LOCUS       sequence-244028-        6186 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Punc-4c::SV40 NLS::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 Apr 14, 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    85..106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(121..126,127..144,145..151)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    159..175
                     /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     164..186
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     183..199
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     183..199
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1291..1311
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2020..2031
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2324..3034
                     /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(2469..2487)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2737..2756
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3950..3972)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4072..4091
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4173..4191)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4259..4363
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4364..4432,4433..5224)
                     /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(4582..4601)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5395..5983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5884..5903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6137..6154
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
       61 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      121 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      181 cacaggaaac agctatgacc atgattacgc caagcttgca tgcgcggccg ctgactggtg
      241 atccatctcg aatttctcgt ttcaaattag tcgttgctat atatattgtc ttatgtgtgg
      301 aaaacgaata agaaaaatta gagaaagatg agagagcagg ggcagaatga aaatttgcat
      361 ggaaattgaa gaagatcact gctttttccg atgaatgatg actaaataga atgagttgag
      421 aggaaaataa tcataattct aattttaatt agaaatatga tatggaaagt aagcaaaacc
      481 acaaggcttt tgctctgaac ttatttccga aaaaaaaagt gaaactgaaa gttagaattt
      541 tcaaaataaa tattgcgaga gtcaaccttc agaaaaattg gcgagaaatc aaatttgaat
      601 tctctgaaaa tttagttaaa atacatatgt tatccttctt tgttttttgg ccttagcctg
      661 ccttcaaggc gacctgagcc tatgcgggtg cctaataccg aaacacacat aatttttttt
      721 tttaatcttt aaatactgta acgagtcgcg agatccagcg agacccatag aatatgttga
      781 gcgctgcaaa caatcgtata actcttgtcc aaatttttcc actctgtatt tttgcaagca
      841 gttcaacaaa tggtcatcat ttttataacc catatctata ttcgaatctg acataacaaa
      901 taatactaag cacatcctcc cacatctctt ttcattcaag ttgcgcaatt cccctaataa
      961 ctatggatgg aggggggacc gagcccaaag attattcaaa acgataacga ctttttcatg
     1021 cggccaacga aaaataaaac ttttctcata ttctcctatt tcccatttca agaattacat
     1081 ccccatggga cgttgaaatc acatcatatt agtatatgtt aattgttctt ttcactggaa
     1141 tatcgccacg ccccccttcc ggtggctcat tggtttcgag aagaaggcgg agccgaagtg
     1201 ggttcttctt tctttttttt gtggtagaag tctagcagga ttaagctact attccattag
     1261 aggatcttct tcttccaaaa agtgaaaatg ccaaagaaga agcgtaaggt catgtcacga
     1321 ctggataaat cgaaggtcat caactctgct ctggaactgc tgaatggagt gggaattgaa
     1381 ggacttacaa cccgaaagct tgcacaaaaa ttgggagttg aacaaccaac actttactgg
     1441 catgttaaga ataagcgagc acttttggat gctcttccaa ttgaaatgtt ggatagacat
     1501 catactcatt catgtccact tgaaggagaa tcttggcaag atttcttgag aaacaacgca
     1561 aagtcatacc gatgcgctct tttgtctcat agagatggag caaaagttca tcttggaaca
     1621 cgaccaactg aaaagcaata cgaaacattg gaaaaccaac ttgctttctt gtgtcaacaa
     1681 ggattttcat tggaaaatgc tctttacgca ttgtctgctg ttggacattt tacacttgga
     1741 tgcgttttgg aagaacaaga acatcaagtt gcaaaagaag aacgagaaac tccaacaact
     1801 gattcaatgc caccactttt gaagcaagct atcgaacttt tcgatagaca aggagcagaa
     1861 ccagctttcc tttttggact ggaacttatt atctgtggac ttgagaaaca actgaaatgt
     1921 gaatcgggag gaggatccgg atcaggatct ggatcaggat ctggatcagc gcttaaaaaa
     1981 gaattgcaag ctaataagaa ggaacttgca caattgaagt gggaacttca agctttgaaa
     2041 aaagaacttg cacagtaagc tgtctcatcc tactttcacc tagttaactg cttgtcttaa
     2101 aatctatgct tctctttagt atctaaaatt ttcctagaag cttacaagta tataaatggt
     2161 ctcttctcaa taaaggttgt atatttattc atcttattga atctgccatt tcctcgtttt
     2221 tgcgagttta tataccttcc aattttcttt ctattgtatt ttcaacttct aattttaatt
     2281 cagggaaact gcttcaacgc atccggtaga aaaaatgctc gagatggtga gcaagggcga
     2341 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     2401 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     2461 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     2521 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     2581 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     2641 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     2701 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     2761 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     2821 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     2881 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     2941 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     3001 ctccaccggc ggcatggacg agctgtacaa gtaagaattc caactgagcg ccggtcgcta
     3061 ccattaccaa cttgtctggt gtcaaaaata ataggggccg ctgtcatcag agtaagttta
     3121 aactgagttc tactaactaa cgagtaatat ttaaattttc agcatctcgc gcccgtgcct
     3181 ctgacttcta agtccaatta ctcttcaaca tccctacatg ctctttctcc ctgtgctccc
     3241 accccctatt tttgttatta tcaaaaaact tctcttaatt tctttgtttt ttagcttctt
     3301 ttaagtcacc tctaacaatg aaattgtgta gattcaaaaa tagaattaat tcgtaataaa
     3361 aagtcgaaaa aaattgtgct ccctcccccc attaataata attctatccc aaaatctaca
     3421 caatgttctg tgtacacttc ttatgttttt tacttctgat aaattttttt gaaacatcat
     3481 agaaaaaacc gcacacaaaa taccttatca tatgttacgt ttcagtttat gaccgcaatt
     3541 tttatttctt cgcacgtctg ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt
     3601 tttggagtat ttttggaatt tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct
     3661 tttgcttttt ggggtttccc ctattgtttg tcaagatttc gaggacggcg tttttcttgc
     3721 taaaatcaca agtattgatg agcacgatgc aagaaagatc ggaagaaggt ttgggtttga
     3781 ggctcagtgg aaggtgagta gaagttgata atttgaaagt ggagtagtgt ctatggggtt
     3841 tttgccttaa atgacagaat acattcccaa tataccaaac ataactgttt cctactagtc
     3901 ggccgtacgg gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3961 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     4021 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     4081 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     4141 gaaaataccg catcaggcgg ccttaattaa gggcctcgtg atacgcctat ttttataggt
     4201 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4261 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4321 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4381 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4441 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4501 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4561 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4621 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4681 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4741 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4801 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4861 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4921 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4981 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5041 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5101 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5161 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5221 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5281 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5341 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5401 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5461 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5521 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5581 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5641 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5701 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5761 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5821 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5881 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5941 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6001 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6061 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6121 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     6181 accgcc
//
