LOCUS       sequence-484520-        7051 bp    DNA     circular SYN 27-AUG-2025
DEFINITION  Encodes wildtype human integrin beta-3 and C-terminal EBFP2 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7051)
  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 7051)
  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-3-EBFP2
FEATURES             Location/Qualifiers
     source          1..7051
                     /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      3695..3704
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3701..3703,3704..3706,3707..4420)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(3746..3767)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    4542..4663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4579..4598)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4633..4652
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4670..5125)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4807..4828)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5019..5038
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5152..5256
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5258..5615
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5258..5449
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5281..5301)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5466..5601
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5650..6444
                     /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             5650..6444
                     /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(5704..5723)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6314..6333
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6632..6651)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6676..6723
                     /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 gatgcgagcg cggccgcggc cccggccgct ctgggcgact gtgctggcgc tgggggcgct
     1381 ggcgggcgtt ggcgtaggag ggcccaacat ctgtaccacg cgaggtgtga gctcctgcca
     1441 gcagtgcctg gctgtgagcc ccatgtgtgc ctggtgctct gatgaggccc tgcctctggg
     1501 ctcacctcgc tgtgacctga aggagaatct gctgaaggat aactgtgccc cagaatccat
     1561 cgagttccca gtgagtgagg cccgagtact agaggacagg cccctcagcg acaagggctc
     1621 tggagacagc tcccaggtca ctcaagtcag tccccagagg attgcactcc ggctccggcc
     1681 agatgattcg aagaatttct ccatccaagt gcggcaggtg gaggattacc ctgtggacat
     1741 ctactacttg atggacctgt cttactccat gaaggatgat ctgtggagca tccagaacct
     1801 gggtaccaag ctggccaccc agatgcgaaa gctcaccagt aacctgcgga ttggcttcgg
     1861 ggcatttgtg gacaagcctg tgtcaccata catgtatatc tccccaccag aggccctcga
     1921 aaacccctgc tatgatatga agaccacctg cttgcccatg tttggctaca aacacgtgct
     1981 gacgctaact gaccaggtga cccgcttcaa tgaggaagtg aagaagcaga gtgtgtcacg
     2041 gaaccgagat gccccagagg gtggctttga tgccatcatg caggctacag tctgtgatga
     2101 aaagattggc tggaggaatg atgcatccca cttgctggtg tttaccactg atgccaagac
     2161 tcatatagca ttggacggaa ggctggcagg cattgtccag cctaatgacg ggcagtgtca
     2221 tgttggtagt gacaatcatt actctgcctc cactaccatg gattatccct ctttggggct
     2281 gatgactgag aagctatccc agaaaaacat caatttgatc tttgcagtga ctgaaaatgt
     2341 agtcaatctc tatcagaact atagtgagct catcccaggg accacagttg gggttctgtc
     2401 catggattcc agcaatgtcc tccagctcat tgttgatgct tatgggaaaa tccgttctaa
     2461 agtcgagctg gaagtgcgtg acctccctga agagttgtct ctatccttca atgccacctg
     2521 cctcaacaat gaggtcatcc ctggcctcaa gtcttgtatg ggactcaaga ttggagacac
     2581 ggtgagcttc agcattgagg ccaaggtgcg aggctgtccc caggagaagg agaagtcctt
     2641 taccataaag cccgtgggct tcaaggacag cctgatcgtc caggtcacct ttgattgtga
     2701 ctgtgcctgc caggcccaag ctgaacctaa tagccatcgc tgcaacaatg gcaatgggac
     2761 ctttgagtgt ggggtatgcc gttgtgggcc tggctggctg ggatcccagt gtgagtgctc
     2821 agaggaggac tatcgccctt cccagcagga cgaatgcagc ccccgggagg gtcagcccgt
     2881 ctgcagccag cggggcgagt gcctctgtgg tcaatgtgtc tgccacagca gtgactttgg
     2941 caagatcacg ggcaagtact gcgagtgtga cgacttctcc tgtgtccgct acaaggggga
     3001 gatgtgctca ggccatggcc agtgcagctg tggggactgc ctgtgtgact ccgactggac
     3061 cggctactac tgcaactgta ccacgcgtac tgacacctgc atgtccagca atgggctgct
     3121 gtgcagcggc cgcggcaagt gtgaatgtgg cagctgtgtc tgtatccagc cgggctccta
     3181 tggggacacc tgtgagaagt gccccacctg cccagatgcc tgcaccttta agaaagaatg
     3241 tgtggagtgt aagaagtttg accggggagc cctacatgac gaaaatacct gcaaccgtta
     3301 ctgccgtgac gagattgagt cagtgaaaga gcttaaggac actggcaagg atgcagtgaa
     3361 ttgtacctat aagaatgagg atgactgtgt cgtcagattc cagtactatg aagattctag
     3421 tggaaagtcc atcctgtatg tggtagaaga gccagagtgt cccaagggcc ctgacatcct
     3481 ggtggtcctg ctctcagtga tgggggccat tctgctcatt ggccttgccg ccctgctcat
     3541 ctggaaactc ctcatcacca tccacgaccg aaaagaattc gctaaatttg aggaagaacg
     3601 cgccagagca aaatgggaca cagccaacaa cccactgtat aaagaggcca cgtctacctt
     3661 caccaatatc acgtaccggg gcacttcacc ggtcgccacc atggtgagca agggcgagga
     3721 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3781 gttcagcgtg aggggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     3841 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgagcca
     3901 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3961 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaccta
     4021 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     4081 gggcgtcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa
     4141 cagccacaac atctatatca tggccgtcaa gcagaagaac ggcatcaagg tgaacttcaa
     4201 gatccgccac aacgtggagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     4261 ccccatcggc gacggccccg tgctgctgcc cgacagccac tacctgagca cccagtccgt
     4321 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tccgcaccgc
     4381 cgccgggatc actctcggca tggacgagct gtacaagtaa gcggccgcga ctctagatca
     4441 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     4501 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     4561 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4621 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     4681 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4741 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4801 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4861 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4921 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4981 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5041 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5101 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5161 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5221 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     5281 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5341 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     5401 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5461 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5521 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     5581 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     5641 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5701 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5761 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5821 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5881 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5941 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6001 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6061 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6121 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     6181 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6241 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6301 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6361 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6421 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     6481 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     6541 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     6601 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     6661 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     6721 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     6781 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     6841 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     6901 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6961 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7021 cagaccccgt agaaaagatc aaaggatctt c
//
