LOCUS       sequence-402897-        8203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Talin(DeltaR4-12)-L432G-mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8203)
  AUTHORS   Liu J, Wang Y, Goh WI, Goh H, Baird MA, Ruehland S, Teo S, Bate N, 
            Critchley DR, Davidson MW, Kanchanawong P
  TITLE     Talin determines the nanoscale architecture of focal adhesions.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Sep 1;112(35):E4864-73. doi: 
            10.1073/pnas.1512025112. Epub 2015 Aug 17.
  PUBMED    26283369
REFERENCE   2  (bases 1 to 8203)
  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"; doi: 
            "10.1073/pnas.1512025112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-09-1- 1"; volume: "112"; issue: "35"; pages: "E4864-73"
COMMENT     Sequence Label: Talin-T46-L432G-Cterminal-mEmerald
FEATURES             Location/Qualifiers
     source          1..8203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        58..361
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        362..565
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     516..536
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      634..643
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4141..4150
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4147..4149,4150..4152,4153..4866)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4192..4213)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4453..4472)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4800..4821
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4988..5109
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5025..5044)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5079..5098
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5116..5571)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5253..5274)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5465..5484
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5598..5702
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5704..6061
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5704..5895
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5727..5747)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5912..6047
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6096..6890
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6096..6890
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6150..6169)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6760..6779
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7078..7097)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7122..7169
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7498..8086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7987..8006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
       61 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      121 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      181 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      241 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      301 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      361 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      421 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      481 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      541 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
      601 gactcagatc tcgagctcaa gcttcgaatt ctagccacca tggttgcgct ttcgctgaag
      661 attagcattg ggaatgtggt gaagacgatg caatttgagc catctaccat ggtgtacgac
      721 gcctgccgca tgattcgtga gcggatccca gaggccctgg ctggccctcc caacgacttt
      781 gggctctttc tgtcagatga tgaccccaaa aaaggcatct ggctagaggc tgggaaagct
      841 ttggactact acatgctccg aaatggggac accatggagt acagaaaaaa gcagagaccc
      901 ctgaagatac ggatgttaga cggaacggtg aagactatca tggtggatga ctccaagacc
      961 gtcacagaca tgctcatgac catttgtgcc cgaattggta tcaccaacca tgatgagtat
     1021 tcactggttc gagagctgat ggaagaaaag aaagatgagg ggacagggac cctgagaaaa
     1081 gacaagaccc tactgcgaga tgaaaagaag atggagaagc taaagcagaa gttgcacaca
     1141 gacgatgagt tgaactggct ggaccatggg cggacactga gggaacaggg agtggaagag
     1201 catgagacgt tgctgctgcg gagaaagttc ttctactcag accagaatgt ggattcccga
     1261 gaccctgtac agctgaacct tctctatgtg caggcacgag atgacatcct gaatggctcc
     1321 catcctgtct cctttgacaa ggcctgtgaa tttgcaggct tccagtgcca gatccagttt
     1381 ggacctcaca atgaacagaa gcacaaggct ggcttccttg acctgaagga cttcctgccc
     1441 aaggagtatg tgaagcagaa gggagagcgt aagatctttc aggcacacaa gaattgtggg
     1501 cagatgagtg aaattgaggc caaggtacgc tacgtgaagc tggcccgttc cctcaagact
     1561 tacggtgtct ccttcttcct agtcaaggaa aagatgaagg ggaagaataa actggtgccc
     1621 aggctgttgg gcatcactaa ggagtgtgtg atgcgtgtgg atgagaagac caaggaggtg
     1681 attcaggaat ggagcctcac caacatcaaa cgctgggctg cctctcccaa gagctttact
     1741 ctggactttg gagactatca ggatggctac tactcagtac agacgacaga aggcgagcag
     1801 atcgcacagc tcattgctgg ctacatagat atcatcctta agaagaaaaa aagcaaggac
     1861 cattttgggc tggagggaga tgaagagtct actatgttgg aggactcagt gtctcccaaa
     1921 aagtcaacag tcggtcagca gcagtacaac cgagtgggga aagtggagca cggctctgtg
     1981 gctctgccag ccatcatgcg ctctggagcc tctggtcctg agaatttcca ggtggggagc
     2041 atgccacctg cccagcagca gatcaccagt ggccagatgc accgaggaca catgccacct
     2101 ctgacttcag cccagcaggc gctcactgga accattaact ccagcatgca ggctgtgcag
     2161 gctgcccagg ccactctgga tgactttgag actctacccc ctcttggcca ggatgctgcc
     2221 tccaaagctt ggcgtaagaa caagatggat gaatcaaagc atgagatcca ctcccaggta
     2281 gatgccatca cagctggcac tgcctctgtg gtaaacctga cagcaggaga tcctgcagag
     2341 acagactata cagcagtggg ctgtgcagtc accaccatct cttccaacct gacggagatg
     2401 tcccgtgggg tgaagctatt ggctgccctg ctggaggatg aaggcggcaa tggccggccc
     2461 cttctgcaag cagcaaaggg ccttgcaggg gctgtgtcgg aactgcttcg cagtgctcag
     2521 cccgccagcg ctgagcctcg tcagaacctg ctgcaggcag ccgggaacgt gggccaggcc
     2581 agtggggagc tgttgcagca aattggggaa agtgacactg acccccactt ccaggatgtt
     2641 ctaatgcagc tagcaaacgc agtggcaagt gctgcagccg ccctggtcct caaggccaag
     2701 agtgtggccc agcgcacaga ggattccggg cttcagaccc aagtgattgc tgcagctacg
     2761 caatgtgctc tgtccacttc ccagctggtg gcctgtacca aggtggtagc acctacaatc
     2821 agctcacctg tgtgccaaga acagctagtc gaggctgggc gactggtggc caaagctgtg
     2881 gagggctgtg tgtctgcctc gcaagcagct acagaagatg gacagctgct tcgaggggta
     2941 ggagcagcag ccacggctgt cacccaggcc ctcaatgagc tgctgcagca cgtgaaggcc
     3001 cacgccaccg gagctgggcc tgccggtcgg tatgaccaag ccactgatac catcctcacc
     3061 gtcactgaaa acatcttcag ctccatgggt gatgctgggg agatggtgcg ccaggcccgc
     3121 atcctggccc aagccacctc agacctggtc aatgctatca aggccgatgc tgagggggag
     3181 agtgatctgg agaactctag aaagctcctg agtgctgcca agatcctcgc tgatgccacc
     3241 gccaagatgg tggaggcggc caagggagca gccgcccacc ctgacagtga ggaacagcag
     3301 cagcgactgc gtgaagcagc tgaggggctt cgcatggcca ccaatgcagc tgcgcagaac
     3361 gccatcaaga aggagtgggt ggacccagag gaccctactg tcattgctga gaatgagctc
     3421 ttgggagccg cagccgccat cgaggctgca gccaagaagc tggagcagct gaagcccagg
     3481 gccaaaccca aggaggcgga tgagtccttg aactttgagg aacaaatcct agaagctgcc
     3541 aagtccatcg ctgcagccac cagtgcactg gtaaaggctg cgtcagcagc ccagagggag
     3601 ctggtggctc aaggaaaggt gggcgccatt ccagccaatg cactggatga tgggcagtgg
     3661 tcgcagggcc tcatttccgc tgcccgtatg gtggctgcgg ccaccaacaa tctgtgtgaa
     3721 gcagctaacg cagctgtcca gggccatgct agccaggaga aactcatttc ctcagccaag
     3781 caagtagctg cctccacagc tcagctcctg gtagcttgca aggtcaaagc agatcaggac
     3841 tctgaggcaa tgaaacggct ccaggctgct ggcaatgcag tgaagagggc ctcagataac
     3901 ctggtaaagg cggcccagaa agctgcagcc ttcgaagacc aggagaatga gacggtggtg
     3961 gtgaaggaga aaatggttgg gggcattgcc cagattatcg cagcacagga agagatgctt
     4021 cggaaggaac gagagctgga agaggctcgg aaaaagctcg cccagatccg gcagcagcag
     4081 tacaagttct tgccttcaga gcttcgagac gagcacggct ccgcgtcccg atcaccggtc
     4141 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     4201 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     4261 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     4321 cccaccctcg tgaccacctt gacctacggc gtgcagtgct tcgcccgcta ccccgaccac
     4381 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     4441 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     4501 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     4561 gggcacaagc tggagtacaa ctacaacagc cacaaggtct atatcaccgc cgacaagcag
     4621 aagaacggca tcaaggtgaa cttcaagacc cgccacaaca tcgaggacgg cagcgtgcag
     4681 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     4741 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     4801 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     4861 aagtaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     4921 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4981 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5041 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5101 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     5161 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     5221 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     5281 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     5341 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     5401 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     5461 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     5521 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     5581 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5641 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5701 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     5761 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     5821 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5881 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5941 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     6001 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     6061 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     6121 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     6181 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     6241 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     6301 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     6361 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     6421 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     6481 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     6541 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     6601 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     6661 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     6721 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     6781 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6841 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6901 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6961 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     7021 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     7081 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     7141 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     7201 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     7261 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     7321 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     7381 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7441 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7501 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7561 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7621 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7681 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7741 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7801 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7861 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7921 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7981 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     8041 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     8101 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     8161 atcccctgat tctgtggata accgtattac cgccatgcat tag
//
