LOCUS       sequence-380227-        7294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-inducible (TetOn) mCherry with SV40 NLS, control 
            construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7294)
  AUTHORS   Ratnayeke N, Baris Y, Chung M, Yeeles JTP, Meyer T
  TITLE     CDT1 inhibits CMG helicase in early S phase to separate origin 
            licensing from DNA synthesis.
  JOURNAL   Mol Cell. 2023 Jan 5;83(1):26-42.e13. doi: 
            10.1016/j.molcel.2022.12.004.
  PUBMED    36608667
REFERENCE   2  (bases 1 to 7294)
  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: 
            "10.1016/j.molcel.2022.12"; journalName: "Mol Cell"; date: 
            "2023-01-5- 5"; volume: "83"; issue: "1"; pages: "26-42.e13"
COMMENT     Sequence Label: pLV-TetOn-NLS-mCherry
FEATURES             Location/Qualifiers
     source          1..7294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     protein_bind    2202..2220
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2237..2255
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2273..2291
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2309..2327
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2344..2362
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2380..2398
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2410..2448
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2445..2469
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2484..2493
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2496..2516
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2529..3239
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2674..2692)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2942..2961
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3263..3851
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3316..3336)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3734..3745)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4058..4691
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4819..4835)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4819..4835)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4832..4854)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4843..4859
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4867..4873,4874..4891,4892..4897))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4912..4933
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5050..5067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5221..5809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5301..5320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5980..6771,6772..6840))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6603..6622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6841..6945)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    6993..7127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7030..7049)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7084..7103
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatagacta gacgagttta ctccctatca gtgatagaga
     2221 acgatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc
     2281 agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg tatgtcgagt
     2341 ttatccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac
     2401 gtatgtcgag gtaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac
     2461 cgtcagatcg caccggcgaa ttcgccacca tggcacctaa gaaaaagcgg aaagtgggcg
     2521 gaagtggtat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     2581 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     2641 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     2701 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     2761 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     2821 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     2881 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     2941 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     3001 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     3061 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     3121 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     3181 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtgac
     3241 ccgggcgggg cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac
     3301 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3361 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3421 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3481 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3541 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3601 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3661 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3721 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3781 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3841 cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca
     3901 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag
     3961 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct
     4021 gtagatctta gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa
     4081 cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt ccctgattag
     4141 cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag
     4201 ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg
     4261 ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg
     4321 tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag
     4381 aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt
     4441 ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt
     4501 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     4561 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     4621 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     4681 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     4741 atgaatatca gagagtgaga ggccttgaca ttgctagcgt ttaccgtcga cctctagcta
     4801 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4861 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4921 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4981 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5041 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5101 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5161 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5221 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5281 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5341 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5401 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5461 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5521 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5581 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5641 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5701 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5761 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5821 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5881 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5941 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6001 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6061 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6121 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6181 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6241 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6301 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6361 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6421 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6481 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6541 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6601 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6661 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6721 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6781 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6841 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6901 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6961 agtgccacct gacgtcgacg gatcgggaga tcaacttgtt tattgcagct tataatggtt
     7021 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     7081 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc
     7141 aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc
     7201 tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt
     7261 gtatttgtta aatatgtact acaaacttag tagt
//
