LOCUS       sequence-329339-        6186 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 6186)
  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 6186)
  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..6186
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    95..116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(131..136,137..154,155..161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    169..185
                     /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     174..196
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     193..209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     193..209
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1301..1321
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2030..2041
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2334..3044
                     /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(2479..2497)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2747..2766
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3960..3982)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4082..4101
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4183..4201)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4269..4373
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4374..4442,4443..5234)
                     /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(4592..4611)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5405..5993
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5894..5913
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6147..6164
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      181 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca tgcgcggccg
      241 ctgactggtg atccatctcg aatttctcgt ttcaaattag tcgttgctat atatattgtc
      301 ttatgtgtgg aaaacgaata agaaaaatta gagaaagatg agagagcagg ggcagaatga
      361 aaatttgcat ggaaattgaa gaagatcact gctttttccg atgaatgatg actaaataga
      421 atgagttgag aggaaaataa tcataattct aattttaatt agaaatatga tatggaaagt
      481 aagcaaaacc acaaggcttt tgctctgaac ttatttccga aaaaaaaagt gaaactgaaa
      541 gttagaattt tcaaaataaa tattgcgaga gtcaaccttc agaaaaattg gcgagaaatc
      601 aaatttgaat tctctgaaaa tttagttaaa atacatatgt tatccttctt tgttttttgg
      661 ccttagcctg ccttcaaggc gacctgagcc tatgcgggtg cctaataccg aaacacacat
      721 aatttttttt tttaatcttt aaatactgta acgagtcgcg agatccagcg agacccatag
      781 aatatgttga gcgctgcaaa caatcgtata actcttgtcc aaatttttcc actctgtatt
      841 tttgcaagca gttcaacaaa tggtcatcat ttttataacc catatctata ttcgaatctg
      901 acataacaaa taatactaag cacatcctcc cacatctctt ttcattcaag ttgcgcaatt
      961 cccctaataa ctatggatgg aggggggacc gagcccaaag attattcaaa acgataacga
     1021 ctttttcatg cggccaacga aaaataaaac ttttctcata ttctcctatt tcccatttca
     1081 agaattacat ccccatggga cgttgaaatc acatcatatt agtatatgtt aattgttctt
     1141 ttcactggaa tatcgccacg ccccccttcc ggtggctcat tggtttcgag aagaaggcgg
     1201 agccgaagtg ggttcttctt tctttttttt gtggtagaag tctagcagga ttaagctact
     1261 attccattag aggatcttct tcttccaaaa agtgaaaatg ccaaagaaga agcgtaaggt
     1321 catgtcacga ctggataaat cgaaggtcat caactctgct ctggaactgc tgaatggagt
     1381 gggaattgaa ggacttacaa cccgaaagct tgcacaaaaa ttgggagttg aacaaccaac
     1441 actttactgg catgttaaga ataagcgagc acttttggat gctcttccaa ttgaaatgtt
     1501 ggatagacat catactcatt catgtccact tgaaggagaa tcttggcaag atttcttgag
     1561 aaacaacgca aagtcatacc gatgcgctct tttgtctcat agagatggag caaaagttca
     1621 tcttggaaca cgaccaactg aaaagcaata cgaaacattg gaaaaccaac ttgctttctt
     1681 gtgtcaacaa ggattttcat tggaaaatgc tctttacgca ttgtctgctg ttggacattt
     1741 tacacttgga tgcgttttgg aagaacaaga acatcaagtt gcaaaagaag aacgagaaac
     1801 tccaacaact gattcaatgc caccactttt gaagcaagct atcgaacttt tcgatagaca
     1861 aggagcagaa ccagctttcc tttttggact ggaacttatt atctgtggac ttgagaaaca
     1921 actgaaatgt gaatcgggag gaggatccgg atcaggatct ggatcaggat ctggatcagc
     1981 gcttaaaaaa gaattgcaag ctaataagaa ggaacttgca caattgaagt gggaacttca
     2041 agctttgaaa aaagaacttg cacagtaagc tgtctcatcc tactttcacc tagttaactg
     2101 cttgtcttaa aatctatgct tctctttagt atctaaaatt ttcctagaag cttacaagta
     2161 tataaatggt ctcttctcaa taaaggttgt atatttattc atcttattga atctgccatt
     2221 tcctcgtttt tgcgagttta tataccttcc aattttcttt ctattgtatt ttcaacttct
     2281 aattttaatt cagggaaact gcttcaacgc atccggtaga aaaaatgctc gagatggtga
     2341 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     2401 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     2461 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     2521 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     2581 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     2641 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     2701 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     2761 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     2821 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     2881 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     2941 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     3001 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagaattc caactgagcg
     3061 ccggtcgcta ccattaccaa cttgtctggt gtcaaaaata ataggggccg ctgtcatcag
     3121 agtaagttta aactgagttc tactaactaa cgagtaatat ttaaattttc agcatctcgc
     3181 gcccgtgcct ctgacttcta agtccaatta ctcttcaaca tccctacatg ctctttctcc
     3241 ctgtgctccc accccctatt tttgttatta tcaaaaaact tctcttaatt tctttgtttt
     3301 ttagcttctt ttaagtcacc tctaacaatg aaattgtgta gattcaaaaa tagaattaat
     3361 tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc attaataata attctatccc
     3421 aaaatctaca caatgttctg tgtacacttc ttatgttttt tacttctgat aaattttttt
     3481 gaaacatcat agaaaaaacc gcacacaaaa taccttatca tatgttacgt ttcagtttat
     3541 gaccgcaatt tttatttctt cgcacgtctg ggcctctcat gacgtcaaat catgctcatc
     3601 gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa gtgaaagttt atgaaattaa
     3661 ttttcctgct tttgcttttt ggggtttccc ctattgtttg tcaagatttc gaggacggcg
     3721 tttttcttgc taaaatcaca agtattgatg agcacgatgc aagaaagatc ggaagaaggt
     3781 ttgggtttga ggctcagtgg aaggtgagta gaagttgata atttgaaagt ggagtagtgt
     3841 ctatggggtt tttgccttaa atgacagaat acattcccaa tataccaaac ataactgttt
     3901 cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     3961 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4021 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     4081 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     4141 tgcgtaagga gaaaataccg catcaggcgg ccttaattaa gggcctcgtg atacgcctat
     4201 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4261 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4321 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4381 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4441 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4501 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4561 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4621 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4681 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4741 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4801 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4861 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4921 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4981 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5041 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5101 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5161 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5221 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5281 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5341 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5401 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5461 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5521 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5581 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5641 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5701 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5761 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5821 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5881 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5941 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6001 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6061 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6121 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     6181 caatac
//
