LOCUS       sequence-292325-        7027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Detecting Integrin beta5 by fluorescence microscopy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7027)
  AUTHORS   Lock JG, Jones MC, Askari JA, Gong X, Oddone A, Olofsson H, 
            Goransson S, Lakadamyali M, Humphries MJ, Stromblad S
  TITLE     Reticular adhesions are a distinct class of cell-matrix adhesions 
            that mediate attachment during mitosis.
  JOURNAL   Nat Cell Biol. 2018 Nov;20(11):1290-1302. doi: 
            10.1038/s41556-018-0220-2. Epub 2018 Oct 22.
  PUBMED    30361699
REFERENCE   2  (bases 1 to 7027)
  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"; doi: 
            "10.1038/s41556-018-0220-2"; journalName: "Nat Cell Biol"; date: 
            "2018-11"; volume: "20"; issue: "11"; pages: "1290-1302"
COMMENT     Sequence Label: Integrin beta5-2XEGFP
FEATURES             Location/Qualifiers
     source          1..7027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(2914..2916,2917..2919,2920..3633)
                     /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(2914..2916,2917..2919,2920..3630)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2959..2980)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3220..3239)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3567..3588
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3756..3877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3793..3812)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3847..3866
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3884..4339)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4021..4042)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4233..4252
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4366..4470
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4472..4829
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4472..4663
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4495..4515)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4680..4815
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4864..5658
                     /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             4864..5658
                     /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(4918..4937)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5528..5547
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5846..5865)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5890..5937
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6266..6854
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6755..6774
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcatg
      541 ccgcgggccc cggcgccgct gtacgcctgc ctcctggggc tctgcgcgct cctgccccgg
      601 ctcgcaggtc tcaacatatg cactagtgga agtgccacct catgtgaaga atgtctgcta
      661 atccacccaa aatgtgcctg gtgctccaaa gaggacttcg gaagcccacg gtccatcacc
      721 tctcggtgtg atctgagggc aaaccttgtc aaaaatggct gtggaggtga gatagagagc
      781 ccagccagca gcttccatgt cctgaggagc ctgcccctca gcagcaaggg ttcgggctct
      841 gcaggctggg acgtcattca gatgacacca caggagattg ccgtgaacct ccggcccggt
      901 gacaagacca ccttccagct acaggttcgc caggtggagg actatcctgt ggacctgtac
      961 tacctgatgg acctctccct gtccatgaag gatgacttgg acaatatccg gagcctgggc
     1021 accaaactcg cggaggagat gaggaagctc accagcaact tccggttggg atttgggtct
     1081 tttgttgata aggacatctc tcctttctcc tacacggcac cgaggtacca gaccaatccg
     1141 tgcattggtt acaagttgtt tccaaattgc gtcccctcct ttgggttccg ccatctgctg
     1201 cctctcacag acagagtgga cagcttcaat gaggaagttc ggaaacagag ggtgtcccgg
     1261 aaccgagatg cccctgaggg gggctttgat gcagtactcc aggcagccgt ctgcaaggag
     1321 aagattggct ggcgaaagga tgcactgcat ttgctggtgt tcacaacaga tgatgtgccc
     1381 cacatcgcat tggatggaaa attgggaggc ctggtgcagc cacacgatgg ccagtgccac
     1441 ctgaacgagg ccaacgagta cactgcatcc aaccagatgg actatccatc ccttgccttg
     1501 cttggagaga aattggcaga gaacaacatc aacctcatct ttgcagtgac aaaaaaccat
     1561 tatatgctgt acaagaattt tacagccctg atacctggaa caacggtgga gattttagat
     1621 ggagactcca aaaatattat tcaactgatt attaatgcat acaatagtat ccggtctaaa
     1681 gtggagttgt cagtctggga tcagcctgag gatcttaatc tcttctttac tgctacctgc
     1741 caagatgggg tatcctatcc tggtcagagg aagtgtgagg gtctgaagat tggggacacg
     1801 gcatcttttg aagtatcatt ggaggcccga agctgtccca gcagacacac ggagcatgtg
     1861 tttgccctgc ggccggtggg attccgggac agcctggagg tgggggtcac ctacaactgc
     1921 acgtgcggct gcagcgtggg gctggaaccc aacagcgcca ggtgcaacgg gagcgggacc
     1981 tatgtctgcg gcctgtgtga gtgcagcccc ggctacctgg gcaccaggtg cgagtgccag
     2041 gatggggaga accagagcgt gtaccagaac ctgtgccggg aggcagaggg caagccactg
     2101 tgcagcgggc gtggggactg cagctgcaac cagtgctcct gcttcgagag cgagtttggc
     2161 aagatctatg ggcctttctg tgagtgcgac aacttctcct gtgccaggaa caagggagtc
     2221 ctctgctcag gccatggcga gtgtcactgc ggggaatgca agtgccatgc aggttacatc
     2281 ggggacaact gtaactgctc gacagacatc agcacatgcc ggggcagaga tggccagatc
     2341 tgcagcgagc gtgggcactg tctctgtggg cagtgccaat gcacggagcc gggggccttt
     2401 ggggagatgt gtgagaagtg ccccacctgc ccggatgcat gcagcaccaa gagagattgc
     2461 gtcgagtgcc tgctgctcca ctctgggaaa cctgacaacc agacctgcca cagcctatgc
     2521 agggatgagg tgatcacatg ggtggacacc atcgtgaaag atgaccagga ggctgtgcta
     2581 tgtttctaca aaaccgccaa ggactgcgtc atgatgttca cctatgtgga gctccccagt
     2641 gggaagtcca acctgaccgt cctcagggag ccagagtgtg gaaacacccc caacgccatg
     2701 accatcctcc tggctgtggt cggtagcatc ctccttgttg ggcttgcact cctggctatc
     2761 tggaagctgc ttgtcaccat ccacgaccgg agggagtttg caaagtttca gagcgagcga
     2821 tccagggccc gctatgaaat ggcttcaaat ccattataca gaaagcctcg aattctgcag
     2881 tcgacggtac cgcgggcccg ggatccaccg gtcatggtga gcaagggcga ggagctgttc
     2941 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     3001 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     3061 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     3121 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     3181 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     3241 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     3301 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     3361 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     3421 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     3481 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     3541 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3601 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
     3661 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3721 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3781 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3841 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     3901 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3961 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4021 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4081 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4141 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4201 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4261 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4321 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4381 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4441 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     4501 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4561 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4621 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4681 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4741 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4801 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4861 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4921 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4981 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5041 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5101 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5161 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5221 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5281 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5341 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     5401 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5461 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5521 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5581 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5641 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5701 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5761 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5821 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5881 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     5941 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     6001 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     6061 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     6121 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6181 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6241 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6301 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6361 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6421 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6481 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6541 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6601 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6661 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6721 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6781 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6841 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6901 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6961 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     7021 gagttcc
//
