LOCUS       sequence-199532-        5823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription activator (transactivation domain of viral VP16 
            protein) that is constitutively recruited lacO arrays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5823)
  AUTHORS   Rademacher A, Erdel F, Trojanowski J, Schumacher S, Rippe K
  TITLE     Real-time observation of light-controlled transcription in living 
            cells.
  JOURNAL   J Cell Sci. 2017 Dec 15;130(24):4213-4224. doi: 10.1242/jcs.205534. 
            Epub 2017 Nov 9.
  PUBMED    29122982
REFERENCE   2  (bases 1 to 5823)
  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: 
            "10.1242/jcs.205534"; journalName: "J Cell Sci"; date: "2017-12-15- 
            15"; volume: "130"; issue: "24"; pages: "4213-4224"
COMMENT     Sequence Label: ptagBFP-LacI-NLS-VP16
FEATURES             Location/Qualifiers
     source          1..5823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     501..520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        733..1062
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      913..1048
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1107..1805
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(1843..1862)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2922..2942
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3325..3446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3362..3381)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3416..3435
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3453..3908)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3590..3611)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3802..3821
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3935..4039
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4041..4398
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4041..4232
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      4249..4384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4433..5227
                     /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             4433..5227
                     /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(4487..4506)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5097..5116
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5415..5434)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5459..5506
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
       61 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      121 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
      181 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      241 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      301 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      361 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      421 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      481 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      541 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      601 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      661 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
      721 ttactgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
      781 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
      841 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
      901 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
      961 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     1021 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctagc gctaccgggg
     1081 ccgctctagc gctaccggtc gccaccatga gcgagctgat taaggagaac atgcacatga
     1141 agctgtacat ggagggcacc gtggacaacc atcacttcaa gtgcacatcc gagggcgaag
     1201 gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc
     1261 ccttcgcctt cgacatcctg gctactagct tcctctacgg cagcaagacc ttcatcaacc
     1321 acacccaggg catccccgac ttcttcaagc agtccttccc tgagggcttc acatgggaga
     1381 gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg
     1441 acggctgcct catctacaac gtcaagatca gaggggtgaa cttcacatcc aacggccctg
     1501 tgatgcagaa gaaaacactc ggctgggagg ccttcaccga gacgctgtac cccgctgacg
     1561 gcggcctgga aggcagaaac gacatggccc tgaagctcgt gggcgggagc catctgatcg
     1621 caaacatcaa gaccacatat agatccaaga aacccgctaa gaacctcaag atgcctggcg
     1681 tctactatgt ggactacaga ctggaaagaa tcaaggaggc caacaacgag acctacgtcg
     1741 agcagcacga ggtggcagtg gccagatact gcgacctccc tagcaaactg gggcacaagc
     1801 ttaattccgg actcagatcc atggtgaaac cagtaacgtt atacgatgtc gcagagtatg
     1861 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     1921 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     1981 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     2041 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     2101 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     2161 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     2221 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     2281 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     2341 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     2401 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     2461 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     2521 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     2581 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     2641 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     2701 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     2761 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
     2821 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     2881 tttcccgaag cagcctgagg gggcgtcgac tgggtacccc tcctaagaag aagaggaagg
     2941 ttgcccccac catggataaa aagaagagaa aggccccccc gaccgatgtc agcctggggg
     3001 acgagctcca cttagacggc gaggacgtgg cgatggcgca tgccgacgcg ctagacgatt
     3061 tcgatctgga catgttgggg gacggggatt ccccgggtcc gggatttacc ccccacgact
     3121 ccgcccccta cggcgctctg gatatggccg acttcgagtt tgagcagatg tttaccgatg
     3181 cccctctaga cggggggatc caccggatct agataactga tcataatcag ccataccaca
     3241 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3301 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3361 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3421 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     3481 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3541 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3601 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3661 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3721 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3781 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3841 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3901 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3961 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4021 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4081 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4141 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4201 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4261 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4321 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4381 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4441 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4501 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4561 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4621 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4681 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4741 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4801 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4861 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4921 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4981 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5041 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5101 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5161 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5221 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5281 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5341 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     5401 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5461 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5521 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5581 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5641 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5701 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5761 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5821 atc
//
