LOCUS       sequence-422214-       10160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vinculin tension sensor (VcnTS) with a single directionally 
            asymmetric, force strengthening (DAFS) variant point mutation 
            (E1015A)..
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-E1015A
FEATURES             Location/Qualifiers
     source          1..10160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(49..68)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        240..619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        620..823
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     774..794
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     868..887
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        868..886
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3580..4287
                     /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(4408..4410,4411..4413,4414..5124)
                     /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(4453..4474)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4714..4733)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5061..5082
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5756..5773)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5762..5986
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6032..6460
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6119..6138)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6329..6350
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6469..6489)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6474..6803
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6654..6789
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6716..6735
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6870..7664
                     /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(6924..6943)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7534..7553
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7838..7959
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7875..7894)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7929..7948
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8008..8024)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8008..8024)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8021..8043)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8032..8048
                     /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(8056..8062,8063..8080,8081..8086))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8101..8122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8239..8256)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8410..8998)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8490..8509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9169..9960,9961..10029))
                     /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     9792..9811
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10030..10134)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
       61 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
      121 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
      181 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
      241 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      301 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      361 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      421 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      481 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      541 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      601 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      661 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      721 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      781 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      841 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg
      901 ctagcgttta aacttaagct tgccatgccc gtcttccaca cgcgcaccat cgagagcatc
      961 ttggagcccg tggctcagca gatctcccac ctggtcatca tgcacgagga gggggaggta
     1021 gacggcaagg ccatcccgga cctcaccgcc cccgtgtcgg ccgtgcaggc cgctgtcagc
     1081 aacctggtgc gggttggaaa agaaactgtg cagacaacag aagaccagat cttgaaaagg
     1141 gatatgccac cagcattcat caaagtagag aatgcctgca ccaagctcgt tcgagcagcc
     1201 cagatgctgc aagcagatcc ttattcagta ccagctcgtg actacctaat tgatggatca
     1261 agaggcatcc tttctggaac atcagactta cttctgacat ttgatgaagc ggaggtccgt
     1321 aaaatcatcc gtgtctgcaa aggaatattg gaatatctga ctgtggcaga agtagtagag
     1381 actatggagg atttggtgac atatacaaag aatctagggc caggaatgac aaagatggcg
     1441 aaaatgatcg atgagagaca acaggaatta actcatcagg aacatagggt tatgctggtg
     1501 aactccatga atactgtgaa ggagctattg cctgtactta tttcagctat gaagatcttt
     1561 gtaaccacaa aaaacactaa aagccaggga atagaagagg ccttgaaaaa tcgcaatttc
     1621 acagtagaga aaatgagtgc tgagataaat gaaataatcc gtgtattaca actcacttcc
     1681 tgggatgaag atgcctgggc cagcaaggac actgaagcca tgaaaagagc tttagcccta
     1741 atagattcaa agatgaacca ggcaaaaggc tggctgagag atccaaacgc acctccaggg
     1801 gatgctggtg agcaagcaat aaggcagatc cttgatgaag ctggaaaagc aggagaattg
     1861 tgtgcaggca aagaacgcag ggagattctg ggaacatgca aaactttggg ccaaatgact
     1921 gatcaacttg ctgatctccg agctagagga cagggtgcta caccgatggc aatgcagaag
     1981 gcacagcagg tgtcacaagg cctggatttg ctcactgcaa aagtggagaa tgcagcccgg
     2041 aaattggagg ccatgacaaa ctctaagcag gctattgcaa agaagattga tgctgctcag
     2101 aactggcttg cggatcccaa cggcggaagt gaaggagaag aacacattcg aggaattatg
     2161 tctgaagcaa ggaaagttgc agaattgtgt gaggagccta aagaaagaga tgatatcctt
     2221 cgctccttgg gggaaatctc tgctctgaca gctaagctgt cagatctgcg acgacatggg
     2281 aaaggcgact ctcctgaggc ccgtgcattg gccaagcaaa tagctacatc acttcagaac
     2341 ttacagtcca aaacaaacag agctgtagca aatactaggc cagttaaagc tgctgtccat
     2401 ttggagggca agattgagca agcccagagg tggatagata atcctacagt tgatgatcgg
     2461 ggagtaggcc aggcagcaat tcggggtttg gttgcagaag gtcgtcgtct agccaatgtc
     2521 atgatgggac cttatcgtca ggacctgctt gccaaatgtg accgtgtaga ccagctggct
     2581 gctcagcttg ctgaccttgc agcaagaggg gaaggagagt ctcctcaggc tagggcaatt
     2641 gctgctcagc ttcaggactc cctgaaggat ctcaaagcac ggatgcaaga agcaatgacc
     2701 caggaggtgt ctgatgtttt cagtgacact acaaccccta ttaagttgtt agcagtagca
     2761 gccactgctc cttctgatac tcccaataga gaagaggtgt ttgaagaaag agcagcaaat
     2821 tttgaaaacc atgctgctag actgggagca acagcagaaa aagcagctgc agttggaact
     2881 gctaataaaa ctactgtgga aggcattcag gcaacagtca aatctgcaag ggagcttaca
     2941 ccacaggtag tatcggctgc tcgaatcctc ctgagaaatc ctggaaatca agctgcttat
     3001 gagcattttg agacaatgaa aaaccaatgg attgataatg tagaaaagat gacagggctg
     3061 gtggatgagg ccatcgatac caagtctctg ttggatgcat cagaagaggc tattaagaag
     3121 gatcttgata aatgtaaagt tgcaatggcc aatatgcaac ctcagatgct ggtagctgga
     3181 gccaccagca ttgctagacg agcgaaccgc atcctgcttg tggcaaaacg ggaggttgaa
     3241 aattcagaag accctaaatt cagggaggct gttaaagcag cttctgatga gctgagcaaa
     3301 accatatcac cgatggtaat ggatgctaaa gctgtagcag gaaatatctc tgatcctggt
     3361 ttgcagaaga gtttcttgga ttctggatac aggattctgg gagctgtggc caaagtcaga
     3421 gaagcctttc agcctcagga gccagacttt ccccctcctc ctcctgacct agagcatctc
     3481 catctgactg atgagcttgc tcctccaaaa ccaccacttc cagaaggtga ggttccccca
     3541 cccagaccac caccccctga agaaaaagat gaggtcgaga tggtgagcaa gggcgaggag
     3601 accacaatgg gcgtaatcaa gcccgacatg aagatcaagc tgaagatgga gggcaacgtg
     3661 aatggccacg ccttcgtgat cgagggcgag ggcgagggca agccctacga cggcaccaac
     3721 accatcaacc tggaggtgaa ggagggagcc cccctgccct tctcctacga cattctgacc
     3781 accgcgttcg cctacggcaa cagggccttc accaagtacc ccgacgacat ccccaactac
     3841 ttcaagcagt ccttccccga gggctactct tgggagcgca ccatgacctt cgaggacaag
     3901 ggcatcgtga aggtgaagtc cgacatctcc atggaggagg actccttcat ctacgagata
     3961 cacctcaagg gcgagaactt cccccccaac ggccccgtga tgcagaagaa gaccaccggc
     4021 tgggacgcct ccaccgagag gatgtacgtg cgcgacggcg tgctgaaggg cgacgtcaag
     4081 cacaagctgc tgctggaggg cggcggccac caccgcgttg acttcaagac catctacagg
     4141 gccaagaagg cggtgaagct gcccgactat cactttgtgg accaccgcat cgagatcctg
     4201 aaccacgaca aggactacaa caaggtgacc gtttacgaga gcgccgtggc ccgcaactcc
     4261 accgacggca tggacgagct gtacaagggg ccaggtggtg cagggccagg tggtgcaggg
     4321 ccaggtggtg cagggccagg tggtgcaggg cccggtggtg caggtccagg tggtgcaggt
     4381 ccaggtggtg caggtccagg tggtgctatg gtgagcaagg gcgaggagct gttcaccggg
     4441 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     4501 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     4561 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc
     4621 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     4681 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     4741 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     4801 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     4861 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     4921 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     4981 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaaact gagcaaagac
     5041 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     5101 ctcggcatgg acgagctgta caaggcggcc gcggagttcc cagagcagaa agcaggagaa
     5161 gctattaatc agcccatgat gatggctgct aggcagttgc atgacgaggc tcggaaatgg
     5221 tctagcaagg gtaacgacat cattgctgct gctaaacgaa tggcgctgct catggctgag
     5281 atgtcacgcc tggtgcgagg aggcagcgga aacaagcgtg ccctcatcca gtgtgcaaaa
     5341 gatattgcta aggcatcgga tgaagtcact cgattggcca aagaggtggc caagcaatgt
     5401 actgataaac gcattagaac aaacctctta caggtctgtg agcgaatccc aaccatcagc
     5461 acgcagctca aaattctttc cacagtgaaa gctaccatgc tgggcaggac taacatcagc
     5521 gatgaagcct cagaacaggc aactgagatg ttggttcata acgcccagaa cctcatgcag
     5581 tctgtgaagg aaactgtgag agaagctgaa gcagcatcca ttaagataag aacagatgcc
     5641 ggattcactc tgcgctgggt cagaaagacc ccatggtatc agtaagaatt ctgcagatat
     5701 ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     5761 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     5821 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     5881 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     5941 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     6001 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     6061 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6121 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6181 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6241 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6301 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     6361 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     6421 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     6481 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6541 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6601 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     6661 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6721 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     6781 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     6841 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     6901 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     6961 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     7021 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     7081 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     7141 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     7201 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     7261 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     7321 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     7381 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     7441 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     7501 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     7561 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     7621 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     7681 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     7741 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     7801 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     7861 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     7921 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     7981 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     8041 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     8101 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     8161 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     8221 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     8281 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     8341 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     8401 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     8461 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     8521 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     8581 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     8641 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     8701 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     8761 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     8821 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     8881 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8941 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     9001 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     9061 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     9121 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     9181 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     9241 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     9301 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     9361 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     9421 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     9481 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     9541 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     9601 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     9661 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     9721 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     9781 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9841 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9901 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9961 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
    10021 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
    10081 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
    10141 tccccgaaaa gtgccacctg
//
