LOCUS       sequence-484519-        7080 bp    DNA     circular SYN 27-AUG-2025
DEFINITION  Encodes human integrin beta-6 chimera containing integrin beta-1 
            cytoplasmic domain and C-terminal EGFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7080)
  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 7080)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 27, 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-6-beta-1-EGFP
FEATURES             Location/Qualifiers
     source          1..7080
                     /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      3723..3732
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3729..3731,3732..3734,3735..4448)
                     /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(3729..3731,3732..3734,3735..4445)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3774..3795)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4035..4054)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4382..4403
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4571..4692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4608..4627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4662..4681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4699..5154)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4836..4857)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5048..5067
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5181..5285
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5287..5644
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5287..5478
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5310..5330)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5495..5630
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5679..6473
                     /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             5679..6473
                     /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(5733..5752)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6343..6362
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6661..6680)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6705..6752
                     /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 gatccattgt gctctaaagg caagaactga aacgaatggg gattgaactg ctttgcctgt
     1381 tctttctatt tctaggaagg aatgatcacg tacaaggtgg ctgtgccctg ggaggtgcag
     1441 aaacctgtga agactgcctg cttattggac ctcagtgtgc ctggtgtgct caggagaatt
     1501 ttactcatcc atctggagtt ggcgaaaggt gtgatacccc agcaaacctt ttagctaaag
     1561 gatgtcaatt aaacttcatc gaaaaccctg tctcccaagt agaaatactt aaaaataagc
     1621 ctctcagtgt aggcagacag aaaaatagtt ctgacattgt tcagattgcg cctcaaagct
     1681 tgatccttaa gttgagacca ggtggtgcgc agactctgca ggtgcatgtc cgccagactg
     1741 aggactaccc ggtggatttg tattacctca tggacctctc cgcctccatg gatgacgacc
     1801 tcaacacaat aaaggagctg ggctcccggc tttccaaaga gatgtctaaa ttaaccagca
     1861 actttagact gggcttcgga tcttttgtgg aaaaacctgt atcccctttc gtgaaaacaa
     1921 caccagaaga aattgccaac ccttgcagta gtattccata cttctgttta cctacatttg
     1981 gattcaagca cattttgcca ttgacaaatg atgctgaaag attcaatgaa attgtgaaga
     2041 atcagaaaat ttctgctaat attgacacac ccgaaggtgg atttgatgca attatgcaag
     2101 ctgctgtgtg taaggaaaaa attggctggc ggaatgactc cctccacctc ctggtctttg
     2161 tgagtgatgc tgattctcat tttggaatgg acagcaaact agcaggcatc gtcattccta
     2221 atgacgggct ctgtcacttg gacagcaaga atgaatactc catgtcaact gtcttggaat
     2281 atccaacaat tggacaactc attgataaac tggtacaaaa caacgtgtta ttgatcttcg
     2341 ctgtaaccca agaacaagtt catttatatg agaattacgc aaaacttatt cctggagcta
     2401 cagtaggtct acttcagaag gactccggaa acattctcca gctgatcatc tcagcttatg
     2461 aagaactgcg gtctgaggtg gaactggaag tattaggaga cactgaagga ctcaacttgt
     2521 catttacagc catctgtaac aacggtaccc tcttccaaca ccaaaagaaa tgctctcaca
     2581 tgaaagtggg agacacagct tccttcagcg tgactgtgaa tatcccacac tgcgagagaa
     2641 gaagcaggca cattatcata aagcctgtgg ggctggggga tgccctggaa ttacttgtca
     2701 gcccagaatg caactgcgac tgtcagaaag aagtggaagt gaacagctcc aaatgtcacc
     2761 acgggaacgg ctctttccag tgtggggtgt gtgcctgcca ccctggccac atggggcctc
     2821 gctgtgagtg tggcgaggac atgctgagca cagattcctg caaggaggcc ccagatcatc
     2881 cctcctgcag cggaaggggt gactgctact gtgggcagtg tatctgccac ttgtctccct
     2941 atggaaacat ttatgggcct tattgccagt gtgacaattt ctcctgcgtg agacacaaag
     3001 ggctgctctg cggaggtaac ggcgactgtg actgtggtga atgtgtgtgc aggagcggct
     3061 ggactggcga gtactgcaac tgcaccacca gcacggactc ctgcgtctct gaagatggag
     3121 tgctctgcag cgggcgcggg gactgtgttt gtggcaagtg tgtttgcaca aaccctggag
     3181 cctcaggacc aacctgtgaa cgatgtccta cctgtggtga cccctgtaac tctaaacgga
     3241 gctgcattga gtgccacctg tcagcagctg gccaagcccg agaagaatgt gtggacaagt
     3301 gcaaactagc tggtgcgacc atcagtgaag aagaagattt ctcaaaggat ggttctgttt
     3361 cctgctctct gcaaggagaa aatgaatgtc ttattacatt cctaataact acagataatg
     3421 aggggaaaac catcattcac agcatcaatg aaaaagattg tccgaagcct ccaaacattc
     3481 ccatgatcat gttaggggtt tccctggcta ttcttctcat cggggttgtc ctactgtgca
     3541 tctggaagct tttaatgata attcatgaca gaagggagtt tgctaaattt gaaaaggaga
     3601 aaatgaatgc caaatgggac acgggtgaaa atcctattta taagagtgcc gtaacaactg
     3661 tggtcaatcc gaagtatgag ggaaaatcta gagcggccgc caccgcgggc ccgggatcca
     3721 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     3781 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     3841 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     3901 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     3961 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     4021 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     4081 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     4141 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     4201 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     4261 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     4321 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     4381 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     4441 acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     4501 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     4561 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4621 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4681 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4741 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4801 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4861 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4921 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4981 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     5041 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     5101 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     5161 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5221 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5281 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5341 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5401 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5461 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5521 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5581 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5641 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5701 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5761 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5821 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     5881 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5941 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     6001 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     6061 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     6121 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6181 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     6241 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6301 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6361 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6421 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6481 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6541 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6601 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     6661 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     6721 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     6781 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     6841 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     6901 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6961 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7021 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
