841 agcccaccgc atccccagca tgcctgctat tgtcttccca atcctccccc ttgctgtcct
     3901 gccccacccc accccccaga atagaatgac acctactcag acaatgcgat gcaatttcct
     3961 cattttatta ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa ggcacggggg
     4021 aggggcaaac aacagatggc tggcaactag aaggcacagt cgaggctgat cagcgggttt
     4081 aaacgggccc tctagactcg agcggccgcc actgtgctgg atatctgcag aattcttact
     4141 gataccatgg ggtctttctg acccagcgca gagtgaatcc ggcatctgtt cttatcttaa
     4201 tggatgctgc ttcagcttct ctcacagttt ccttcacaga ctgcatgagg ttctgggcgt
     4261 tatgaaccaa catctcagtt gcctgttctg attcttcatc gctgatgtta gtcctgccca
     4321 gcatggtagc tttcactgtg gaaagaattt tgagctgcgt gctgatggtt gggattcgct
     4381 cacagacctg taagaggttt gttctaatgc gtttatcagt acattgcttg gccacctctt
     4441 tggccaatcg agtgacttca tccgatgcct tagcaatatc ttttgcacac tggatgaggg
     4501 cacgcttgtt tccgctgcct cctcgcacca ggcgtgacat ctcagccatg agcagcgcca
     4561 ttcgtttagc agcagcaatg atgtcgttac ccttgctaga ccatttccga gcctcgtcat
     4621 gcaactgcct agcagccatc atcatgggct gattaatagc ttctcctgct ttctgctctg
     4681 ggaactccgc ggccgccttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     4741 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agtttggact
     4801 ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     4861 gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt
     4921 tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg ctgttgtagt
     4981 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     5041 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     5101 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     5161 agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcagg ccgtagccca
     5221 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atcagcttca
     5281 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     5341 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     5401 ccttgctcac catagcacca cctggacctg caccacctgg acctgcacca cctggacctg
     5461 caccaccggg ccctgcacca cctggccctg caccacctgg ccctgcacca cctggccctg
     5521 caccacctgg ccccttgtac agctcgtcca tgccgtcggt ggagttgcgg gccacggcgc
     5581 tctcgtaaac ggtcaccttg ttgtagtcct tgtcgtggtt caggatctcg atgcggtggt
     5641 ccacaaagtg atagtcgggc agcttcaccg ccttcttggc cctgtagatg gtcttgaagt
     5701 caacgcggtg gtggccgccg ccctccagca gcagcttgtg cttgacgtcg cccttcagca
     5761 cgccgtcgcg cacgtacatc ctctcggtgg aggcgtccca gccggtggtc ttcttctgca
     5821 tcacggggcc gttggggggg aagttctcgc ccttgaggtg tatctcgtag atgaaggagt
     5881 cctcctccat ggagatgtcg gacttcacct tcacgatgcc cttgtcctcg aaggtcatgg
     5941 tgcgctccca agagtagccc tcggggaagg actgcttgaa gtagttgggg atgtcgtcgg
     6001 ggtacttggt gaaggccctg ttgccgtagg cgaacgcggt ggtcagaatg tcgtaggaga
     6061 agggcagggg ggctccctcc ttcacctcca ggttgatggt gttggtgccg tcgtagggct
     6121 tgccctcgcc ctcgccctcg atcacgaagg cgtggccatt cacgttgccc tccatcttca
     6181 gcttgatctt catgtcgggc ttgattacgc ccattgtggt ctcctcgccc ttgctcacca
     6241 tctcgacctc atctttttct tcagggggtg gtggtctggg tgggggaacc tcaccttctg
     6301 gaagtggtgg ttttggagga gcaagctcat cagtcagatg gagatgctct aggtcaggag
     6361 gaggaggggg aaagtctggc tcctgaggct gaaaggcttc tctgactttg gccacagctc
     6421 ccagaatcct gtatccagaa tccaagaaac tcttctgcaa accaggatca gagatatttc
     6481 ctgctacagc tttagcatcc attaccatcg gtgatatggt tttgctcagc tcatcagaag
     6541 ctgctttaac agcctccctg aatttagggt cttctgaatt ttcaacctcc cgttttgcca
     6601 caagcaggat gcggttcgct cgtctagcaa tgctggtggc tccagctacc agcatctgag
     6661 gttgcatatt ggccattgca actttacatt tatcaagatc cttcttaata gcctcttctg
     6721 atgcatccaa cagagacttg gtatcgatgg cctcatccac cagccctgtc atcttttcta
     6781 cattatcaat ccattggttt ttcattgtct caaaatgctc ataagcagct tgatttccag
     6841 gatttctcag gaggattcga gcagccgata ctacctgtgg tgtaagctcc cttgcagatt
     6901 tgactgttgc ctgaatgcct tccacagtag ttttattagc agttccaact gcagctgctt
     6961 tttctgctgt tgctcccagt ctagcagcat ggttttcaaa atttgctgct ctttcttcaa
     7021 acacctcttc tctattggga gtatcagaag gagcagtggc tgctactgct aacaacttaa
     7081 taggggttgt agtgtcactg aaaacatcag acacctcctg ggtcattgct tcttgcatcc
     7141 gtgctttgag atccttcagg gagtcctgaa gctgagcagc aattgcccta gcctgaggag
     7201 actctccttc ccctcttgct gcaaggtcag caagctgagc agccagctgg tctacacggt
     7261 cacatttggc aagcaggtcc tgacgataag gtcccatcat gacattggct agacgacgac
     7321 cttctgcaac caaaccccga attgctgcct ggcctactcc ccgatcatca actgtaggat
     7381 tatctatcca cctctgggct tgctcaatct tgccctccaa atggacagca gctttaactg
     7441 gcctagtatt tgctacagct ctgtttgttt tggactgtaa gttctgaagt gatgtagcta
     7501 tttgcttggc caatgcacgg gcctcaggag agtcgccttt cccatgtcgt cgcagatctg
     7561 acagcttagc tgtcagagca gagatttccc ccaaggagcg aaggatatca tctctttctt
     7621 taggctcctc acacaattct gcaactttcc ttgcttcaga cataattcct cgaatgtgtt
     7681 cttctccttc acttccgccg ttgggatccg caagccagtt ctgagcagca tcaatcttct
     7741 ttgcaatagc ctgcttagag tttgtcatgg cctccaattt ccgggctgca ttctccactt
     7801 ttgcagtgag caaatccagg ccttgtgaca cctgctgtgc cttctgcatt gccatcggtg
     7861 tagcaccctg tcctctagct cggagatcag caagttgatc agtcatttgg cccaaagttt
     7921 tgcatgttcc cagaatctcc ctgcgttctt tgcctgcaca caattctcct gcttttccag
     7981 cttcatcaag gatctgcctt attgcttgct caccagcatc ccctggaggt gcgtttggat
     8041 ctctcagcca gccttttgcc tggttcatct ttgaatctat tagggctaaa gctcttttca
     8101 tggcttcagt gtccttgctg gcccaggcat cttcatccca ggaagtgagt tgtaatacac
     8161 ggattatttc atttatctca gcactcattt tctctactgt gaaattgcga tttttcaagg
     8221 cctcttctat tccctggctt ttagtgtttt ttgtggttac aaagatcttc atagctgaaa
     8281 taagtacagg caatagctcc ttcacagtat tcatggagtt caccagcata accctatgtt
     8341 cctgatgagt taattcctgt tgtctctcat cgatcatttt cgccatcttt gtcattcctg
     8401 gccctagatt ctttgtatat gtcaccaaat cctccatagt ctctactact tctgccacag
     8461 tcagatattc caatattcct ttgcagacac ggatgatttt acggacctcc gcttcatcaa
     8521 atgtcagaag taagtctgat gttccagaaa ggatgcctct tgatccatca attaggtagt
     8581 cacgagctgg tactgaataa ggatctgctt gcagcatctg ggctgctcga acgagcttgg
     8641 tgcaggcatt ctctactttg atgaatgctg gtggcatatc ccttttcaag atctggtctt
     8701 ctgttgtctg cacagtttct tttccaaccc gcaccaggtt gctgacagcg gcctgcacgg
     8761 ccgacacggg ggcggtgagg tccgggatgg ccttgccgtc tacctccccc tcctcgtgca
     8821 tgatgaccag gtgggagatc tgctgagcca cgggctccaa gatgctctcg atggtgcgcg
     8881 tgtggaagac gggcatggca agcttaagtt taaacgctag ccagcttggg tctccctata
     8941 gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta gccagagagc
     9001 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     9061 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     9121 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     9181 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     9241 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     9301 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     9361 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     9421 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     9481 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     9541 cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat atctggcccg
     9601 tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc aattgtcggt
     9661 caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga cctccaacac
     9721 acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     9781 gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac ttttcgggga
     9841 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     9901 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     9961 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
    10021 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
    10081 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
    10141 tttccaatga tgagcacttt
//
