LOCUS       sequence-422213-       10160 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Vinculin tension sensor (VcnTS) with both directionally asymmetric, 
            force strengthening (DAFS) variant point mutations (E1015A and 
            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 Apr 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-E1015A-E1021A
FEATURES             Location/Qualifiers
     source          1..10160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     866..885
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        866..884
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3578..4285
                     /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(4406..4408,4409..4411,4412..5122)
                     /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(4451..4472)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4712..4731)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5059..5080
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5754..5771)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5760..5984
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6030..6458
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6117..6136)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6327..6348
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6467..6487)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6472..6801
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6652..6787
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6714..6733
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6868..7662
                     /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(6922..6941)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7532..7551
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7836..7957
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7873..7892)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7927..7946
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8006..8022)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8006..8022)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8019..8041)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8030..8046
                     /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(8054..8060,8061..8078,8079..8084))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8099..8120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8237..8254)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8408..8996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8488..8507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9167..9958,9959..10027))
                     /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     9790..9809
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10028..10132)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
      841 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct
      901 agcgtttaaa cttaagcttg ccatgcccgt cttccacacg cgcaccatcg agagcatctt
      961 ggagcccgtg gctcagcaga tctcccacct ggtcatcatg cacgaggagg gggaggtaga
     1021 cggcaaggcc atcccggacc tcaccgcccc cgtgtcggcc gtgcaggccg ctgtcagcaa
     1081 cctggtgcgg gttggaaaag aaactgtgca gacaacagaa gaccagatct tgaaaaggga
     1141 tatgccacca gcattcatca aagtagagaa tgcctgcacc aagctcgttc gagcagccca
     1201 gatgctgcaa gcagatcctt attcagtacc agctcgtgac tacctaattg atggatcaag
     1261 aggcatcctt tctggaacat cagacttact tctgacattt gatgaagcgg aggtccgtaa
     1321 aatcatccgt gtctgcaaag gaatattgga atatctgact gtggcagaag tagtagagac
     1381 tatggaggat ttggtgacat atacaaagaa tctagggcca ggaatgacaa agatggcgaa
     1441 aatgatcgat gagagacaac aggaattaac tcatcaggaa catagggtta tgctggtgaa
     1501 ctccatgaat actgtgaagg agctattgcc tgtacttatt tcagctatga agatctttgt
     1561 aaccacaaaa aacactaaaa gccagggaat agaagaggcc ttgaaaaatc gcaatttcac
     1621 agtagagaaa atgagtgctg agataaatga aataatccgt gtattacaac tcacttcctg
     1681 ggatgaagat gcctgggcca gcaaggacac tgaagccatg aaaagagctt tagccctaat
     1741 agattcaaag atgaaccagg caaaaggctg gctgagagat ccaaacgcac ctccagggga
     1801 tgctggtgag caagcaataa ggcagatcct tgatgaagct ggaaaagcag gagaattgtg
     1861 tgcaggcaaa gaacgcaggg agattctggg aacatgcaaa actttgggcc aaatgactga
     1921 tcaacttgct gatctccgag ctagaggaca gggtgctaca ccgatggcaa tgcagaaggc
     1981 acagcaggtg tcacaaggcc tggatttgct cactgcaaaa gtggagaatg cagcccggaa
     2041 attggaggcc atgacaaact ctaagcaggc tattgcaaag aagattgatg ctgctcagaa
     2101 ctggcttgcg gatcccaacg gcggaagtga aggagaagaa cacattcgag gaattatgtc
     2161 tgaagcaagg aaagttgcag aattgtgtga ggagcctaaa gaaagagatg atatccttcg
     2221 ctccttgggg gaaatctctg ctctgacagc taagctgtca gatctgcgac gacatgggaa
     2281 aggcgactct cctgaggccc gtgcattggc caagcaaata gctacatcac ttcagaactt
     2341 acagtccaaa acaaacagag ctgtagcaaa tactaggcca gttaaagctg ctgtccattt
     2401 ggagggcaag attgagcaag cccagaggtg gatagataat cctacagttg atgatcgggg
     2461 agtaggccag gcagcaattc ggggtttggt tgcagaaggt cgtcgtctag ccaatgtcat
     2521 gatgggacct tatcgtcagg acctgcttgc caaatgtgac cgtgtagacc agctggctgc
     2581 tcagcttgct gaccttgcag caagagggga aggagagtct cctcaggcta gggcaattgc
     2641 tgctcagctt caggactccc tgaaggatct caaagcacgg atgcaagaag caatgaccca
     2701 ggaggtgtct gatgttttca gtgacactac aacccctatt aagttgttag cagtagcagc
     2761 cactgctcct tctgatactc ccaatagaga agaggtgttt gaagaaagag cagcaaattt
     2821 tgaaaaccat gctgctagac tgggagcaac agcagaaaaa gcagctgcag ttggaactgc
     2881 taataaaact actgtggaag gcattcaggc aacagtcaaa tctgcaaggg agcttacacc
     2941 acaggtagta tcggctgctc gaatcctcct gagaaatcct ggaaatcaag ctgcttatga
     3001 gcattttgag acaatgaaaa accaatggat tgataatgta gaaaagatga cagggctggt
     3061 ggatgaggcc atcgatacca agtctctgtt ggatgcatca gaagaggcta ttaagaagga
     3121 tcttgataaa tgtaaagttg caatggccaa tatgcaacct cagatgctgg tagctggagc
     3181 caccagcatt gctagacgag cgaaccgcat cctgcttgtg gcaaaacggg aggttgaaaa
     3241 ttcagaagac cctaaattca gggaggctgt taaagcagct tctgatgagc tgagcaaaac
     3301 catatcaccg atggtaatgg atgctaaagc tgtagcagga aatatctctg atcctggttt
     3361 gcagaagagt ttcttggatt ctggatacag gattctggga gctgtggcca aagtcagaga
     3421 agcctttcag cctcaggagc cagactttcc ccctcctcct cctgacctag agcatctcca
     3481 tctgactgat gagcttgctc ctccaaaacc accacttcca gaaggtgagg ttcccccacc
     3541 cagaccacca ccccctgaag aaaaagatga ggtcgagatg gtgagcaagg gcgaggagac
     3601 cacaatgggc gtaatcaagc ccgacatgaa gatcaagctg aagatggagg gcaacgtgaa
     3661 tggccacgcc ttcgtgatcg agggcgaggg cgagggcaag ccctacgacg gcaccaacac
     3721 catcaacctg gaggtgaagg agggagcccc cctgcccttc tcctacgaca ttctgaccac
     3781 cgcgttcgcc tacggcaaca gggccttcac caagtacccc gacgacatcc ccaactactt
     3841 caagcagtcc ttccccgagg gctactcttg ggagcgcacc atgaccttcg aggacaaggg
     3901 catcgtgaag gtgaagtccg acatctccat ggaggaggac tccttcatct acgagataca
     3961 cctcaagggc gagaacttcc cccccaacgg ccccgtgatg cagaagaaga ccaccggctg
     4021 ggacgcctcc accgagagga tgtacgtgcg cgacggcgtg ctgaagggcg acgtcaagca
     4081 caagctgctg ctggagggcg gcggccacca ccgcgttgac ttcaagacca tctacagggc
     4141 caagaaggcg gtgaagctgc ccgactatca ctttgtggac caccgcatcg agatcctgaa
     4201 ccacgacaag gactacaaca aggtgaccgt ttacgagagc gccgtggccc gcaactccac
     4261 cgacggcatg gacgagctgt acaaggggcc aggtggtgca gggccaggtg gtgcagggcc
     4321 aggtggtgca gggccaggtg gtgcagggcc cggtggtgca ggtccaggtg gtgcaggtcc
     4381 aggtggtgca ggtccaggtg gtgctatggt gagcaagggc gaggagctgt tcaccggggt
     4441 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     4501 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
     4561 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt
     4621 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     4681 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     4741 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     4801 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     4861 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     4921 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     4981 ccccgtgctg ctgcccgaca accactacct gagctaccag tccaaactga gcaaagaccc
     5041 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     5101 cggcatggac gagctgtaca aggcggccgc ggagttccca gagcagaaag caggagaagc
     5161 tattaatcag cccatgatga tggctgctag gcagttgcat gacgaggctc ggaaatggtc
     5221 tagcaagggt aacgacatca ttgctgctgc taaacgaatg gcgctgctca tggctgagat
     5281 gtcacgcctg gtgcgaggag gcagcggaaa caagcgtgcc ctcatccagt gtgcaaaaga
     5341 tattgctaag gcatcggatg aagtcactcg attggccaaa gaggtggcca agcaatgtac
     5401 tgataaacgc attagaacaa acctcttaca ggtctgtgag cgaatcccaa ccatcagcac
     5461 gcagctcaaa attctttcca cagtgaaagc taccatgctg ggcaggacta acatcagcga
     5521 tgaagcctca gaacaggcaa ctgctatgtt ggttcataac gcccagaacc tcatgcagtc
     5581 tgtgaaggaa actgtgagag aagctgaagc agcatccatt aagataagaa cagatgccgg
     5641 attcactctg cgctgggtca gaaagacccc atggtatcag taagaattct gcagatatcc
     5701 agcacagtgg cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     5761 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     5821 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     5881 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     5941 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     6001 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     6061 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     6121 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     6181 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     6241 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     6301 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     6361 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     6421 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     6481 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6541 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6601 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     6661 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6721 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     6781 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     6841 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     6901 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     6961 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     7021 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     7081 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     7141 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     7201 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     7261 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     7321 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     7381 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     7441 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     7501 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     7561 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     7621 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     7681 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     7741 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     7801 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     7861 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     7921 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     7981 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     8041 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     8101 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     8161 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     8221 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     8281 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     8341 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     8401 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     8461 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     8521 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     8581 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     8641 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     8701 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     8761 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     8821 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     8881 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     8941 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     9001 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     9061 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     9121 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     9181 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     9241 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     9301 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     9361 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     9421 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     9481 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     9541 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     9601 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     9661 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     9721 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     9781 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     9841 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     9901 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     9961 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
    10021 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
    10081 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
    10141 cccgaaaagt gccacctgac
//
