LOCUS       sequence-402937-        8395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Talin(DeltaR2-10)-L432G-mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8395)
  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 8395)
  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-T48-L432G-Cterminal-mEmerald
FEATURES             Location/Qualifiers
     source          1..8395
                     /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      4333..4342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4339..4341,4342..4344,4345..5058)
                     /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(4384..4405)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4645..4664)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4992..5013
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5180..5301
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5217..5236)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5271..5290
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5308..5763)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5445..5466)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5657..5676
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5790..5894
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5896..6253
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5896..6087
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5919..5939)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6104..6239
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6288..7082
                     /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             6288..7082
                     /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(6342..6361)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6952..6971
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7270..7289)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7314..7361
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7690..8278
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8179..8198
                     /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 aattggtacc caggcctgca ttacagcagc cagtgctgtg
     2641 tctggtatca ttgctgacct cgacaccacc atcatgttcg ctactgctgg cacacttaac
     2701 cgtgagggtg ctgaaacttt tgctgaccac cgggagggta tcttaaagac agcaaaggtt
     2761 cttgtggagg acaccaaggt cctagtgcag aatgcagctg ggagccagga gaagttggca
     2821 caagccgccc agtcctccgt ggccaccatt acccgcctcg ctgatgtggt caagctcggt
     2881 gcagccagcc taggagccga agaccctgaa actcaggtgg tgctgatcaa tgcagtaaag
     2941 gacgtagcca aggccctggg tgacctcatc agcgctacga aggctgcagc gggcaaagtt
     3001 ggggatgacc ctgcagtgtg gcagctcaag aactctgcca aggtgatggt gaccaatgtg
     3061 acatcattgc tcaagacagt gaaggctgtg gaagatgagg ccaccaaagg cacacgggcc
     3121 ctagaggcaa ccacagagca catacgtcag gaactggcgg tcttctgttc cccagagcca
     3181 cctgccaaga cctctacccc tgaagacttc atccgaatga ccaagggtat tactatggca
     3241 acagccaaag ccgttgctgc tggcaattcc tgtcgacagg aagatgtcat tgccacagcc
     3301 aatctgagcc gacgggctat tgcggacatg cttcgggctt gcaaggaagc agctttccac
     3361 ccagaagtgg cgcctgatgt acggctccga gccctgcact atggccggga gtgtgccaat
     3421 ggttacctgg aactgctgga ccacgtgctg ctgacccttc agaagccaaa cccagacctg
     3481 aagcagcagc taactggaca ctcaaagcgg gttgctggct ctgtgactga gctcatccag
     3541 gctgctgaag ccatgaaagg aacagagtgg gtggacccag aggaccctac tgtcattgct
     3601 gagaatgagc tcttgggagc cgcagccgcc atcgaggctg cagccaagaa gctggagcag
     3661 ctgaagccca gggccaaacc caaggaggcg gatgagtcct tgaactttga ggaacaaatc
     3721 ctagaagctg ccaagtccat cgctgcagcc accagtgcac tggtaaaggc tgcgtcagca
     3781 gcccagaggg agctggtggc tcaaggaaag gtgggcgcca ttccagccaa tgcactggat
     3841 gatgggcagt ggtcgcaggg cctcatttcc gctgcccgta tggtggctgc ggccaccaac
     3901 aatctgtgtg aagcagctaa cgcagctgtc cagggccatg ctagccagga gaaactcatt
     3961 tcctcagcca agcaagtagc tgcctccaca gctcagctcc tggtagcttg caaggtcaaa
     4021 gcagatcagg actctgaggc aatgaaacgg ctccaggctg ctggcaatgc agtgaagagg
     4081 gcctcagata acctggtaaa ggcggcccag aaagctgcag ccttcgaaga ccaggagaat
     4141 gagacggtgg tggtgaagga gaaaatggtt gggggcattg cccagattat cgcagcacag
     4201 gaagagatgc ttcggaagga acgagagctg gaagaggctc ggaaaaagct cgcccagatc
     4261 cggcagcagc agtacaagtt cttgccttca gagcttcgag acgagcacgg ctccgcgtcc
     4321 cgatcaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     4381 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     4441 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     4501 cccgtgccct ggcccaccct cgtgaccacc ttgacctacg gcgtgcagtg cttcgcccgc
     4561 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     4621 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     4681 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     4741 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaaggt ctatatcacc
     4801 gccgacaagc agaagaacgg catcaaggtg aacttcaaga cccgccacaa catcgaggac
     4861 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     4921 ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag
     4981 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     5041 gacgagctgt acaagtaagc ggccgcgact ctagatcata atcagccata ccacatttgt
     5101 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     5161 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     5221 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5281 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     5341 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     5401 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     5461 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     5521 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     5581 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     5641 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     5701 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     5761 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5821 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5881 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     5941 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     6001 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     6061 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     6121 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     6181 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     6241 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     6301 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     6361 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     6421 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     6481 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     6541 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     6601 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     6661 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     6721 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     6781 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     6841 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     6901 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     6961 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     7021 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     7081 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     7141 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     7201 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     7261 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     7321 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     7381 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     7441 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     7501 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     7561 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7621 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7681 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7741 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7801 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7861 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7921 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7981 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     8041 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     8101 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     8161 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     8221 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     8281 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     8341 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attag
//
