LOCUS       sequence-321678-        6907 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains integrin beta 6 residues 1–474 with I270C mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6907)
  AUTHORS   Dong X, Hudson NE, Lu C, Springer TA
  TITLE     Structural determinants of integrin beta-subunit specificity for 
            latent TGF-beta.
  JOURNAL   Nat Struct Mol Biol. 2014 Dec;21(12):1091-6. doi: 10.1038/nsmb.2905.
            Epub 2014 Nov 10.
  PUBMED    25383667
REFERENCE   2  (bases 1 to 6907)
  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/nsmb.2905";
            journalName: "Nat Struct Mol Biol"; date: "2014-12"; volume: "21"; 
            issue: "12"; pages: "1091-6"
COMMENT     Sequence Label: Integrin-beta-6 Headpiece I270C
FEATURES             Location/Qualifiers
     source          1..6907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        43..346
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        347..550
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     501..521
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      589..598
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2203..2220
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    join(2248..2822,2823..2825,2826..2834)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2415..2432)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2642..2661
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2835..2837,2838..2840,2841..3554)
                     /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(2835..2837,2838..2840,2841..3551)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2880..2901)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3141..3160)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3488..3509
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3677..3798
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3714..3733)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3768..3787
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3805..4260)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3942..3963)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4154..4173
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4287..4391
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4393..4750
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4393..4584
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4416..4436)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4601..4736
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4785..5579
                     /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             4785..5579
                     /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(4839..4858)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5449..5468
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5767..5786)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5811..5858
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6187..6775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6676..6695
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
       61 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      121 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      181 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      241 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      301 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      361 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      421 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      481 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      541 aagcagagct ggtttagtga accgtcagat ccgctagcct cgacgtacgc caccatggat
      601 atgagagttc ctgctcaatt gctggggttg ctcttgctct ggttcagtgg ggtgttgggg
      661 cacgtacaag gtggctgtgc cctgggaggt gcagaaacct gtgaagactg cctgcttatt
      721 ggacctcagt gtgcctggtg tgctcaggag aattttactc atccatctgg agttggcgaa
      781 aggtgtgata ccccagcaaa ccttttagct aaaggatgtc aattaaactt catcgaaaac
      841 cctgtctccc aagtagaaat acttaaaaat aagcctctca gtgtaggcag acagaaaaat
      901 agttctgaca ttgttcagat tgcgcctcaa agcttgatcc ttaagttgag accaggtggt
      961 gcgcagactc tgcaggtgca tgtccgccag actgaggact acccggtgga tttgtattac
     1021 ctcatggacc tctccgcctc catggatgac gacctcaaca caataaagga gctgggctcc
     1081 cggctttcca aagagatgtc taaattaacc agcaacttta gactgggctt cggatctttt
     1141 gtggaaaaac ctgtatcccc tttcgtgaaa acaacaccag aagaaattgc caacccttgc
     1201 agtagtattc catacttctg tttacctaca tttggattca agcacatttt gccattgaca
     1261 aatgatgctg aaagattcaa tgaaattgtg aagaatcaga aaatttctgc taatattgac
     1321 acacccgaag gtggatttga tgcaattatg caagctgctg tgtgtaagga aaaaattggc
     1381 tggcggaatg actccctcca cctcctggtc tttgtgagtg atgctgattc tcattttgga
     1441 atggacagca aactagcagg catcgtctgt cctaatgacg ggctctgtca cttggacagc
     1501 aagaatgaat actccatgtc aactgtcttg gaatatccaa caattggaca actcattgat
     1561 aaactggtac aaaacaacgt gttattgatc ttcgctgtaa cccaagaaca agttcattta
     1621 tatgagaatt acgcaaaact tattcctgga gctacagtag gtctacttca gaaggactcc
     1681 ggaaacattc tccagctgat catctcagct tatgaagaac tgcggtctga ggtggaactg
     1741 gaagtattag gagacactga aggactcaac ttgtcattta cagccatctg taacaacggt
     1801 accctcttcc aacaccaaaa gaaatgctct cacatgaaag tgggagacac agcttccttc
     1861 agcgtgactg tgaatatccc acactgcgag agaagaagca ggcacattat cataaagcct
     1921 gtggggctgg gggatgccct ggaattactt gtcagcccag aatgcaactg cgactgtcag
     1981 aaagaagtgg aagtgaacag ctccaaatgt caccacggga acggctcttt ccagtgtggg
     2041 gtgtgtgcct gccaccctgg ccacatgggg cctcgctgtg agctccaaac cctcttccaa
     2101 ggccccctcg gcgcgcagct caagaagaaa ttgcaggcgc tgaagaagaa gaacgctcag
     2161 cttaagtgga aactgcaagc tctaaagaag aagctagccc agcatcacca tcaccatcac
     2221 gcgtaaacta gtggcgcgcc ggatccgccc ctctccctcc ccccccccta acgttactgg
     2281 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     2341 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     2401 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     2461 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     2521 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     2581 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     2641 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     2701 tatgggatct gatctggggc ctcggtacac atgctttaca tgtgtttagt cgaggttaaa
     2761 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     2821 atatggccac aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2881 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2941 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     3001 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     3061 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     3121 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     3181 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     3241 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     3301 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     3361 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3421 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     3481 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3541 agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     3601 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     3661 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3721 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3781 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     3841 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3901 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3961 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4021 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4081 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4141 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4201 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4261 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4321 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4381 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     4441 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4501 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4561 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4621 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4681 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4741 cttttgcaaa gatcgatcaa tagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4801 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4861 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4921 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     4981 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5041 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5101 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5161 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5221 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5281 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     5341 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5401 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5461 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5521 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5581 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5641 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5701 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     5761 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     5821 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     5881 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     5941 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     6001 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     6061 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6121 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6181 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6241 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6301 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6361 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6421 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6481 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6541 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6601 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6661 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6721 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6781 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6841 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     6901 agtaatc
//
