LOCUS       sequence-364097-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reverse Tet repressor labeled with GFP; binds tetO sites upon 
            doxycycline addition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5455)
  AUTHORS   Trojanowski J, Frank L, Rademacher A, Mucke N, Grigaitis P, Rippe K
  TITLE     Transcription activation is enhanced by multivalent interactions 
            independent of phase separation.
  JOURNAL   Mol Cell. 2022 May 4. pii: S1097-2765(22)00328-8. doi: 
            10.1016/j.molcel.2022.04.017.
  PUBMED    35537448
REFERENCE   2  (bases 1 to 5455)
  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: "Mol Cell. 
            2022 May 4. pii: S1097-2765(22)00328-8. doi: 
            10.1016/j.molcel.2022.04.017."
COMMENT     Sequence Label: rTetR-GFP
FEATURES             Location/Qualifiers
     source          1..5455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     170..189
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     429..450
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(1194..1397)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1223..1243)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1398..1701
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1876..2464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1956..1975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2793..2840
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2865..2884
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3072..3866)
                     /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             complement(3072..3866)
                     /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(3183..3202)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3793..3812
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3901..4258)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3915..4050
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4067..4258
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4215..4235
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4260..4364)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4391..4846
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4478..4497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4688..4709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4853..4974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4864..4883)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4918..4937
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5180..5200)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(5234..5455,1..489,490..492,493..495))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(5234..5455,1..489,490..492,493..495))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5276..5297)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
       61 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
      121 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
      181 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
      241 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
      301 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
      361 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
      421 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
      481 gcccttgctc accatgaccc gggccgatcg ggtcggcccg ccgctttcgc actttagctg
      541 tttctccagg ccacatatga ttagttccag gccgaaaagg aaggcaggtt cggctccctg
      601 ccggtcgaac agctcaattg cttgtttcag aagtgggggc atagaatcgg tggtaggtgt
      661 ctctctttcc tcttttgcta cttgatgctc ctgttcctcc aatacgcagc ccagtgtaaa
      721 gtggcccacg gcggacagag cgtacagtgc gttctccagg gagaagcctt gctgacacag
      781 gaacgcgagc tgattttcca gggtttcgta ctgtttctct gttgggcggg tgccgagatg
      841 cactttagcc ccgtcgcgat gtgagaggag agcacagcgg tatgacttgg cgttgttccg
      901 cagaaagtct tgccatgact cgccttccag ggggcaggag tgggtatgat gcctgtccag
      961 catctcgatt ggcagggcat cgagcagggc ccgcttgttc ttcacgtgcc agtacagggt
     1021 aggctgctca actcccagct tttgagcgag tttccttgtc gtcaggcctt cgataccgac
     1081 tccattgagt aattccagag cagagtttat gactttgctc ttgtccagtc tagacatggt
     1141 ggcctcgaga tctgagtccg gtagcgctag cggatctgac ggttcactaa accagctctg
     1201 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg
     1261 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
     1321 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
     1381 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     1441 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     1501 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     1561 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     1621 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     1681 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     1741 attgattact attaataact aatgcatggc ggtaatacgg ttatccacag aatcagggga
     1801 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1861 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1921 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1981 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2041 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2101 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2161 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2221 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2281 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2341 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2401 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2461 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2521 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2581 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc
     2641 tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg
     2701 ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga
     2761 gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt
     2821 attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt
     2881 ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     2941 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     3001 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
     3061 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     3121 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     3181 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     3241 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     3301 ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt
     3361 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     3421 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     3481 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     3541 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     3601 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     3661 cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg
     3721 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     3781 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     3841 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc
     3901 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     3961 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     4021 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     4081 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     4141 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     4201 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagga
     4261 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4321 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4381 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4441 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4501 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4561 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4621 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4681 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4741 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     4801 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttacgc cttaagatac
     4861 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     4921 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     4981 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     5041 aagtaaaacc tctacaaatg tggtatggct gattatgatc tagccgctag atcgagatat
     5101 ccatgtctag atccggaccg tcccccaggc tgacatcggt cgggggggcc tttctcttct
     5161 ttttatccat ggtgggggca accttcctct tcttcttagg aggggtaccc agtcgacgcc
     5221 ccctgcggcc gctcttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     5281 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg gcggactggg
     5341 tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     5401 ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaa
//
