LOCUS       sequence-320308-        6245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Punc-4c::SV40NLS::rtetR::Zip(+)::SL2::mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6245)
  AUTHORS   Mao S, Qi Y, Zhu H, Huang X, Zou Y, Chi T
  TITLE     A Tet/Q Hybrid System for Robust and Versatile Control of Transgene 
            Expression in C. elegans.
  JOURNAL   iScience. 2019 Jan 25;11:224-237. doi: 10.1016/j.isci.2018.12.023. 
            Epub 2018 Dec 27.
  PUBMED    30634168
REFERENCE   2  (bases 1 to 6245)
  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: "iScience.";
            date: "2019-01-25"; volume: "11:224-237. doi"; pages: " 
            10.1016/j.isci.2018.12.023. Epub 2018 Dec 27"
COMMENT     Sequence Label: TC513
FEATURES             Location/Qualifiers
     source          1..6245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      208..796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     697..716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     950..967
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1084..1105
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1120..1125,1126..1143,1144..1150)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1158..1174
                     /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)."
     primer_bind     1163..1185
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             2343..2363
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3072..3083
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             3367..4077
                     /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(3512..3530)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3780..3799
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(5008..5030)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5130..5149
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5231..5249)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5317..5421
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5422..5490,5491..6245,1..37)
                     /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     complement(5640..5659)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
       61 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
      121 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
      181 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      241 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
      301 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
      361 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
      421 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
      481 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
      541 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca
      601 ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
      661 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
      721 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      781 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      841 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      901 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      961 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     1021 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     1081 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     1141 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctgggggtg
     1201 gggggggggt tgggggtggg gtgttgttgd gggtgtgtdd tttgtdatga ccatgattac
     1261 gccaagcttg catgcgcggc cgctgactgg tgatccatct cgaatttctc gtttcaaatt
     1321 agtcgttgct atatatattg tcttatgtgt ggaaaacgaa taagaaaaat tagagaaaga
     1381 tgagagagca ggggcagaat gaaaatttgc atggaaattg aagaagatca ctgctttttc
     1441 cgatgaatga tgactaaata gaatgagttg agaggaaaat aatcataatt ctaattttaa
     1501 ttagaaatat gatatggaaa gtaagcaaaa ccacaaggct tttgctctga acttatttcc
     1561 gaaaaaaaaa gtgaaactga aagttagaat tttcaaaata aatattgcga gagtcaacct
     1621 tcagaaaaat tggcgagaaa tcaaatttga attctctgaa aatttagtta aaatacatat
     1681 gttatccttc tttgtttttt ggccttagcc tgccttcaag gcgacctgag cctatgcggg
     1741 tgcctaatac cgaaacacac ataatttttt tttttaatct ttaaatactg taacgagtcg
     1801 cgagatccag cgagacccat agaatatgtt gagcgctgca aacaatcgta taactcttgt
     1861 ccaaattttt ccactctgta tttttgcaag cagttcaaca aatggtcatc atttttataa
     1921 cccatatcta tattcgaatc tgacataaca aataatacta agcacatcct cccacatctc
     1981 ttttcattca agttgcgcaa ttcccctaat aactatggat ggagggggga ccgagcccaa
     2041 agattattca aaacgataac gactttttca tgcggccaac gaaaaataaa acttttctca
     2101 tattctccta tttcccattt caagaattac atccccatgg gacgttgaaa tcacatcata
     2161 ttagtatatg ttaattgttc ttttcactgg aatatcgcca cgcccccctt ccggtggctc
     2221 attggtttcg agaagaaggc ggagccgaag tgggttcttc tttctttttt ttgtggtaga
     2281 agtctagcag gattaagcta ctattccatt agaggatctt cttcttccaa aaagtgaaaa
     2341 tgccaaagaa gaagcgtaag gtcatgtcac gactggataa atcgaaggtc atcaactctg
     2401 ctctggaact gctgaatgga gtgggaattg aaggacttac aacccgaaag cttgcacaaa
     2461 aattgggagt tgaacaacca acactttact ggcatgttaa gaataagcga gcacttttgg
     2521 atgctcttcc aattgaaatg ttggatagac atcatactca ttcatgtcca cttgaaggag
     2581 aatcttggca agatttcttg agaaacaacg caaagtcata ccgatgcgct cttttgtctc
     2641 atagagatgg agcaaaagtt catcttggaa cacgaccaac tgaaaagcaa tacgaaacat
     2701 tggaaaacca acttgctttc ttgtgtcaac aaggattttc attggaaaat gctctttacg
     2761 cattgtctgc tgttggacat tttacacttg gatgcgtttt ggaagaacaa gaacatcaag
     2821 ttgcaaaaga agaacgagaa actccaacaa ctgattcaat gccaccactt ttgaagcaag
     2881 ctatcgaact tttcgataga caaggagcag aaccagcttt cctttttgga ctggaactta
     2941 ttatctgtgg acttgagaaa caactgaaat gtgaatcggg aggaggatcc ggatcaggat
     3001 ctggatcagg atctggatca gcgcttaaaa aagaattgca agctaataag aaggaacttg
     3061 cacaattgaa gtgggaactt caagctttga aaaaagaact tgcacagtaa gctgtctcat
     3121 cctactttca cctagttaac tgcttgtctt aaaatctatg cttctcttta gtatctaaaa
     3181 ttttcctaga agcttacaag tatataaatg gtctcttctc aataaaggtt gtatatttat
     3241 tcatcttatt gaatctgcca tttcctcgtt tttgcgagtt tatatacctt ccaattttct
     3301 ttctattgta ttttcaactt ctaattttaa ttcagggaaa ctgcttcaac gcatccggta
     3361 gaaaaaatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     3421 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     3481 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     3541 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     3601 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     3661 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     3721 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     3781 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     3841 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     3901 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     3961 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     4021 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaacat
     4081 tcgtagaatt ccaactgagc gccggtcgct accattacca acttgtctgg tgtcaaaaat
     4141 aataggggcc gctgtcatca gagtaagttt aaactgagtt ctactaacta acgagtaata
     4201 tttaaatttt cagcatctcg cgcccgtgcc tctgacttct aagtccaatt actcttcaac
     4261 atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt atcaaaaaaa
     4321 cttcttctta atttctttgt tttttagctt cttttaagtc acctctaaca atgaaattgt
     4381 gtagattcaa aaatagaatt aattcgtaat aaaaagtcga aaaaaattgt gctccctccc
     4441 cccattaata ataattctat cccaaaatct acacaatgtt ctgtgtacac ttcttatgtt
     4501 ttttttactt ctgataaatt ttttttgaaa catcatagaa aaaaccgcac acaaaatacc
     4561 ttatcatatg ttacgtttca gtttatgacc gcaattttta tttcttcgca cgtctgggcc
     4621 tctcatgacg tcaaatcatg ctcatcgtga aaaagttttg gagtattttt ggaatttttc
     4681 aatcaagtga aagtttatga aattaatttt cctgcttttg ctttttgggg gtttccccta
     4741 ttgtttgtca agagtttcga ggacggcgtt tttcttgcta aaatcacaag tattgatgag
     4801 cacgatgcaa gaaagatcgg aagaaggttt gggtttgagg ctcagtggaa ggtgagtaga
     4861 agttgataat ttgaaagtgg agtagtgtct atggggtttt tgccttaaat gacagaatac
     4921 attcccaata taccaaacat aactgtttcc tactagtcgg ccgtacgggc cctttcgtct
     4981 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     5041 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     5101 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5161 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcggcc
     5221 ttaattaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5281 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5341 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5401 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5461 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5521 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5581 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5641 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5701 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5761 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5821 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5881 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5941 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6001 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6061 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6121 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6181 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6241 cagat
//
