LOCUS       sequence-439051-        5945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Talin1 head EGFP fusion for mammalian 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5945)
  AUTHORS   Li J, Jo MH, Yan J, Hall T, Lee J, Lopez-Sanchez U, Yan S, Ha T, 
            Springer TA
  TITLE     Ligand binding initiates single-molecule integrin conformational 
            activation.
  JOURNAL   Cell. 2024 May 14:S0092-8674(24)00475-6. doi: 
            10.1016/j.cell.2024.04.049.
  PUBMED    38772370
REFERENCE   2  (bases 1 to 5945)
  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: "Cell. 2024 
            May 14:S0092-8674(24)00475-6. doi: 10.1016/j.cell.2024.04.049."
COMMENT     Sequence Label: Talin1 head EGFP fusion
FEATURES             Location/Qualifiers
     source          1..5945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..39)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      204..339
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             388..1182
                     /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             388..1182
                     /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(442..461)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1052..1071
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1370..1389)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1414..1461
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1790..2378
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2279..2298
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2553..2856
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2857..3060
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3011..3031
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3111..3120
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3115..3124
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4377..4386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4383..4385,4386..4388,4389..5102)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4383..4385,4386..4388,4389..5099)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4428..4449)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4689..4708)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5036..5057
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5225..5346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5262..5281)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5316..5335
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5353..5808)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5490..5511)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5702..5721
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5835..5939
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5941..5945,1..353)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        join(5941..5945,1..187)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
ORIGIN
        1 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
       61 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
      121 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      181 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
      241 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
      301 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
      361 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
      421 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
      481 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
      541 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
      601 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
      661 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
      721 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
      781 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
      841 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
      901 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
      961 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     1021 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     1081 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     1141 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     1201 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     1261 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     1321 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     1381 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     1441 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     1501 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     1561 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     1621 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1681 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1741 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1801 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1861 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1921 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1981 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2041 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2101 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2161 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2221 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2281 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2341 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2401 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2461 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
     2521 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     2581 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     2641 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     2701 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     2761 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     2821 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     2881 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     2941 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     3001 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     3061 ggtttagtga accgtcagat ccgctagcgc taccggactc agatctcgag gccaccatgg
     3121 tggcactttc actgaagatc agcattggga atgtggtgaa gacgatgcag tttgagccgt
     3181 ctaccatggt gtacgacgcc tgccgcatca ttcgtgagcg gatcccagag gccccagctg
     3241 gtcctcccag cgactttggg ctctttctgt cagatgatga ccccaaaaag ggtatatggc
     3301 tggaggctgg gaaagctttg gactactaca tgctccgaaa tggggacact atggagtaca
     3361 ggaagaaaca gagacccctg aagatccgta tgctggatgg aactgtgaag acgatcatgg
     3421 tggatgactc taagactgtc actgacatgc tcatgaccat ctgtgcccgc attggcatca
     3481 ccaatcatga tgaatattca ttggttcgag agctgatgga agagaaaaag gaggaaataa
     3541 cagggacctt aagaaaggac aagacattgc tgcgagatga aaagaagatg gagaaactaa
     3601 agcagaaatt gcacacagat gatgagttga actggctgga ccatggtcgg acactgaggg
     3661 agcagggtgt agaggagcac gagacgctgc tgctgcggag gaagttcttt tactcagacc
     3721 agaatgtgga ttcccgggac cctgtacagc tgaacctcct gtatgtgcag gcacgagatg
     3781 acatcctgaa tggctcccac cctgtctcct ttgacaaggc ctgtgagttt gctggcttcc
     3841 aatgccagat ccagtttggg ccccacaatg agcagaagca caaggctggc ttccttgacc
     3901 tgaaggactt cctgcccaag gagtatgtga agcagaaggg agagcgtaag atcttccagg
     3961 cacacaagaa ttgtgggcag atgagtgaga ttgaggccaa ggtccgctac gtgaagctag
     4021 cccgttctct caagacttac ggtgtctcct tcttcctggt gaaggaaaaa atgaaaggga
     4081 agaacaagct agtgcccagg cttctgggca tcaccaagga gtgtgtgatg cgagtggatg
     4141 agaagaccaa ggaagtgatc caggagtgga acctcaccaa catcaaacgc tgggctgcgt
     4201 ctcccaaaag cttcaccctg gattttggag attaccaaga tggctattac tcagtacaga
     4261 caactgaagg ggagcagatt gcacagctca ttgccggcta catcgatatc atcctgaaga
     4321 agaaaaaaag ccgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca
     4381 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     4441 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     4501 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     4561 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     4621 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     4681 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     4741 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     4801 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     4861 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     4921 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     4981 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     5041 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     5101 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     5161 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     5221 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5281 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5341 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     5401 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     5461 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     5521 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     5581 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     5641 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5701 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     5761 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     5821 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5881 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     5941 ctgag
//
