LOCUS       sequence-361177-        6177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of talin1 head wt in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6177)
  AUTHORS   Azizi L, Varela L, Turkki P, Mykuliak VV, Korpela S, Ihalainen TO, 
            Church J, Hytonen VP, Goult BT
  TITLE     Talin variant P229S compromises integrin activation and associates 
            with multifaceted clinical symptoms.
  JOURNAL   Hum Mol Genet. 2022 Jul 21. pii: 6647920. doi: 10.1093/hmg/ddac163.
  PUBMED    35861643
REFERENCE   2  (bases 1 to 6177)
  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: "Hum Mol 
            Genet. 2022 Jul 21. pii: 6647920. doi: 10.1093/hmg/ddac163."
COMMENT     Sequence Label: talin1-head
FEATURES             Location/Qualifiers
     source          1..6177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      644..653
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2105..2107,2108..2110,2111..2824)
                     /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(2105..2107,2108..2110,2111..2821)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2150..2171)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2411..2430)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2758..2779
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2965..3086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3002..3021)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3056..3075
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3093..3548)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3230..3251)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3442..3461
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3575..3679
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3681..4038
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3681..3872
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3704..3724)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3889..4024
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4073..4867
                     /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             4073..4867
                     /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(4127..4146)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4737..4756
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5055..5074)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5099..5146
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5475..6063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5964..5983
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtattag gcaccccagg ctttacacga attcgcagaa gctgccacca tggttgcgct
      661 ttcgctgaag attagcattg ggaatgtggt gaagacgatg caatttgagc catctaccat
      721 ggtgtacgac gcctgccgca tgattcgtga gcggatccca gaggccctgg ctggccctcc
      781 caacgacttt gggctctttc tgtcagatga tgaccccaaa aaaggcatct ggctagaggc
      841 tgggaaagct ttggactact acatgctccg aaatggggac accatggagt acagaaaaaa
      901 gcagagaccc ctgaagatcc ggatgttaga cggaacggtg aagactatca tggtggatga
      961 ctccaagacc gtcacagaca tgctcatgac catttgtgcc cgaattggta tcaccaacca
     1021 tgatgagtat tcactggttc gagagctgat ggaagaaaag aaagatgagg ggacagggac
     1081 cctgagaaaa gacaagaccc tactgcgaga tgaaaagaag atggagaagc taaagcagaa
     1141 gttgcacaca gacgatgagt tgaactggct ggaccatggg cggacactga gggaacaggg
     1201 agtggaagag catgagacgt tgctgctgcg gagaaagttc ttctactcag accagaatgt
     1261 ggattcccga gaccctgtac agctgaacct tctctatgtg caggcacgag atgacatcct
     1321 gaatggctcc catcctgtct cctttgacaa ggcctgtgaa tttgcaggct tccagtgcca
     1381 gatccagttt ggacctcaca atgaacagaa gcacaaggct ggcttccttg acctgaagga
     1441 cttcctgccc aaggagtatg tgaagcagaa gggagagcgt aagatctttc aggcacacaa
     1501 gaattgtggg cagatgagtg aaattgaggc caaggtacgc tacgtgaagc tggcccgttc
     1561 cctcaagact tacggtgtct ccttcttcct agtcaaggaa aagatgaagg ggaagaataa
     1621 actggtgccc aggctgttgg gcatcactaa ggagtgtgtg atgcgtgtgg atgagaagac
     1681 caaggaggtg attcaggaat ggagcctcac caacatcaaa cgctgggctg cctctcccaa
     1741 gagctttact ctggactttg gagactatca ggatggctac tactcagtac agacgacaga
     1801 aggcgagcag atcgcacagc tcattgctgg ctacatagat atcatcctta agaagaaaaa
     1861 aagcaaggac cattttgggc tggagggaga tgaagagtct actatgttgg aggactcagt
     1921 gtctcccaaa aagtcaacag tccttcagca gcagtacaac cgagtgggga aagtggagca
     1981 cggctctgtg gctctgccag ccatcatgcg ctctggagcc tctggtcctg agaatttcca
     2041 ggtggggagc atgccacctg cccagcagca gatcaccagt ggccagatgc acggaggatc
     2101 cggaatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2161 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2221 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2281 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2341 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2401 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2461 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2521 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2581 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2641 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2701 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     2761 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2821 gtaaccgcgg gcccgggatc caccggatct agataactga tcataatcag ccataccaca
     2881 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     2941 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3001 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3061 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa
     3121 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3181 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3241 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3301 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3361 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3421 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3481 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3541 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3601 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3661 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3721 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3781 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3841 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3901 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3961 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4021 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4081 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4141 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4201 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4261 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4321 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4381 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4441 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4501 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4561 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4621 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4681 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4741 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4801 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4861 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4921 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4981 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     5041 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5101 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5161 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5221 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5281 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5341 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5401 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5461 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5521 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5581 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5641 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5701 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5761 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5821 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5881 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5941 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6001 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6061 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6121 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
