LOCUS       sequence-266008-        7153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Beta1-GFP integrin for normal cell transfection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7153)
  AUTHORS   Parsons M, Messent AJ, Humphries JD, Deakin NO, Humphries MJ
  TITLE     Quantification of integrin receptor agonism by fluorescence lifetime
            imaging.
  JOURNAL   J Cell Sci. 2008 Feb 1;121(Pt 3):265-71. doi: 10.1242/jcs.018440.
  PUBMED    18216331
REFERENCE   2  (bases 1 to 7153)
  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: "J Cell 
            Sci."; date: "2008-02-1"; volume: "121(Pt 3):265-71. doi"; pages: " 
            10.1242/jcs.018440"
COMMENT     Sequence Label: Beta1-GFP in pHcgreen
FEATURES             Location/Qualifiers
     source          1..7153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3093..3102
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3099..3101,3102..3104,3105..3818)
                     /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(3099..3101,3102..3104,3105..3815)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3144..3165)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3405..3424)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3752..3773
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3941..4062
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3978..3997)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4032..4051
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4069..4524)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4206..4227)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4418..4437
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4551..4655
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4657..5014
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4657..4848
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4680..4700)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4865..5000
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5049..5843
                     /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             5049..5843
                     /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(5103..5122)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5713..5732
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6031..6050)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6075..6122
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6451..7039
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6940..6959
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc atgaatttac
      661 aaccaatttt ctggattgga ctgatcagtt cagtttgctg tgtgtttgct caaacagatg
      721 aaaatagatg tttaaaagca aatgccaaat catgtggaga atgtatacaa gcagggccaa
      781 attgtgggtg gtgcacaaat tcaacatttt tacaggaagg aatgcctact tctgcacgat
      841 gtgatgattt agaagcctta aaaaagaagg gttgccctcc agatgacata gaaaatccca
      901 gaggctccaa agatataaag aaaaataaaa atgtaaccaa ccgtagcaaa ggaacagcag
      961 agaagctcaa gccagaggat attactcaga tccaaccaca gcagttggtt ttgcgattaa
     1021 gatcagggga gccacagaca tttacattaa aattcaagag agctgaagac tatcccattg
     1081 acctctacta ccttatggac ctgtcttact caatgaaaga cgatttggag aatgtaaaaa
     1141 gtcttggaac agatctgatg aatgaaatga ggaggattac ttcggacttc agaattggat
     1201 ttggctcatt tgtggaaaag actgtgatgc cttacattag cacaacacca gctaagctca
     1261 ggaacccttg cacaagtgaa cagaactgca ccagcccatt tagctacaaa aatgtgctca
     1321 gtcttactaa taaaggagaa gtatttaatg aacttgttgg aaaacagcgc atatctggaa
     1381 atttggattc tccagaaggt ggtttcgatg ccatcatgca agttgcagtt tgtggatcac
     1441 tgattggctg gaggaatgtt acacggctgc tggtgttttc cacagatgcc gggtttcact
     1501 ttgctggaga tgggaaactt ggtggcattg ttttaccaaa tgatggacaa tgtcacctgg
     1561 aaaataatat gtacacaatg agccattatt atgattatcc ttctattgct caccttgtcc
     1621 agaaactgag tgaaaataat attcagacaa tttttgcagt tactgaagaa tttcagcctg
     1681 tttacaagga gctgaaaaac ttgatcccta agtcagcagt aggaacatta tctgcaaatt
     1741 ctagcaatgt aattcagttg atcattgatg catacaattc cctttcctca gaagtcattt
     1801 tggaaaacgg caaattgtca gaaggagtaa caataagtta caaatcttac tgcaagaacg
     1861 gggtgaatgg aacaggggaa aatggaagaa aatgttccaa tatttccatt ggagatgagg
     1921 ttcaatttga aattagcata acttcaaata agtgtccaaa aaaggattct gacagcttta
     1981 aaattaggcc tctgggcttt acggaggaag tagaggttat tcttcagtac atctgtgaat
     2041 gtgaatgcca aagcgaaggc atccctgaaa gtcccaagtg tcatgaagga aatgggacat
     2101 ttgagtgtgg cgcgtgcagg tgcaatgaag ggcgtgttgg tagacattgt gaatgcagca
     2161 cagatgaagt taacagtgaa gacatggatg cttactgcag gaaagaaaac agttcagaaa
     2221 tctgcagtaa caatggagag tgcgtctgcg gacagtgtgt ttgtaggaag agggataata
     2281 caaatgaaat ttattctggc aaattctgcg agtgtgataa tttcaactgt gatagatcca
     2341 atggcttaat ttgtggagga aatggtgttt gcaagtgtcg tgtgtgtgag tgcaacccca
     2401 actacactgg cagtgcatgt gactgttctt tggatactag tacttgtgaa gccagcaacg
     2461 gacagatctg caatggccgg ggcatctgcg agtgtggtgt ctgtaagtgt acagatccga
     2521 agtttcaagg gcaaacgtgt gagatgtgtc agacctgcct tggtgtctgt gctgagcata
     2581 aagaatgtgt tcagtgcaga gccttcaata aaggagaaaa gaaagacaca tgcacacagg
     2641 aatgttccta ttttaacatt accaaggtag aaagtcggga caaattaccc cagccggtcc
     2701 aacctgatcc tgtgtcccat tgtaaggaga aggatgttga cgactgttgg ttctatttta
     2761 cgtattcagt gaatgggaac aacgaggtca tggttcatgt tgtggagaat ccagagtgtc
     2821 ccactggtcc agacatcatt ccaattgtag ctggtgtggt tgctggaatt gttcttattg
     2881 gccttgcatt actgctgata tggaagcttt taatgataat tcatgacaga agggagtttg
     2941 ctaaatttga aaaggagaaa atgaatgcca aatgggacac gggtgaaaat cctatttata
     3001 agagtgccgt aacaactgtg gtcaatccga agtatgaggg aaaaggaggg gggggggccc
     3061 ggaggcgggg ggaggcgggg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc
     3121 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     3181 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     3241 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     3301 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     3361 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     3421 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3481 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     3541 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     3601 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     3661 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     3721 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     3781 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat
     3841 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3901 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3961 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4021 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     4081 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     4141 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     4201 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     4261 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     4321 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     4381 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     4441 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4501 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4561 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4621 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     4681 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4741 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4801 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4861 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4921 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4981 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     5041 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5101 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     5161 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     5221 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     5281 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5341 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     5401 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5461 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5521 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     5581 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5641 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5701 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5761 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5821 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5881 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5941 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     6001 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     6061 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     6121 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     6181 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     6241 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     6301 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6361 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6421 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6481 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6541 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6601 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6661 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6721 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6781 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6841 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6901 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6961 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7021 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7081 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     7141 taccgccatg cat
//
