LOCUS       sequence-402913-        8660 bp    DNA     circular SYN 15-APR-2025
DEFINITION  EGFP-Talin(DeltaR4-12).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8660)
  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 8660)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-Nterminal-EGFP
FEATURES             Location/Qualifiers
     source          1..8660
                     /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      604..613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(610..612,613..615,616..1326)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(610..612,613..615,616..1326)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(655..676)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(916..935)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1263..1284
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1375..1384
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5445..5566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5482..5501)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5536..5555
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5573..6028)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5710..5731)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5922..5941
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6055..6159
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6161..6518
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6161..6352
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6184..6204)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6369..6504
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6553..7347
                     /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             6553..7347
                     /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(6607..6626)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7217..7236
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7535..7554)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7579..7626
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7955..8543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8444..8463
                     /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 atcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      661 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
      721 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
      781 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
      841 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
      901 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
      961 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1021 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     1081 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     1141 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1201 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     1261 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1321 tacaagtccg gactcagatc tcgagctcaa gctagcttcg aattcgcaga agctgccacc
     1381 atggttgcgc tttcgctgaa gattagcatt gggaatgtgg tgaagacgat gcaatttgag
     1441 ccatctacca tggtgtacga cgcctgccgc atgattcgtg agcggatccc agaggccctg
     1501 gctggccctc ccaacgactt tgggctcttt ctgtcagatg atgaccccaa aaaaggcatc
     1561 tggctagagg ctgggaaagc tttggactac tacatgctcc gaaatgggga caccatggag
     1621 tacagaaaaa agcagagacc cctgaagata cggatgttag acggaacggt gaagactatc
     1681 atggtggatg actccaagac cgtcacagac atgctcatga ccatttgtgc ccgaattggt
     1741 atcaccaacc atgatgagta ttcactggtt cgagagctga tggaagaaaa gaaagatgag
     1801 gggacaggga ccctgagaaa agacaagacc ctactgcgag atgaaaagaa gatggagaag
     1861 ctaaagcaga agttgcacac agacgatgag ttgaactggc tggaccatgg gcggacactg
     1921 agggaacagg gagtggaaga gcatgagacg ttgctgctgc ggagaaagtt cttctactca
     1981 gaccagaatg tggattcccg agaccctgta cagctgaacc ttctctatgt gcaggcacga
     2041 gatgacatcc tgaatggctc ccatcctgtc tcctttgaca aggcctgtga atttgcaggc
     2101 ttccagtgcc agatccagtt tggacctcac aatgaacaga agcacaaggc tggcttcctt
     2161 gacctgaagg acttcctgcc caaggagtat gtgaagcaga agggagagcg taagatcttt
     2221 caggcacaca agaattgtgg gcagatgagt gaaattgagg ccaaggtacg ctacgtgaag
     2281 ctggcccgtt ccctcaagac ttacggtgtc tccttcttcc tagtcaagga aaagatgaag
     2341 gggaagaata aactggtgcc caggctgttg ggcatcacta aggagtgtgt gatgcgtgtg
     2401 gatgagaaga ccaaggaggt gattcaggaa tggagcctca ccaacatcaa acgctgggct
     2461 gcctctccca agagctttac tctggacttt ggagactatc aggatggcta ctactcagta
     2521 cagacgacag aaggcgagca gatcgcacag ctcattgctg gctacataga tatcatcctt
     2581 aagaagaaaa aaagcaagga ccattttggg ctggagggag atgaagagtc tactatgttg
     2641 gaggactcag tgtctcccaa aaagtcaaca gtccttcagc agcagtacaa ccgagtgggg
     2701 aaagtggagc acggctctgt ggctctgcca gccatcatgc gctctggagc ctctggtcct
     2761 gagaatttcc aggtggggag catgccacct gcccagcagc agatcaccag tggccagatg
     2821 caccgaggac acatgccacc tctgacttca gcccagcagg cgctcactgg aaccattaac
     2881 tccagcatgc aggctgtgca ggctgcccag gccactctgg atgactttga gactctaccc
     2941 cctcttggcc aggatgctgc ctccaaagct tggcgtaaga acaagatgga tgaatcaaag
     3001 catgagatcc actcccaggt agatgccatc acagctggca ctgcctctgt ggtaaacctg
     3061 acagcaggag atcctgcaga gacagactat acagcagtgg gctgtgcagt caccaccatc
     3121 tcttccaacc tgacggagat gtcccgtggg gtgaagctat tggctgccct gctggaggat
     3181 gaaggcggca atggccggcc ccttctgcaa gcagcaaagg gccttgcagg ggctgtgtcg
     3241 gaactgcttc gcagtgctca gcccgccagc gctgagcctc gtcagaacct gctgcaggca
     3301 gccgggaacg tgggccaggc cagtggggag ctgttgcagc aaattgggga aagtgacact
     3361 gacccccact tccaggatgt tctaatgcag ctagcaaacg cagtggcaag tgctgcagcc
     3421 gccctggtcc tcaaggccaa gagtgtggcc cagcgcacag aggattccgg gcttcagacc
     3481 caagtgattg ctgcagctac gcaatgtgct ctgtccactt cccagctggt ggcctgtacc
     3541 aaggtggtag cacctacaat cagctcacct gtgtgccaag aacagctagt cgaggctggg
     3601 cgactggtgg ccaaagctgt ggagggctgt gtgtctgcct cgcaagcagc tacagaagat
     3661 ggacagctgc ttcgaggggt aggagcagca gccacggctg tcacccaggc cctcaatgag
     3721 ctgctgcagc acgtgaaggc ccacgccacc ggagctgggc ctgccggtcg gtatgaccaa
     3781 gccactgata ccatcctcac cgtcactgaa aacatcttca gctccatggg tgatgctggg
     3841 gagatggtgc gccaggcccg catcctggcc caagccacct cagacctggt caatgctatc
     3901 aaggccgatg ctgaggggga gagtgatctg gagaactcta gaaagctcct gagtgctgcc
     3961 aagatcctcg ctgatgccac cgccaagatg gtggaggcgg ccaagggagc agccgcccac
     4021 cctgacagtg aggaacagca gcagcgactg cgtgaagcag ctgaggggct tcgcatggcc
     4081 accaatgcag ctgcgcagaa cgccatcaag aaggagtggg tggacccaga ggaccctact
     4141 gtcattgctg agaatgagct cttgggagcc gcagccgcca tcgaggctgc agccaagaag
     4201 ctggagcagc tgaagcccag ggccaaaccc aaggaggcgg atgagtcctt gaactttgag
     4261 gaacaaatcc tagaagctgc caagtccatc gctgcagcca ccagtgcact ggtaaaggct
     4321 gcgtcagcag cccagaggga gctggtggct caaggaaagg tgggcgccat tccagccaat
     4381 gcactggatg atgggcagtg gtcgcagggc ctcatttccg ctgcccgtat ggtggctgcg
     4441 gccaccaaca atctgtgtga agcagctaac gcagctgtcc agggccatgc tagccaggag
     4501 aaactcattt cctcagccaa gcaagtagct gcctccacag ctcagctcct ggtagcttgc
     4561 aaggtcaaag cagatcagga ctctgaggca atgaaacggc tccaggctgc tggcaatgca
     4621 gtgaagaggg cctcagataa cctggtaaag gcggcccaga aagctgcagc cttcgaagac
     4681 caggagaatg agacggtggt ggtgaaggag aaaatggttg ggggcattgc ccagattatc
     4741 gcagcacagg aagagatgct tcggaaggaa cgagagctgg aagaggctcg gaaaaagctc
     4801 gcccagatcc ggcagcagca gtacaagttc ttgccttcag agcttcgaga cgagcactaa
     4861 aaaaccggtg tatttaatgc agacccagcc cagagactgt gcctgccact accaaagcct
     4921 tctgggctgc tcaggaccca agctgcccaa ccccagcact cccccaaagt gcctgccaaa
     4981 ccctgggcct ggccctgccc agtcccactg caagccgtgt cctctccctg actcccaagt
     5041 gcctttgcac cctagggccc ctaagtgcct gcccccctcc agagtattaa cgctccaaga
     5101 gtattattaa tgctgccgta cctcagtttg aacctgccag ggccccagct gctccagcct
     5161 gccagcagct tccagccagt ccccacggcc acgtcagctc aactcatccc tttttgatac
     5221 tatatcccct acccagctac ctatggggct tgagggttgt aaacccaaac aggtcagact
     5281 cccgggattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     5341 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     5401 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     5461 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5521 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     5581 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5641 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5701 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5761 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     5821 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     5881 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     5941 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     6001 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     6061 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6121 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     6181 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     6241 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     6301 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     6361 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6421 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6481 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     6541 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     6601 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     6661 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     6721 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     6781 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     6841 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     6901 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     6961 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     7021 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     7081 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     7141 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     7201 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     7261 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     7321 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     7381 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     7441 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     7501 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     7561 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     7621 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     7681 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     7741 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     7801 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7861 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7921 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7981 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     8041 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     8101 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     8161 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     8221 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     8281 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8341 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8401 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8461 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8521 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8581 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8641 gtattaccgc catgcattag
//
