LOCUS       sequence-406245-        7094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GFP-tagged human integrin beta-3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7094)
  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 7094)
  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-3-GFP
FEATURES             Location/Qualifiers
     source          1..7094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3737..3746
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3743..3745,3746..3748,3749..4462)
                     /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(3743..3745,3746..3748,3749..4459)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3788..3809)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4049..4068)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4396..4417
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4585..4706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4622..4641)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4676..4695
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4713..5168)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4850..4871)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5062..5081
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5195..5299
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5301..5658
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5301..5492
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5324..5344)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5509..5644
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5693..6487
                     /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             5693..6487
                     /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(5747..5766)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6357..6376
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6675..6694)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6719..6766
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga
     1321 gctcaagctt gccaccatgc gagcgcggcc gcggccccgg ccgctctggg cgactgtgct
     1381 ggcgctgggg gcgctggcgg gcgttggcgt aggagggccc aacatctgta ccacgcgagg
     1441 tgtgagctcc tgccagcagt gcctggctgt gagccccatg tgtgcctggt gctctgatga
     1501 ggccctgcct ctgggctcac ctcgctgtga cctgaaggag aatctgctga aggataactg
     1561 tgccccagaa tccatcgagt tcccagtgag tgaggcccga gtactagagg acaggcccct
     1621 cagcgacaag ggctctggag acagctccca ggtcactcaa gtcagtcccc agaggattgc
     1681 actccggctc cggccagatg attcgaagaa tttctccatc caagtgcggc aggtggagga
     1741 ttaccctgtg gacatctact acttgatgga cctgtcttac tccatgaagg atgatctgtg
     1801 gagcatccag aacctgggta ccaagctggc cacccagatg cgaaagctca ccagtaacct
     1861 gcggattggc ttcggggcat ttgtggacaa gcctgtgtca ccatacatgt atatctcccc
     1921 accagaggcc ctcgaaaacc cctgctatga tatgaagacc acctgcttgc ccatgtttgg
     1981 ctacaaacac gtgctgacgc taactgacca ggtgacccgc ttcaatgagg aagtgaagaa
     2041 gcagagtgtg tcacggaacc gagatgcccc agagggtggc tttgatgcca tcatgcaggc
     2101 tacagtctgt gatgaaaaga ttggctggag gaatgatgca tcccacttgc tggtgtttac
     2161 cactgatgcc aagactcata tagcattgga cggaaggctg gcaggcattg tccagcctaa
     2221 tgacgggcag tgtcatgttg gtagtgacaa tcattactct gcctccacta ccatggatta
     2281 tccctctttg gggctgatga ctgagaagct atcccagaaa aacatcaatt tgatctttgc
     2341 agtgactgaa aatgtagtca atctctatca gaactatagt gagctcatcc cagggaccac
     2401 agttggggtt ctgtccatgg attccagcaa tgtcctccag ctcattgttg atgcttatgg
     2461 gaaaatccgt tctaaagtcg agctggaagt gcgtgacctc cctgaagagt tgtctctatc
     2521 cttcaatgcc acctgcctca acaatgaggt catccctggc ctcaagtctt gtatgggact
     2581 caagattgga gacacggtga gcttcagcat tgaggccaag gtgcgaggct gtccccagga
     2641 gaaggagaag tcctttacca taaagcccgt gggcttcaag gacagcctga tcgtccaggt
     2701 cacctttgat tgtgactgtg cctgccaggc ccaagctgaa cctaatagcc atcgctgcaa
     2761 caatggcaat gggacctttg agtgtggggt atgccgttgt gggcctggct ggctgggatc
     2821 ccagtgtgag tgctcagagg aggactatcg cccttcccag caggacgagt gcagcccccg
     2881 agagggtcag cccgtctgca gccagcgggg cgagtgcctc tgtggtcaat gtgtctgcca
     2941 cagcagtgac tttggcaaga tcacgggcaa gtactgcgag tgtgacgact tctcctgtgt
     3001 ccgctacaag ggggagatgt gctcaggcca tggccagtgc agctgtgggg actgcctgtg
     3061 tgactccgac tggaccggct actactgcaa ctgtaccacg cgtactgaca cctgcatgtc
     3121 cagcaatggg ctgctgtgca gcggccgcgg caagtgtgaa tgtggcagct gtgtctgtat
     3181 ccagccgggc tcctatgggg acacctgtga gaagtgcccc acctgcccag atgcctgcac
     3241 ctttaagaaa gaatgtgtgg agtgtaagaa gtttgaccgg ggagccctac atgacgaaaa
     3301 tacctgcaac cgttactgcc gtgacgagat tgagtcagtg aaagagctta aggacactgg
     3361 caaggatgca gtgaattgta cctataagaa tgaggatgac tgtgtcgtca gattccagta
     3421 ctatgaagat tctagtggaa agtccatcct gtatgtggta gaagagccag agtgtcccaa
     3481 gggccctgac atcctggtgg tcctgctctc agtgatgggg gccattctgc tcattggcct
     3541 tgccgccctg ctcatctgga aactcctcat caccatccac gaccgaaaag aattcgctaa
     3601 atttgaggaa gaacgcgcca gagcaaaatg ggacacagcc aacaacccac tgtataaaga
     3661 ggccacgtct accttcacca atatcacgta ccggggcact cagtcgacgg taccgcgggc
     3721 ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     3781 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3841 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3901 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     3961 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     4021 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     4081 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     4141 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     4201 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     4261 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     4321 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     4381 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     4441 catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca
     4501 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     4561 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     4621 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4681 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     4741 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     4801 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     4861 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4921 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4981 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5041 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5101 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     5161 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5221 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5281 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     5341 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5401 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5461 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5521 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5581 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5641 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     5701 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5761 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5821 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     5881 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5941 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     6001 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     6061 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     6121 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     6181 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     6241 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6301 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6361 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6421 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6481 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     6541 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     6601 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     6661 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     6721 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     6781 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     6841 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     6901 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     6961 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7021 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7081 atcaaaggat cttc
//
