LOCUS       sequence-406236-        6445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GFP-tagged human zyxin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6445)
  AUTHORS   Zhang W, Lu CH, Nakamoto ML, Tsai CT, Roy AR, Lee CE, Yang Y, Jahed 
            Z, Li X, Cui B
  TITLE     Curved adhesions mediate cell attachment to soft matrix fibres in 
            three dimensions.
  JOURNAL   Nat Cell Biol. 2023 Sep 28. doi: 10.1038/s41556-023-01238-1.
  PUBMED    37770566
REFERENCE   2  (bases 1 to 6445)
  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: "Nat Cell 
            Biol. 2023 Sep 28. doi: 10.1038/s41556-023-01238-1."
COMMENT     Sequence Label: Zyxin-GFP
FEATURES             Location/Qualifiers
     source          1..6445
                     /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      631..640
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2385..2394
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2391..2393,2394..2396,2397..3110)
                     /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(2391..2393,2394..2396,2397..3107)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2436..2457)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2697..2716)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3044..3065
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3233..3354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3270..3289)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3324..3343
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3361..3816)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3498..3519)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3710..3729
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3843..3947
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3949..4306
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3949..4140
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3972..3992)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4157..4292
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4341..5135
                     /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             4341..5135
                     /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(4395..4414)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5005..5024
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5323..5342)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5367..5414
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5743..6331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6232..6251
                     /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 ccggactcag atctcgagct caagcttcgg ccatggcggc cccccgcccg tctcccgcga
      661 tctccgtttc ggtctcggct ccggcttttt acgccccgca gaagaagttc ggccctgtgg
      721 tggccccaaa gcccaaagtg aatcccttcc ggcccgggga cagcgagcct cccccggcac
      781 ccggggccca gcgcgcacag atgggccggg tgggcgagat tcccccgccg cccccggaag
      841 actttcccct gcctccacct ccccttgctg gggatggcga cgatgcagag ggtgctctgg
      901 gaggtgcctt cccgccgccc cctcccccga tcgaggaatc atttccccct gcgcctctgg
      961 aggaggagat cttcccttcc ccgccgcctc ctccggagga ggagggaggg cctgaggccc
     1021 ccataccgcc cccaccacag cccagggaga aggtgagcag tattgatttg gagatcgact
     1081 ctctgtcctc actgctggat gacatgacca agaatgatcc tttcaaagcc cgggtgtcat
     1141 ctggatatgt gcccccacca gtggccactc cattcagttc caagtccagt accaagcctg
     1201 cagccggggg cacagcaccc ctgcctcctt ggaagtcccc ttccagctcc cagcctctgc
     1261 cccaggttcc ggctccggct cagagccaga cacagttcca tgttcagccc cagccccagc
     1321 ccaagcctca ggtccaactc catgtccagt cccagaccca gcctgtgtct ttggctaaca
     1381 cccagccccg agggccccca gcctcatctc cggctccagc ccctaagttt tctccagtga
     1441 ctcctaagtt tactcctgtg gcttccaagt tcagtcctgg agccccaggt ggatctgggt
     1501 cacaaccaaa tcaaaaattg gggcaccccg aagctctttc tgctggcaca ggctcccctc
     1561 aacctcccag cttcacctat gcccagcaga gggagaagcc ccgagtgcag gagaagcagc
     1621 accccgtgcc cccaccggct cagaaccaaa accaggtgcg ctcccctggg gccccagggc
     1681 ccctgactct gaaggaggtg gaggagctgg agcagctgac ccagcagcta atgcaggaca
     1741 tggagcatcc tcagaggcag aatgtggctg tcaacgaact ctgcggccga tgccatcaac
     1801 ccctggcccg ggcgcagcca gccgtccgcg ctctagggca gctgttccac atcgcctgct
     1861 tcacctgcca ccagtgtgcg cagcagctcc agggccagca gttctacagt ctggaggggg
     1921 cgccgtactg cgagggctgt tacactgaca ccctggagaa gtgtaacacc tgcggggagc
     1981 ccatcactga ccgcatgctg agggccacgg gcaaggccta tcacccgcac tgcttcacct
     2041 gtgtggtctg cgcccgcccc ctggagggca cctccttcat cgtggaccag gccaaccggc
     2101 cccactgtgt ccccgactac cacaagcagt acgccccgag gtgctccgtc tgctctgagc
     2161 ccatcatgcc tgagcctggc cgagatgaga ctgtgcgagt ggtcgccctg gacaagaact
     2221 tccacatgaa gtgttacaag tgtgaggact gcgggaagcc cctgtcgatt gaggcagatg
     2281 acaatggctg cttccccctg gacggtcacg tgctctgtcg gaagtgccac actgctagag
     2341 cccagaccca gtcgacggta ccgcgggccc gggatccacc ggtcgccacc atggtgagca
     2401 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2461 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     2521 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2581 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2641 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2701 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2761 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2821 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     2881 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     2941 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     3001 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     3061 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg
     3121 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3181 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3241 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3301 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     3361 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3421 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3481 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3541 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3601 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3661 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3721 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3781 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3841 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3901 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3961 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4021 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4081 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4141 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4201 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4261 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4321 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4381 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4441 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4501 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4561 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4621 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4681 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4741 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4801 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4861 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4921 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4981 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5041 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5101 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5161 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5221 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5281 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5341 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5401 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5461 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5521 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5581 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5641 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5701 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5761 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5821 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5881 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5941 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6001 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6061 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6121 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6181 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6241 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6301 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6361 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6421 ggataaccgt attaccgcca tgcat
//
