LOCUS       sequence-406246-        7127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GFP-tagged human integrin beta-5 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7127)
  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 7127)
  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: Integrin beta-5-GFP
FEATURES             Location/Qualifiers
     source          1..7127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      37..46
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(43..45,46..48,49..762)
                     /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(43..45,46..48,49..759)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(88..109)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(349..368)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     696..717
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    885..1006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(922..941)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     976..995
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1013..1468)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1150..1171)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1362..1381
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1495..1599
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1601..1958
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1601..1792
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1624..1644)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1809..1944
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1993..2787
                     /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             1993..2787
                     /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(2047..2066)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2657..2676
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2975..2994)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3019..3066
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3395..3983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3884..3903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4158..4461
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4462..4665
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4616..4636
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag
       61 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      121 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      181 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
      241 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
      301 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      361 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      421 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      481 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
      541 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
      601 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
      661 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
      721 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga
      781 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
      841 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
      901 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
      961 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     1021 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     1081 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     1141 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     1201 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     1261 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     1321 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     1381 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     1441 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     1501 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1561 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     1621 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     1681 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     1741 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     1801 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     1861 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     1921 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     1981 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     2041 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     2101 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     2161 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     2221 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     2281 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     2341 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     2401 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     2461 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     2521 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     2581 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     2641 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     2701 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     2761 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     2821 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     2881 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     2941 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     3001 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     3061 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     3121 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     3181 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     3241 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3301 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3361 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3421 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3481 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3541 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3601 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3661 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3721 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3781 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3841 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3901 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3961 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4021 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4081 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
     4141 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     4201 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     4261 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     4321 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     4381 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     4441 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     4501 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     4561 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     4621 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     4681 cagatccgct agcgctaccg gactcagatc tcgagctcaa gcttgccacc atgccgcggg
     4741 ccccggcgcc gctgtacgcc tgcctcctgg ggctctgcgc gctcctgccc cggctcgcag
     4801 gtctcaacat atgcactagt ggaagtgcca cctcatgtga agaatgtctg ctaatccacc
     4861 caaaatgtgc ctggtgctcc aaagaggact tcggaagccc acggtccatc acctctcggt
     4921 gtgatctgag ggcaaacctt gtcaaaaatg gctgtggagg tgagatagag agcccagcca
     4981 gcagcttcca tgtcctgagg agcctgcccc tcagcagcaa gggttcgggc tctgcaggct
     5041 gggacgtcat tcagatgaca ccacaggaga ttgccgtgaa cctccggccc ggtgacaaga
     5101 ccaccttcca gctacaggtt cgccaggtgg aggactatcc tgtggacctg tactacctga
     5161 tggacctctc cctgtccatg aaggatgact tggacaatat ccggagcctg ggcaccaaac
     5221 tcgcggagga gatgaggaag ctcaccagca acttccggtt gggatttggg tcttttgttg
     5281 ataaggacat ctctcctttc tcctacacgg caccgaggta ccagaccaat ccgtgcattg
     5341 gttacaagtt gtttccaaat tgcgtcccct cctttgggtt ccgccatctg ctgcctctca
     5401 cagacagagt ggacagcttc aatgaggaag ttcggaaaca gagggtgtcc cggaaccgag
     5461 atgcccctga ggggggcttt gatgcagtac tccaggcagc cgtctgcaag gagaagattg
     5521 gctggcgaaa ggatgcactg catttgctgg tgttcacaac agatgatgtg ccccacatcg
     5581 cattggatgg aaaattggga ggcctggtgc agccacacga tggccagtgc cacctgaacg
     5641 aggccaacga gtacactgca tccaaccaga tggactatcc atcccttgcc ttgcttggag
     5701 agaaattggc agagaacaac atcaacctca tctttgcagt gacaaaaaac cattatatgc
     5761 tgtacaagaa ttttacagcc ctgatacctg gaacaacggt ggagatttta gatggagact
     5821 ccaaaaatat tattcaactg attattaatg catacaatag tatccggtct aaagtggagt
     5881 tgtcagtctg ggatcagcct gaggatctta atctcttctt tactgctacc tgccaagatg
     5941 gggtatccta tcctggtcag aggaagtgtg agggtctgaa gattggggac acggcatctt
     6001 ttgaagtatc attggaggcc cgaagctgtc ccagcagaca cacggagcat gtgtttgccc
     6061 tgcggccggt gggattccgg gacagcctgg aggtgggggt cacctacaac tgcacgtgcg
     6121 gctgcagcgt ggggctggaa cccaacagcg ccaggtgcaa cgggagcggg acctatgtct
     6181 gcggcctgtg tgagtgcagc cccggctacc tgggcaccag gtgcgagtgc caggatgggg
     6241 agaaccagag cgtgtaccag aacctgtgcc gggaggcaga gggcaagcca ctgtgcagcg
     6301 ggcgtgggga ctgcagctgc aaccagtgct cctgcttcga gagcgagttt ggcaagatct
     6361 atgggccttt ctgtgagtgc gacaacttct cctgtgccag gaacaaggga gtcctctgct
     6421 caggccatgg cgagtgtcac tgcggggaat gcaagtgcca tgcaggttac atcggggaca
     6481 actgtaactg ctcgacagac atcagcacat gccggggcag agatggccag atctgcagcg
     6541 agcgtgggca ctgtctctgt gggcagtgcc aatgcacgga gccgggggcc tttggggaga
     6601 tgtgtgagaa gtgccccacc tgcccggatg catgcagcac caagagagat tgcgtcgagt
     6661 gcctgctgct ccactctggg aaacctgaca accagacctg ccacagccta tgcagggatg
     6721 aggtgatcac atgggtggac accatcgtga aagatgacca ggaggctgtg ctatgtttct
     6781 acaaaaccgc caaggactgc gtcatgatgt tcacctatgt ggagctcccc agtgggaagt
     6841 ccaacctgac cgtcctcagg gagccagagt gtggaaacac ccccaacgcc atgaccatcc
     6901 tcctggctgt ggtcggtagc atcctccttg ttgggcttgc actcctggct atctggaagc
     6961 tgcttgtcac catccacgac cggagggagt ttgcaaagtt tcagagcgag cgatccaggg
     7021 cccgctatga aatggcttca aatccattat acagaaagcc tatctccacg cacactgtgg
     7081 acttcacctt caacaagttc aacaaatcct acaatggcac tgtggac
//
