LOCUS       sequence-201206-        5358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription activator (transactivation domain of viral VP16 
            protein) that is constitutively recruited tetO arrays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5358)
  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 5358)
  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-TetR-NLS-VP16
FEATURES             Location/Qualifiers
     source          1..5358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     496..515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        728..1057
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      908..1043
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1102..1800
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             1807..2430
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MVSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1883..1902)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             2452..2472
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2855..2976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2892..2911)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2946..2965
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2983..3438)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3120..3141)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3332..3351
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3465..3569
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3571..3928
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3571..3762
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      3779..3914
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3963..4757
                     /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             3963..4757
                     /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(4017..4036)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4627..4646
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4945..4964)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4989..5036
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
       61 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      121 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      661 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattact
      721 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
      781 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
      841 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
      901 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
      961 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     1021 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctagcgctac cggggccgct
     1081 ctagcgctac cggtcgccac catgagcgag ctgattaagg agaacatgca catgaagctg
     1141 tacatggagg gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag
     1201 ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc
     1261 gccttcgaca tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc
     1321 cagggcatcc ccgacttctt caagcagtcc ttccctgagg gcttcacatg ggagagagtc
     1381 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc
     1441 tgcctcatct acaacgtcaa gatcagaggg gtgaacttca catccaacgg ccctgtgatg
     1501 cagaagaaaa cactcggctg ggaggccttc accgagacgc tgtaccccgc tgacggcggc
     1561 ctggaaggca gaaacgacat ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac
     1621 atcaagacca catatagatc caagaaaccc gctaagaacc tcaagatgcc tggcgtctac
     1681 tatgtggact acagactgga aagaatcaag gaggccaaca acgagaccta cgtcgagcag
     1741 cacgaggtgg cagtggccag atactgcgac ctccctagca aactggggca caagcttaat
     1801 tccggaatgg tgtctagatt agataaaagt aaagtgatta acagcgcatt agagctgctt
     1861 aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag
     1921 cagcctacat tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt
     1981 gagatgttag ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat
     2041 tttttacgta ataacgctaa aagttttaga tgtgctttac taagtcatcg cgatggagca
     2101 aaagtacatt taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta
     2161 gcctttttat gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgctgtg
     2221 gggcatttta ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa
     2281 agggaaacac ctactactga tagtatgccg ccattattac gacaagctat cgaattattt
     2341 gatcaccaag gtgcagagcc agccttctta ttcggccttg aattgatcat atgcggatta
     2401 gaaaaacaac ttaaatgtga aagtgggtct gaccgtcgac tgggtacccc tcctaagaag
     2461 aagaggaagg ttgcccccac catggataaa aagaagagaa aggccccccc gaccgatgtc
     2521 agcctggggg acgagctcca cttagacggc gaggacgtgg cgatggcgca tgccgacgcg
     2581 ctagacgatt tcgatctgga catgttgggg gacggggatt ccccgggtcc gggatttacc
     2641 ccccacgact ccgcccccta cggcgctctg gatatggccg acttcgagtt tgagcagatg
     2701 tttaccgatg cccctctaga cggggggatc caccggatct agataactga tcataatcag
     2761 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2821 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2881 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2941 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     3001 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3061 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3121 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3181 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3241 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3301 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3361 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3421 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3481 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3541 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3601 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3661 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3721 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3781 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3841 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3901 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3961 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4021 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4081 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4141 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4201 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4261 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4321 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4381 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4441 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4501 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4561 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4621 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4681 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4741 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4801 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4861 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4921 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4981 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5041 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5101 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5161 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5221 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5281 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5341 cgtagaaaag atcaaagg
//
