LOCUS       sequence-406227-        7006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GFP-tagged human integrin beta-8 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7006)
  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 7006)
  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-8-GFP
FEATURES             Location/Qualifiers
     source          1..7006
                     /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      3649..3658
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3655..3657,3658..3660,3661..4374)
                     /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(3655..3657,3658..3660,3661..4371)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3700..3721)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3961..3980)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4308..4329
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4497..4618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4534..4553)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4588..4607
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4625..5080)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4762..4783)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4974..4993
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5107..5211
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5213..5570
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5213..5404
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5236..5256)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5421..5556
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5605..6399
                     /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             5605..6399
                     /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(5659..5678)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6269..6288
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6587..6606)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6631..6678
                     /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 ccaccatgtg cggctcggcc
     1321 ctggcttttt ttaccgctgc atttgtctgc ctgcaaaacg accggcgagg tcccgcctcg
     1381 ttcctctggg cagcctgggt gttttcactt gttcttggac tgggccaagg tgaagacaat
     1441 agatgtgcat cttcaaatgc agcatcctgt gccaggtgcc ttgcgctggg tccagaatgt
     1501 ggatggtgtg ttcaagagga tttcatttca ggtggatcaa gaagtgaacg ttgtgatatt
     1561 gtttccaatt taataagcaa aggctgctca gttgattcaa tagaataccc atctgtgcat
     1621 gttataatac ccactgaaaa tgaaattaat acccaggtga caccaggaga agtgtctatc
     1681 cagctgcgtc caggagccga agctaatttt atgctgaaag ttcatcctct gaagaaatat
     1741 cctgtggatc tttattatct tgttgatgtc tcagcatcaa tgcacaataa tatagaaaaa
     1801 ttaaattccg ttggaaacga tttatctaga aaaatggcat ttttctcccg tgactttcgt
     1861 cttggatttg gctcatacgt tgataaaaca gtttcaccat acattagcat ccaccccgaa
     1921 aggattcata atcaatgcag tgactacaat ttagactgca tgcctcccca tggatacatc
     1981 catgtgctgt ctttgacaga gaacatcact gagtttgaga aagcagttca tagacagaag
     2041 atctctggaa acatagatac accagaagga ggttttgacg ccatgcttca ggcagctgtc
     2101 tgtgaaagtc atatcggatg gcgaaaagag gctaaaagat tgctgctggt gatgacagat
     2161 cagacgtctc atctcgctct tgatagcaaa ttggcaggca tagtggtgcc caatgacgga
     2221 aactgtcatc tgaaaaacaa cgtctacgtc aaatcgacaa ccatggaaca cccctcacta
     2281 ggccaacttt cagagaaatt aatagacaac aacattaatg tcatctttgc agttcaagga
     2341 aaacaatttc attggtataa ggatcttcta cccctcttgc caggcaccat tgctggtgaa
     2401 atagaatcaa aggctgcaaa cctcaataat ttggtagtgg aagcctatca gaagctcatt
     2461 tcagaagtga aagttcaggt ggaaaaccag gtacaaggca tctattttaa cattaccgcc
     2521 atctgtccag atgggtccag aaagccaggc atggaaggat gcagaaacgt gacgagcaat
     2581 gatgaagttc ttttcaatgt aacagttaca atgaaaaaat gtgatgtcac aggaggaaaa
     2641 aactatgcaa taatcaaacc tattggtttt aatgaaaccg ctaaaattca tatacacaga
     2701 aactgcagct gtcagtgtga ggacaacaga ggacctaaag gaaagtgtgt agatgaaact
     2761 tttctagatt ccaagtgttt ccagtgtgat gagaataaat gtcattttga tgaagatcag
     2821 ttttcttctg agagttgcaa gtcacacaag gatcagcctg tttgcagtgg tcgaggagtt
     2881 tgtgtttgtg ggaaatgttc atgtcacaaa attaagcttg gaaaagtgta tggaaaatac
     2941 tgtgaaaagg atgacttttc ttgtccatat caccatggaa atctgtgtgc tgggcatgga
     3001 gagtgtgaag caggcagatg ccaatgcttc agtggctggg aaggtgatcg atgccagtgc
     3061 ccttcagcag cagcccagca ctgtgtcaat tcaaagggcc aagtgtgcag tggaagaggc
     3121 acgtgtgtgt gtggaaggtg tgagtgcacc gatcccagga gcatcggccg cttctgtgaa
     3181 cactgcccca cctgttatac agcctgcaag gaaaactgga attgtatgca atgccttcac
     3241 cctcacaatt tgtctcaggc tatacttgat cagtgcaaaa cctcatgtgc tctcatggaa
     3301 caacagcatt atgtcgacca aacttcagaa tgtttctcca gcccaagcta cttgagaata
     3361 tttttcatca ttttcatagt tacattcttg attgggttgc ttaaagtcct gatcattaga
     3421 caggtgatac tacaatggaa tagtaataaa attaagtcct catcagatta cagagtgtca
     3481 gcctcaaaaa aggataagtt gattctgcaa agtgtttgca caagagcagt cacctaccga
     3541 cgtgagaagc ctgaagaaat aaaaatggat atcagcaaat taaatgctca tgaaactttc
     3601 aggtgcaact tccactcgac ggtaccgcgg gcccgggatc caccggtcgc caccatggtg
     3661 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     3721 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     3781 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     3841 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     3901 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     3961 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     4021 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     4081 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     4141 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     4201 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     4261 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     4321 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     4381 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     4441 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     4501 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4561 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     4621 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4681 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4741 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4801 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4861 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4921 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4981 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5041 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     5101 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5161 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     5221 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5281 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5341 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5401 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5461 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     5521 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     5581 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5641 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5701 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5761 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     5821 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5881 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5941 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     6001 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     6061 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     6121 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     6181 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     6241 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6301 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6361 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     6421 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     6481 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     6541 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     6601 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     6661 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     6721 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     6781 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     6841 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6901 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6961 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
