LOCUS       sequence-486257-        7186 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Encodes human integrin beta-1 chimera containing integrin beta-6 
            cytoplasmic domain and C-terminal EGFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7186)
  AUTHORS   Lee WN, Li J, Chen NP, Yu CH
  TITLE     Kindlin Regulates Mechanosensitive Activation and Adhesion Assembly 
            of Integrin beta6.
  JOURNAL   Adv Sci (Weinh). 2025 Jun 10:e01078. doi: 10.1002/advs.202501078.
  PUBMED    40492509
REFERENCE   2  (bases 1 to 7186)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Adv Sci 
            (Weinh). 2025 Jun 10:e01078. doi: 10.1002/advs.202501078."
COMMENT     Sequence Label: Integrin beta-1-beta-6-EGFP
FEATURES             Location/Qualifiers
     source          1..7186
                     /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      3829..3838
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3835..3837,3838..3840,3841..4554)
                     /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(3835..3837,3838..3840,3841..4551)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3880..3901)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4141..4160)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4488..4509
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4677..4798
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4714..4733)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4768..4787
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4805..5260)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4942..4963)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5154..5173
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5287..5391
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5393..5750
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5393..5584
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5416..5436)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5601..5736
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5785..6579
                     /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             5785..6579
                     /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(5839..5858)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6449..6468
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6767..6786)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6811..6858
                     /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 cgaattctgc agtcgacggt accatgaatt tacaaccaat tttctggatt
     1381 ggactgatca gttcagtttg ctgtgtgttt gctcaaacag atgaaaatag atgtttaaaa
     1441 gcaaatgcca aatcatgtgg agaatgtata caagcagggc caaattgtgg gtggtgcaca
     1501 aattcaacat ttttacagga aggaatgcct acttctgcac gatgtgatga tttagaagcc
     1561 ttaaaaaaga agggttgccc tccagatgac atagaaaatc ccagaggctc caaagatata
     1621 aagaaaaata aaaatgtaac caaccgtagc aaaggaacag cagagaagct caagccagag
     1681 gatattactc agatccaacc acagcagttg gttttgcgat taagatcagg ggagccacag
     1741 acatttacat taaaattcaa gagagctgaa gactatccca ttgacctcta ctaccttatg
     1801 gacctgtctt actcaatgaa agacgatttg gagaatgtaa aaagtcttgg aacagatctg
     1861 atgaatgaaa tgaggaggat tacttcggac ttcagaattg gatttggctc atttgtggaa
     1921 aagactgtga tgccttacat tagcacaaca ccagctaagc tcaggaaccc ttgcacaagt
     1981 gaacagaact gcaccagccc atttagctac aaaaatgtgc tcagtcttac taataaagga
     2041 gaagtattta atgaacttgt tggaaaacag cgcatatctg gaaatttgga ttctccagaa
     2101 ggtggtttcg atgccatcat gcaagttgca gtttgtggat cactgattgg ctggaggaat
     2161 gttacacggc tgctggtgtt ttccacagat gccgggtttc actttgctgg agatgggaaa
     2221 cttggtggca ttgttttacc aaatgatgga caatgtcacc tggaaaataa tatgtacaca
     2281 atgagccatt attatgatta tccttctatt gctcaccttg tccagaaact gagtgaaaat
     2341 aatattcaga caatttttgc agttactgaa gaatttcagc ctgtttacaa ggagctgaaa
     2401 aacttgatcc ctaagtcagc agtaggaaca ttatctgcaa attctagcaa tgtaattcag
     2461 ttgatcattg atgcatacaa ttccctttcc tcagaagtca ttttggaaaa cggcaaattg
     2521 tcagaaggag taacaataag ttacaaatct tactgcaaga acggggtgaa tggaacaggg
     2581 gaaaatggaa gaaaatgttc caatatttcc attggagatg aggttcaatt tgaaattagc
     2641 ataacttcaa ataagtgtcc aaaaaaggat tctgacagct ttaaaattag gcctctgggc
     2701 tttacggagg aagtagaggt tattcttcag tacatctgtg aatgtgaatg ccaaagcgaa
     2761 ggcatccctg aaagtcccaa gtgtcatgaa ggaaatggga catttgagtg tggcgcgtgc
     2821 aggtgcaatg aagggcgtgt tggtagacat tgtgaatgca gcacagatga agttaacagt
     2881 gaagacatgg atgcttactg caggaaagaa aacagttcag aaatctgcag taacaatgga
     2941 gagtgcgtct gcggacagtg tgtttgtagg aagagggata atacaaatga aatttattct
     3001 ggcaaattct gcgagtgtga taatttcaac tgtgatagat ccaatggctt aatttgtgga
     3061 ggaaatggtg tttgcaagtg tcgtgtgtgt gagtgcaacc ccaactacac tggcagtgca
     3121 tgtgactgtt ctttggatac tagtacttgt gaagccagca acggacagat ctgcaatggc
     3181 cggggcatct gcgagtgtgg tgtctgtaag tgtacagatc cgaagtttca agggcaaacg
     3241 tgtgagatgt gtcagacctg ccttggtgtc tgtgctgagc ataaagaatg tgttcagtgc
     3301 agagccttca ataaaggaga aaagaaagac acatgcacac aggaatgttc ctattttaac
     3361 attaccaagg tagaaagtcg ggacaaatta ccccagccgg tccaacctga tcctgtgtcc
     3421 cattgtaagg agaaggatgt tgacgactgt tggttctatt ttacgtattc agtgaatggg
     3481 aacaacgagg tcatggttca tgttgtggag aatccagagt gtcccactgg tccagacatc
     3541 attccaattg tagctggtgt ggttgctgga attgttctta ttggccttgc attactgctg
     3601 atatggaagc tactggtgtc atttcatgat cgtaaagaag ttgccaaatt tgaagcagaa
     3661 cgatcaaaag ccaagtggca aacgggaacc aatccactct acagaggatc cacaagtact
     3721 tttaaaaatg taacttataa acacagggaa aaacaaaagg tagacctttc cacagattgc
     3781 ggaggggggg gggcccggag gcggggggag gcgggggatc caccggtcgc caccatggtg
     3841 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     3901 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     3961 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     4021 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     4081 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     4141 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     4201 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     4261 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     4321 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     4381 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     4441 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     4501 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     4561 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     4621 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     4681 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4741 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     4801 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4861 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4921 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4981 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     5041 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     5101 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     5161 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5221 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     5281 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5341 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     5401 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5461 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5521 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5581 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5641 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     5701 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     5761 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5821 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5881 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5941 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     6001 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     6061 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     6121 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     6181 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     6241 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     6301 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     6361 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     6421 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6481 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6541 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     6601 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     6661 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     6721 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     6781 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     6841 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     6901 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     6961 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     7021 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7081 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7141 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
