LOCUS       sequence-403994-       10263 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expressing actin-binding-deficient vinculin mutant with 
            co-localization labeler TurboID and fluorescence reporter Venus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10263)
  AUTHORS   Tao A, LaCroix AS, Shoyer TC, Venkatraman V, Xu KL, Feiger B, 
            Hoffman BD
  TITLE     Identifying constitutive and context-specific 
            molecular-tension-sensitive protein recruitment within focal 
            adhesions.
  JOURNAL   Dev Cell. 2023 Mar 10:S1534-5807(23)00074-6. doi: 
            10.1016/j.devcel.2023.02.015.
  PUBMED    36924770
REFERENCE   2  (bases 1 to 10263)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dev Cell. 
            2023 Mar 10:S1534-5807(23)00074-6. doi: 
            10.1016/j.devcel.2023.02.015."
COMMENT     Sequence Label: pcDNA3.1-VinculinI997A_TurboID_Venus
FEATURES             Location/Qualifiers
     source          1..10263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4124..5083
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             join(5090..5092,5093..5095,5096..5809)
                     /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(5396..5415)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5743..5764
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5854..5871)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5860..6084
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6130..6558
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6217..6236)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6427..6448
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6567..6587)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6572..6901
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6752..6887
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6814..6833
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6968..7762
                     /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(7022..7041)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7632..7651
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7936..8057
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7973..7992)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8027..8046
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8106..8122)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8106..8122)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8119..8141)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8130..8146
                     /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(8154..8160,8161..8178,8179..8184))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8199..8220
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8337..8354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8508..9096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8588..8607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9267..10058,10059..10127))
                     /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"
     promoter        complement(10128..10232)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca tgcccgtctt ccacacgcgc accatcgaga gcatcttgga
      961 gcccgtggct cagcagatct cccacctggt catcatgcac gaggaggggg aggtagacgg
     1021 caaggccatc ccggacctca ccgcccccgt gtcggccgtg caggccgctg tcagcaacct
     1081 ggtgcgggtt ggaaaagaaa ctgtgcagac aacagaagac cagatcttga aaagggatat
     1141 gccaccagca ttcatcaaag tagagaatgc ctgcaccaag ctcgttcgag cagcccagat
     1201 gctgcaagca gatccttatt cagtaccagc tcgtgactac ctaattgatg gatcaagagg
     1261 catcctttct ggaacatcag acttacttct gacatttgat gaagcggagg tccgtaaaat
     1321 catccgtgtc tgcaaaggaa tattggaata tctgactgtg gcagaagtag tagagactat
     1381 ggaggatttg gtgacatata caaagaatct agggccagga atgacaaaga tggcgaaaat
     1441 gatcgatgag agacaacagg aattaactca tcaggaacat agggttatgc tggtgaactc
     1501 catgaatact gtgaaggagc tattgcctgt acttatttca gctatgaaga tctttgtaac
     1561 cacaaaaaac actaaaagcc agggaataga agaggccttg aaaaatcgca atttcacagt
     1621 agagaaaatg agtgctgaga taaatgaaat aatccgtgta ttacaactca cttcctggga
     1681 tgaagatgcc tgggccagca aggacactga agccatgaaa agagctttag ccctaataga
     1741 ttcaaagatg aaccaggcaa aaggctggct gagagatcca aacgcacctc caggggatgc
     1801 tggtgagcaa gcaataaggc agatccttga tgaagctgga aaagcaggag aattgtgtgc
     1861 aggcaaagaa cgcagggaga ttctgggaac atgcaaaact ttgggccaaa tgactgatca
     1921 acttgctgat ctccgagcta gaggacaggg tgctacaccg atggcaatgc agaaggcaca
     1981 gcaggtgtca caaggcctgg atttgctcac tgcaaaagtg gagaatgcag cccggaaatt
     2041 ggaggccatg acaaactcta agcaggctat tgcaaagaag attgatgctg ctcagaactg
     2101 gcttgcggat cccaacggcg gaagtgaagg agaagaacac attcgaggaa ttatgtctga
     2161 agcaaggaaa gttgcagaat tgtgtgagga gcctaaagaa agagatgata tccttcgctc
     2221 cttgggggaa atctctgctc tgacagctaa gctgtcagat ctgcgacgac atgggaaagg
     2281 cgactctcct gaggcccgtg cattggccaa gcaaatagct acatcacttc agaacttaca
     2341 gtccaaaaca aacagagctg tagcaaatac taggccagtt aaagctgctg tccatttgga
     2401 gggcaagatt gagcaagccc agaggtggat agataatcct acagttgatg atcggggagt
     2461 aggccaggca gcaattcggg gtttggttgc agaaggtcgt cgtctagcca atgtcatgat
     2521 gggaccttat cgtcaggacc tgcttgccaa atgtgaccgt gtagaccagc tggctgctca
     2581 gcttgctgac cttgcagcaa gaggggaagg agagtctcct caggctaggg caattgctgc
     2641 tcagcttcag gactccctga aggatctcaa agcacggatg caagaagcaa tgacccagga
     2701 ggtgtctgat gttttcagtg acactacaac ccctattaag ttgttagcag tagcagccac
     2761 tgctccttct gatactccca atagagaaga ggtgtttgaa gaaagagcag caaattttga
     2821 aaaccatgct gctagactgg gagcaacagc agaaaaagca gctgcagttg gaactgctaa
     2881 taaaactact gtggaaggca ttcaggcaac agtcaaatct gcaagggagc ttacaccaca
     2941 ggtagtatcg gctgctcgaa tcctcctgag aaatcctgga aatcaagctg cttatgagca
     3001 ttttgagaca atgaaaaacc aatggattga taatgtagaa aagatgacag ggctggtgga
     3061 tgaggccatc gataccaagt ctctgttgga tgcatcagaa gaggctatta agaaggatct
     3121 tgataaatgt aaagttgcaa tggccaatat gcaacctcag atgctggtag ctggagccac
     3181 cagcattgct agacgagcga accgcatcct gcttgtggca aaacgggagg ttgaaaattc
     3241 agaagaccct aaattcaggg aggctgttaa agcagcttct gatgagctga gcaaaaccat
     3301 atcaccgatg gtaatggatg ctaaagctgt agcaggaaat atctctgatc ctggtttgca
     3361 gaagagtttc ttggattctg gatacaggat tctgggagct gtggccaaag tcagagaagc
     3421 ctttcagcct caggagccag actttccccc tcctcctcct gacctagagc atctccatct
     3481 gactgatgag cttgctcctc caaaaccacc acttccagaa ggtgaggttc ccccacccag
     3541 accaccaccc cctgaagaaa aagatgagga gttcccagag cagaaagcag gagaagctat
     3601 taatcagccc atgatgatgg ctgctaggca gttgcatgac gaggctcgga aatggtctag
     3661 caagggtaac gacatcattg ctgctgctaa acgaatggcg ctgctcatgg ctgagatgtc
     3721 acgcctggtg cgaggaggca gcggaaacaa gcgtgccctc atccagtgtg caaaagatat
     3781 tgctaaggca tcggatgaag tcactcgatt ggccaaagag gtggccaagc aatgtactga
     3841 taaacgcatt agaacaaacc tcttacaggt ctgtgagcga atcccaacca tcagcacgca
     3901 gctcaaagct ctttccacag tgaaagctac catgctgggc aggactaaca tcagcgatga
     3961 agaatcagaa caggcaactg agatgttggt tcataacgcc cagaacctca tgcagtctgt
     4021 gaaggaaact gtgagagaag ctgaagcagc atccattaag ataagaacag atgccggatt
     4081 cactctgcgc tgggtcagaa agaccccatg gtatcaggaa ttcatgaaag acaatactgt
     4141 gcctctgaag ctgatcgctc tcctggctaa tggcgagttc catagtggcg aacagctggg
     4201 agaaaccctg ggcatgtcca gggccgctat caacaagcac attcagactc tgcgcgactg
     4261 gggcgtggac gtgttcaccg tgcccggaaa gggctactct ctgcccgagc ctatcccgct
     4321 gctgaacgct aaacagattc tgggacagct ggacggcggg agcgtggcag tcctgcctgt
     4381 ggtcgactcc accaatcagt acctgctgga tcgaatcggc gagctgaaga gtggggatgc
     4441 ttgcattgca gaatatcagc aggcagggag aggaagcaga gggaggaaat ggttctctcc
     4501 ttttggagct aacctgtacc tgagtatgtt ttggcgcctg aagcggggac cagcagcaat
     4561 cggcctgggc ccggtcatcg gaattgtcat ggcagaagcg ctgcgaaagc tgggagcaga
     4621 caaggtgcga gtcaaatggc ccaatgacct gtatctgcag gatagaaagc tggcaggcat
     4681 cctggtggag ctggccggaa taacaggcga tgctgcacag atcgtcattg gcgccgggat
     4741 taacgtggct atgaggcgcg tggaggaaag cgtggtcaat cagggctgga tcacactgca
     4801 ggaagcaggg attaacctgg acaggaatac tctggccgct acgctgatcc gagagctgcg
     4861 ggcagccctg gaactgttcg agcaggaagg cctggctcca tatctgccac ggtgggagaa
     4921 gctggataac ttcatcaata gacccgtgaa gctgatcatt ggggacaaag agattttcgg
     4981 gattagccgg gggattgata aacagggagc cctgctgctg gaacaggacg gagttatcaa
     5041 accctggatg ggcggagaaa tcagtctgcg gtctgccgaa aagggtacca tggtgagcaa
     5101 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     5161 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     5221 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     5281 cctgggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     5341 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     5401 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     5461 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     5521 caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     5581 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     5641 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     5701 ccagtccaaa ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     5761 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag cggccgctcg
     5821 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     5881 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     5941 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     6001 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     6061 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     6121 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     6181 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     6241 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     6301 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     6361 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     6421 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     6481 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     6541 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     6601 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     6661 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     6721 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     6781 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     6841 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     6901 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     6961 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     7021 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     7081 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     7141 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     7201 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     7261 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     7321 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     7381 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     7441 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     7501 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     7561 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     7621 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     7681 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     7741 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     7801 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     7861 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     7921 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     7981 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     8041 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     8101 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     8161 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     8221 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     8281 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     8341 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     8401 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     8461 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     8521 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     8581 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     8641 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     8701 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     8761 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     8821 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     8881 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     8941 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     9001 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     9061 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     9121 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     9181 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     9241 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     9301 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     9361 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     9421 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     9481 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     9541 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     9601 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     9661 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     9721 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     9781 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     9841 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     9901 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     9961 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
    10021 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
    10081 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
    10141 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
    10201 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
    10261 gtc
//
