LOCUS       sequence-320306-        8692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Punc-4::rtetR-QFAD(650-777+790-816)::P2A::mKate::T2A::tetR-pie1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8692)
  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 8692)
  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: TC562
FEATURES             Location/Qualifiers
     source          1..8692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      71..659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     560..579
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4503..4559
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5418..5471
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(7322..7344)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7444..7463
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7541..7559)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7627..7731
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7732..7800,7801..8592)
                     /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(7950..7969)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
       61 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      121 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      181 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
      241 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      301 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      361 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      421 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc
      481 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      541 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      601 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      661 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      721 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgcggcc gccaacaata
      781 taggatgctc aggagactat aattcaaaaa acccaaccca aattagcata agcttcccca
      841 aattggaaca gtgaaatact caatcaatca ctctggcggt tttctccaca aagaacacga
      901 aattggaaaa aggaaaagat tgatttgttt gaaggcgtga ggaaatcgaa aaaaaattgt
      961 ttcaaaacaa atgaaatatt ttaacgaata aacccaggaa tgccactttt tggtgcccgc
     1021 ctacaaaaaa gtcaactgtt cttaaactcc tacatagttg ccctctctaa cgaatgttgc
     1081 tttttttaaa ttttcattga atttgattat cgtctgtatg cgcctactct atcagtggaa
     1141 aacatgccca tttcccacaa atttactcgt ttggagctca aaatgttaaa tggggcatca
     1201 gggaattggc atctcacaga aagtggcgga tgagttatat gtaataataa tggtggctga
     1261 agtggaaacc gcaaaaaaag gatgtgtatt tgagattttg caacggaatt tgaatcagat
     1321 acacaaaatg agagaaattt atagattctt taaaatccac ctgaagtcgg caaatttttg
     1381 aaaggtctgt tcgttcagtt tttcataagt tttatgaatt ttgatttact actgaagttc
     1441 gagaaaagcc ctttaaccat accaataaga aattatgtta actttacaag aatattaaca
     1501 aaaaattatt aaaaaactgt gttcccctgg cattatgata tttccaccgt cagttctcta
     1561 ttaaaacttc aactctacac agtttcaact ctcaaatgag ctgtccatct tttgccttca
     1621 ggaaattatg ctaatctgct gatttcgcgc actttcacca cactatttcc aaatagaagg
     1681 aggggtggtg caacacacga gacactgcaa aaaggttcag tgaagattta tctacttgtg
     1741 agatgagagc aggaggggtg ggagaagtgt gatgattgac gggatgacaa gaacagtgat
     1801 gtggtgtcga aatggaagat gcgctaattt gtggagtttt atgaaatttg aataattaca
     1861 tttataacat ctactagtta ggcgtctgtg taggtattgc ctgcctgcct acaaggcata
     1921 tgtacgaggc aggttaaacc tcctagaaag ctacggtact ttcaaaccac ataacattta
     1981 tactggtgtc ttttcgcgtc ttcttcttga acatgtagac cctcaaaaag ctaagtttta
     2041 cactgtgtag tttgctttga aatctacaaa aattggaggc acgccttctg gactttgccg
     2101 acacgcctac tttctaccta ccaactgcct acttccacag tctagcgtga actccaatac
     2161 cattttaaaa attgttttat ctcaaatttt caatttttaa atccaatttc ccagttctaa
     2221 aaaaaaaatt ttttttttaa tttaaaaact ctcatccttt ttaatgctcg ttcccattcg
     2281 gcaagacgtg taccaaacac aaaacaaatt gactggtgat ccatctcgaa tttctcgttt
     2341 caaattagtc gttgctatat atattgtctt atgtgtggaa aacgaataag aaaaattaga
     2401 gaaagatgag agagcagggg cagaatgaaa atttgcatgg aaattgaaga agatcactgc
     2461 tttttccgat gaatgatgac taaatagaat gagttgagag gaaaataatc ataattctaa
     2521 ttttaattag aaatatgata tggaaagtaa gcaaaaccac aaggcttttg ctctgaactt
     2581 atttccgaaa aaaaaagtga aactgaaagt tagaattttc aaaataaata ttgcgagagt
     2641 caaccttcag aaaaattggc gagaaatcaa atttgaattc tctgaaaatt tagttaaaat
     2701 acatatgtta tccttctttg ttttttggcc ttagcctgcc ttcaaggcga cctgagccta
     2761 tgcgggtgcc taataccgaa acacacataa tttttttttt taatctttaa atactgtaac
     2821 gagtcgcgag atccagcgag acccatagaa tatgttgagc gctgcaaaca atcgtataac
     2881 tcttgtccaa atttttccac tctgtatttt tgcaagcagt tcaacaaatg gtcatcattt
     2941 ttataaccca tatctatatt cgaatctgac ataacaaata atactaagca catcctccca
     3001 catctctttt cattcaagtt gcgcaattcc cctaataact atggatggag gggggaccga
     3061 gcccaaagat tattcaaaac gataacgact ttttcatgcg gccaacgaaa aataaaactt
     3121 ttctcatatt ctcctatttc ccatttcaag aattacatcc ccatgggacg ttgaaatcac
     3181 atcatattag tatatgttaa ttgttctttt cactggaata tcgccacgcc ccccttccgg
     3241 tggctcattg gtttcgagaa gaaggcggag ccgaagtggg ttcttctttc ttttttttgt
     3301 ggtagaagtc tagcaggatt aagctactat tccattagag gatcttcttc ttccaaaaag
     3361 tgaaaatgtc acgactggat aaatcgaagg tcatcaactc tgctctggaa ctgctgaatg
     3421 gagtgggaat tgaaggactt acaacccgaa agcttgcaca aaaattggga gttgaacaac
     3481 caacacttta ctggcatgtt aagaataagc gagcactttt ggatgctctt ccaattgaaa
     3541 tgttggatag acatcatact cattcatgtc cacttgaagg agaatcttgg caagatttct
     3601 tgagaaacaa cgcaaagtca taccgatgcg ctcttttgtc tcatagagat ggagcaaaag
     3661 ttcatcttgg aacacgacca actgaaaagc aatacgaaac attggaaaac caacttgctt
     3721 tcttgtgtca acaaggattt tcattggaaa atgctcttta cgcattgtct gctgttggac
     3781 attttacact tggatgcgtt ttggaagaac aagaacatca agttgcaaaa gaagaacgag
     3841 aaactccaac aactgattca atgccaccac ttttgaagca agctatcgaa cttttcgata
     3901 gacaaggagc agaaccagct ttcctttttg gactggaact tattatctgt ggacttgaga
     3961 aacaactgaa atgtgaatcg ggaggaggat ctggatctgg atctggatct ggttccggtc
     4021 gacaacttga gcttccgccg actgcgactg ccactgcaag tatcatgccc catgtcatgg
     4081 aacagcccct gtcaacgtca ataaatccgg tcaatgatcg gtttaatgga ataccgaatc
     4141 ctacccccta caatagcgat gcggccctag acgcaattac tcaaaccaat gactacggat
     4201 cagtcaatac ccatggcatt ctcagtacat atccaccacc tgccacccac ttgaacgagg
     4261 cctcggtggc ccttgcgcct ggaggtgctc ctcccagacc acccccgcca tacgttgata
     4321 gtacaacgaa ccatcctcct taccatagta acctcgtccc catggccaat ttcggatatt
     4381 cgactgtaga ctatgatgcg atggtggttg acgttgatcc gcagttcatg acgaacctcg
     4441 ggtttgtccc agggtgtaat tttagcgata tcaacacgta tgagcaacca tggggcagtg
     4501 gggctactaa cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggaccta
     4561 tggtctccga gctcattaaa gaaaacatgc atatgaagct ctacatggag ggaaccgtca
     4621 acaaccacca cttcaagtgc acctccgagg gagagggaaa gccatacgag ggaacccaaa
     4681 ccatgcgtat caaggccgtc gagggaggac cactcccatt cgccttcgac atcctcgcca
     4741 cctccttcat gtacggatcc aaggtaagtt taaacatata tatactaact aaccctgatt
     4801 atttaaattt tcagaccttc atcaaccaca cccaaggaat cccagacttc ttcaagcaat
     4861 ccttcccaga gggattcacc tgggagcgtg tcaccaccta cgaggacgga ggagtcctca
     4921 ccgccaccca agacacctcc ctccaagacg gatgcctcat ctacaacgtc aaggtaagtt
     4981 taaacagttc ggtactaact aaccatacat atttaaattt tcagatccgt ggagtcaact
     5041 tcccatccaa cggaccagtc atgcaaaaga agaccctcgg atgggaggcc tccaccgaga
     5101 ccctctaccc agccgacgga ggacttgagg gacgtgccga catggccctc aagctcgtcg
     5161 gaggaggaca cctcatctgc aacctcaagg taagtttaaa catgatttta ctaactaact
     5221 aatctgattt aaattttcag accacctacc gttccaagaa gccagccaag aacctcaaga
     5281 tgccaggagt ctactacgtc gaccgtcgtc ttgagcgtat caaggaggcc gacaaggaga
     5341 cctacgtcga gcaacacgag gtcgccgtcg cccgttactg cgacctccca tccaagctcg
     5401 gacaccgtgg atcgggagag ggacgaggaa gtcttttgac ctgcggagat gttgaggaga
     5461 acccaggacc aatgtcgcgt cttgataaat cgaaagttat taattcagct cttgaacttt
     5521 tgaacgaagt tggaattgaa ggattgacaa ctagaaagct tgcacaaaag ttgggagttg
     5581 aacaaccaac actttattgg catgttcgaa ataagcaaac tcttatgaac atgttgtctg
     5641 aagctattct tgcaaaacat catacaagat cagctccatt gccaactgaa tcttggcaac
     5701 aattccttca agaaaatgct ttgtcatttc gaaaagcact tttggttcat agagatggag
     5761 cacgattgca tattggaaca tctccaactc caccacaatt cgaacaagct gaagcacaac
     5821 ttagatgttt gtgcgatgct ggattctcag ttgaagaagc acttttcatc ttgcaatcaa
     5881 tctctcattt cacacttgga gctgttttgg aagaacaagc aactaaccaa atcgaaaaca
     5941 accatgttat tgatgctgca ccaccacttt tgcaagaagc tttcaatatt caagcacgaa
     6001 catctgctga aatggcattc catttcggac ttaagtcatt gatcttcgga ttttctgctc
     6061 aacttgatga aaagaaacat actccaattg aagatggaaa caagccaaag aagaaaagaa
     6121 aactgggaag tggaagtaat ggaagtagtg gatccagatc tgcgccacct tcttccgatt
     6181 gtccgccgcc gattgcctac taccatcatc atccacaaca tcagcaacaa ttcctgccat
     6241 ttccaatgcc atatttcttg gctccaccgc cgcaagctca acaaggagct ccttttccag
     6301 tgcaatatat tccacagcaa catgatttga tgaatagcca gccaatgtat gcaccaatgg
     6361 caccgacata ctactatcaa ccaattaatt cgaatggcat gcccatgaat tccaactgag
     6421 cgccggtcgc taccattacc aacttgtctg gtgtcaaaaa taataggggc cgctgtcatc
     6481 agagtaagtt taaactgagt tctactaact aacgagtaat atttaaattt tcagcatctc
     6541 gcgcccgtgc ctctgacttc taagtccaat tactcttcaa catccctaca tgctctttct
     6601 ccctgtgctc ccacccccta tttttgttat tatcaaaaaa cttctcttaa tttctttgtt
     6661 ttttagcttc ttttaagtca cctctaacaa tgaaattgtg tagattcaaa aatagaatta
     6721 attcgtaata aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc
     6781 ccaaaatcta cacaatgttc tgtgtacact tcttatgttt tttacttctg ataaattttt
     6841 ttgaaacatc atagaaaaaa ccgcacacaa aataccttat catatgttac gtttcagttt
     6901 atgaccgcaa tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca
     6961 tcgtgaaaaa gttttggagt atttttggaa tttttcaatc aagtgaaagt ttatgaaatt
     7021 aattttcctg cttttgcttt ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg
     7081 cgtttttctt gctaaaatca caagtattga tgagcacgat gcaagaaaga tcggaagaag
     7141 gtttgggttt gaggctcagt ggaaggtgag tagaagttga taatttgaaa gtggagtagt
     7201 gtctatgggg tttttgcctt aaatgacaga atacattccc aatataccaa acataactgt
     7261 ttcctactag tcggccgtac gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     7321 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     7381 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     7441 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     7501 gatgcgtaag gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat acgcctattt
     7561 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     7621 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     7681 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     7741 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     7801 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     7861 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     7921 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     7981 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     8041 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     8101 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     8161 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     8221 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     8281 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     8341 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     8401 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     8461 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     8521 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     8581 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     8641 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat ga
//
