LOCUS       sequence-420344-       11635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vinculin tension sensor (VcnTS) with both directionally asymmetric, 
            force strengthening (DAFS) variant point mutations (E1015A and 
            E1021A), in lentiviral expression vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11635)
  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 11635)
  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: pRRL-VcnTS-E1015A-E1021A
FEATURES             Location/Qualifiers
     source          1..11635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      176..764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     665..684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     918..935
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1052..1073
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1088..1093,1094..1111,1112..1118)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1126..1142
                     /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     1131..1153
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1150..1166
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1150..1166
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1185..1205
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1187..1205
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1233..1459
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1460..1640
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1687..1812
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2305..2538
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2723..2767
                     /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    3034..3150
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3210..3589
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3590..3793
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3744..3764
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3838..3856
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6550..7257
                     /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(7378..7380,7381..7383,7384..8094)
                     /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(7423..7444)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7684..7703)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8031..8052
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    8707..9295
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8760..8780)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9178..9189)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9386..9619
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9691..9812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9728..9747)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9782..9801
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9852..9987
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(10008..10026)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10036..10053)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10036..10052)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10045..10067)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10194..10649
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10281..10300)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10491..10512
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10675..10779
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10780..10848,10849..11635,1..5)
                     /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(10998..11017)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
       61 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
      121 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
      181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      301 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
      361 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      421 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      481 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      541 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      601 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      661 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
      721 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
      781 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
      841 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
      901 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
      961 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     1021 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     1081 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     1141 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     1201 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
     1261 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
     1321 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
     1381 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
     1441 cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
     1501 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     1561 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     1621 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     1681 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     1741 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     1801 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     1861 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     1921 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     1981 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     2041 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     2101 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     2161 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     2221 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     2281 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     2341 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     2401 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     2461 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     2521 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     2581 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     2641 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     2701 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     2761 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     2821 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     2881 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     2941 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     3001 attagtgaac ggatctcgac ggtatcggtt aacttttaaa agaaaagggg ggattggggg
     3061 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     3121 acaaaaacaa attacaaaat tcaaaatttt atcgatggat cccccgggct cgagttcgaa
     3181 gatcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
     3241 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     3301 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     3361 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     3421 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     3481 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     3541 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     3601 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     3661 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     3721 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     3781 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
     3841 tacgactcac tatagggaga cccaagctgg ctagcgttta aacttaagct tgccatgccc
     3901 gtcttccaca cgcgcaccat cgagagcatc ttggagcccg tggctcagca gatctcccac
     3961 ctggtcatca tgcacgagga gggggaggta gacggcaagg ccatcccgga cctcaccgcc
     4021 cccgtgtcgg ccgtgcaggc cgctgtcagc aacctggtgc gggttggaaa agaaactgtg
     4081 cagacaacag aagaccagat cttgaaaagg gatatgccac cagcattcat caaagtagag
     4141 aatgcctgca ccaagctcgt tcgagcagcc cagatgctgc aagcagatcc ttattcagta
     4201 ccagctcgtg actacctaat tgatggatca agaggcatcc tttctggaac atcagactta
     4261 cttctgacat ttgatgaagc ggaggtccgt aaaatcatcc gtgtctgcaa aggaatattg
     4321 gaatatctga ctgtggcaga agtagtagag actatggagg atttggtgac atatacaaag
     4381 aatctagggc caggaatgac aaagatggcg aaaatgatcg atgagagaca acaggaatta
     4441 actcatcagg aacatagggt tatgctggtg aactccatga atactgtgaa ggagctattg
     4501 cctgtactta tttcagctat gaagatcttt gtaaccacaa aaaacactaa aagccaggga
     4561 atagaagagg ccttgaaaaa tcgcaatttc acagtagaga aaatgagtgc tgagataaat
     4621 gaaataatcc gtgtattaca actcacttcc tgggatgaag atgcctgggc cagcaaggac
     4681 actgaagcca tgaaaagagc tttagcccta atagattcaa agatgaacca ggcaaaaggc
     4741 tggctgagag atccaaacgc acctccaggg gatgctggtg agcaagcaat aaggcagatc
     4801 cttgatgaag ctggaaaagc aggagaattg tgtgcaggca aagaacgcag ggagattctg
     4861 ggaacatgca aaactttggg ccaaatgact gatcaacttg ctgatctccg agctagagga
     4921 cagggtgcta caccgatggc aatgcagaag gcacagcagg tgtcacaagg cctggatttg
     4981 ctcactgcaa aagtggagaa tgcagcccgg aaattggagg ccatgacaaa ctctaagcag
     5041 gctattgcaa agaagattga tgctgctcag aactggcttg cggatcccaa cggcggaagt
     5101 gaaggagaag aacacattcg aggaattatg tctgaagcaa ggaaagttgc agaattgtgt
     5161 gaggagccta aagaaagaga tgatatcctt cgctccttgg gggaaatctc tgctctgaca
     5221 gctaagctgt cagatctgcg acgacatggg aaaggcgact ctcctgaggc ccgtgcattg
     5281 gccaagcaaa tagctacatc acttcagaac ttacagtcca aaacaaacag agctgtagca
     5341 aatactaggc cagttaaagc tgctgtccat ttggagggca agattgagca agcccagagg
     5401 tggatagata atcctacagt tgatgatcgg ggagtaggcc aggcagcaat tcggggtttg
     5461 gttgcagaag gtcgtcgtct agccaatgtc atgatgggac cttatcgtca ggacctgctt
     5521 gccaaatgtg accgtgtaga ccagctggct gctcagcttg ctgaccttgc agcaagaggg
     5581 gaaggagagt ctcctcaggc tagggcaatt gctgctcagc ttcaggactc cctgaaggat
     5641 ctcaaagcac ggatgcaaga agcaatgacc caggaggtgt ctgatgtttt cagtgacact
     5701 acaaccccta ttaagttgtt agcagtagca gccactgctc cttctgatac tcccaataga
     5761 gaagaggtgt ttgaagaaag agcagcaaat tttgaaaacc atgctgctag actgggagca
     5821 acagcagaaa aagcagctgc agttggaact gctaataaaa ctactgtgga aggcattcag
     5881 gcaacagtca aatctgcaag ggagcttaca ccacaggtag tatcggctgc tcgaatcctc
     5941 ctgagaaatc ctggaaatca agctgcttat gagcattttg agacaatgaa aaaccaatgg
     6001 attgataatg tagaaaagat gacagggctg gtggatgagg ccatcgatac caagtctctg
     6061 ttggatgcat cagaagaggc tattaagaag gatcttgata aatgtaaagt tgcaatggcc
     6121 aatatgcaac ctcagatgct ggtagctgga gccaccagca ttgctagacg agcgaaccgc
     6181 atcctgcttg tggcaaaacg ggaggttgaa aattcagaag accctaaatt cagggaggct
     6241 gttaaagcag cttctgatga gctgagcaaa accatatcac cgatggtaat ggatgctaaa
     6301 gctgtagcag gaaatatctc tgatcctggt ttgcagaaga gtttcttgga ttctggatac
     6361 aggattctgg gagctgtggc caaagtcaga gaagcctttc agcctcagga gccagacttt
     6421 ccccctcctc ctcctgacct agagcatctc catctgactg atgagcttgc tcctccaaaa
     6481 ccaccacttc cagaaggtga ggttccccca cccagaccac caccccctga agaaaaagat
     6541 gaggtcgaga tggtgagcaa gggcgaggag accacaatgg gcgtaatcaa gcccgacatg
     6601 aagatcaagc tgaagatgga gggcaacgtg aatggccacg ccttcgtgat cgagggcgag
     6661 ggcgagggca agccctacga cggcaccaac accatcaacc tggaggtgaa ggagggagcc
     6721 cccctgccct tctcctacga cattctgacc accgcgttcg cctacggcaa cagggccttc
     6781 accaagtacc ccgacgacat ccccaactac ttcaagcagt ccttccccga gggctactct
     6841 tgggagcgca ccatgacctt cgaggacaag ggcatcgtga aggtgaagtc cgacatctcc
     6901 atggaggagg actccttcat ctacgagata cacctcaagg gcgagaactt cccccccaac
     6961 ggccccgtga tgcagaagaa gaccaccggc tgggacgcct ccaccgagag gatgtacgtg
     7021 cgcgacggcg tgctgaaggg cgacgtcaag cacaagctgc tgctggaggg cggcggccac
     7081 caccgcgttg acttcaagac catctacagg gccaagaagg cggtgaagct gcccgactat
     7141 cactttgtgg accaccgcat cgagatcctg aaccacgaca aggactacaa caaggtgacc
     7201 gtttacgaga gcgccgtggc ccgcaactcc accgacggca tggacgagct gtacaagggg
     7261 ccaggtggtg cagggccagg tggtgcaggg ccaggtggtg cagggccagg tggtgcaggg
     7321 cccggtggtg caggtccagg tggtgcaggt ccaggtggtg caggtccagg tggtgctatg
     7381 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     7441 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     7501 aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     7561 gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
     7621 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     7681 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     7741 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     7801 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
     7861 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
     7921 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     7981 ctgagctacc agtccaaact gagcaaagac cccaacgaga agcgcgatca catggtcctg
     8041 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caaggcggcc
     8101 gcggagttcc cagagcagaa agcaggagaa gctattaatc agcccatgat gatggctgct
     8161 aggcagttgc atgacgaggc tcggaaatgg tctagcaagg gtaacgacat cattgctgct
     8221 gctaaacgaa tggcgctgct catggctgag atgtcacgcc tggtgcgagg aggcagcgga
     8281 aacaagcgtg ccctcatcca gtgtgcaaaa gatattgcta aggcatcgga tgaagtcact
     8341 cgattggcca aagaggtggc caagcaatgt actgataaac gcattagaac aaacctctta
     8401 caggtctgtg agcgaatccc aaccatcagc acgcagctca aaattctttc cacagtgaaa
     8461 gctaccatgc tgggcaggac taacatcagc gatgaagcct cagaacaggc aactgctatg
     8521 ttggttcata acgcccagaa cctcatgcag tctgtgaagg aaactgtgag agaagctgaa
     8581 gcagcatcca ttaagataag aacagatgcc ggattcactc tgcgctgggt cagaaagacc
     8641 ccatggtatc agtaagaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga
     8701 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     8761 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     8821 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     8881 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     8941 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     9001 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     9061 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     9121 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     9181 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     9241 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     9301 attaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     9361 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     9421 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     9481 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     9541 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     9601 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     9661 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     9721 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     9781 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     9841 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     9901 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     9961 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
    10021 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
    10081 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
    10141 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
    10201 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
    10261 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
    10321 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
    10381 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
    10441 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
    10501 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
    10561 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
    10621 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
    10681 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
    10741 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
    10801 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
    10861 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
    10921 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
    10981 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
    11041 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
    11101 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
    11161 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
    11221 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
    11281 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
    11341 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
    11401 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
    11461 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
    11521 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
    11581 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcatt
//
