LOCUS       sequence-329940-        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      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4433..4489
                     /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             5348..5401
                     /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(7252..7274)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7374..7393
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7471..7489)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7557..7661
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7662..7730,7731..8522)
                     /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(7880..7899)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgtacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgcggcc gccaacaata taggatgctc
      721 aggagactat aattcaaaaa acccaaccca aattagcata agcttcccca aattggaaca
      781 gtgaaatact caatcaatca ctctggcggt tttctccaca aagaacacga aattggaaaa
      841 aggaaaagat tgatttgttt gaaggcgtga ggaaatcgaa aaaaaattgt ttcaaaacaa
      901 atgaaatatt ttaacgaata aacccaggaa tgccactttt tggtgcccgc ctacaaaaaa
      961 gtcaactgtt cttaaactcc tacatagttg ccctctctaa cgaatgttgc tttttttaaa
     1021 ttttcattga atttgattat cgtctgtatg cgcctactct atcagtggaa aacatgccca
     1081 tttcccacaa atttactcgt ttggagctca aaatgttaaa tggggcatca gggaattggc
     1141 atctcacaga aagtggcgga tgagttatat gtaataataa tggtggctga agtggaaacc
     1201 gcaaaaaaag gatgtgtatt tgagattttg caacggaatt tgaatcagat acacaaaatg
     1261 agagaaattt atagattctt taaaatccac ctgaagtcgg caaatttttg aaaggtctgt
     1321 tcgttcagtt tttcataagt tttatgaatt ttgatttact actgaagttc gagaaaagcc
     1381 ctttaaccat accaataaga aattatgtta actttacaag aatattaaca aaaaattatt
     1441 aaaaaactgt gttcccctgg cattatgata tttccaccgt cagttctcta ttaaaacttc
     1501 aactctacac agtttcaact ctcaaatgag ctgtccatct tttgccttca ggaaattatg
     1561 ctaatctgct gatttcgcgc actttcacca cactatttcc aaatagaagg aggggtggtg
     1621 caacacacga gacactgcaa aaaggttcag tgaagattta tctacttgtg agatgagagc
     1681 aggaggggtg ggagaagtgt gatgattgac gggatgacaa gaacagtgat gtggtgtcga
     1741 aatggaagat gcgctaattt gtggagtttt atgaaatttg aataattaca tttataacat
     1801 ctactagtta ggcgtctgtg taggtattgc ctgcctgcct acaaggcata tgtacgaggc
     1861 aggttaaacc tcctagaaag ctacggtact ttcaaaccac ataacattta tactggtgtc
     1921 ttttcgcgtc ttcttcttga acatgtagac cctcaaaaag ctaagtttta cactgtgtag
     1981 tttgctttga aatctacaaa aattggaggc acgccttctg gactttgccg acacgcctac
     2041 tttctaccta ccaactgcct acttccacag tctagcgtga actccaatac cattttaaaa
     2101 attgttttat ctcaaatttt caatttttaa atccaatttc ccagttctaa aaaaaaaatt
     2161 tttttttaat ttaaaaactc tcatcctttt taatgctcgt tcccattcgg caagacgtgt
     2221 accaaacaca aaacaaattg actggtgatc catctcgaat ttctcgtttc aaattagtcg
     2281 ttgctatata tattgtctta tgtgtggaaa acgaataaga aaaattagag aaagatgaga
     2341 gagcaggggc agaatgaaaa tttgcatgga aattgaagaa gatcactgct ttttccgatg
     2401 aatgatgact aaatagaatg agttgagagg aaaataatca taattctaat tttaattaga
     2461 aatatgatat ggaaagtaag caaaaccaca aggcttttgc tctgaactta tttccgaaaa
     2521 aaaaaagtga aactgaaagt tagaattttc aaaataaata ttgcgagagt caaccttcag
     2581 aaaaattggc gagaaatcaa atttgaattc tctgaaaatt tagttaaaat acatatgtta
     2641 tccttctttg ttttttggcc ttagcctgcc ttcaaggcga cctgagccta tgcgggtgcc
     2701 taataccgaa acacacataa tttttttttt taatctttaa atactgtaac gagtcgcgag
     2761 atccagcgag acccatagaa tatgttgagc gctgcaaaca atcgtataac tcttgtccaa
     2821 atttttccac tctgtatttt tgcaagcagt tcaacaaatg gtcatcattt ttataaccca
     2881 tatctatatt cgaatctgac ataacaaata atactaagca catcctccca catctctttt
     2941 cattcaagtt gcgcaattcc cctaataact atggatggag gggggaccga gcccaaagat
     3001 tattcaaaac gataacgact ttttcatgcg gccaacgaaa aataaaactt ttctcatatt
     3061 ctcctatttc ccatttcaag aattacatcc ccatgggacg ttgaaatcac atcatattag
     3121 tatatgttaa ttgttctttt cactggaata tcgccacgcc ccccttccgg tggctcattg
     3181 gtttcgagaa gaaggcggag ccgaagtggg ttcttctttc ttttttttgt ggtagaagtc
     3241 tagcaggatt aagctactat tccattagag gatcttcttc ttccaaaaag tgaaaatgtc
     3301 acgactggat aaatcgaagg tcatcaactc tgctctggaa ctgctgaatg gagtgggaat
     3361 tgaaggactt acaacccgaa agcttgcaca aaaattggga gttgaacaac caacacttta
     3421 ctggcatgtt aagaataagc gagcactttt ggatgctctt ccaattgaaa tgttggatag
     3481 acatcatact cattcatgtc cacttgaagg agaatcttgg caagatttct tgagaaacaa
     3541 cgcaaagtca taccgatgcg ctcttttgtc tcatagagat ggagcaaaag ttcatcttgg
     3601 aacacgacca actgaaaagc aatacgaaac attggaaaac caacttgctt tcttgtgtca
     3661 acaaggattt tcattggaaa atgctcttta cgcattgtct gctgttggac attttacact
     3721 tggatgcgtt ttggaagaac aagaacatca agttgcaaaa gaagaacgag aaactccaac
     3781 aactgattca atgccaccac ttttgaagca agctatcgaa cttttcgata gacaaggagc
     3841 agaaccagct ttcctttttg gactggaact tattatctgt ggacttgaga aacaactgaa
     3901 atgtgaatcg ggaggaggat ctggatctgg atctggatct ggttccggtc gacaacttga
     3961 gcttccgccg actgcgactg ccactgcaag tatcatgccc catgtcatgg aacagcccct
     4021 gtcaacgtca ataaatccgg tcaatgatcg gtttaatgga ataccgaatc ctacccccta
     4081 caatagcgat gcggccctag acgcaattac tcaaaccaat gactacggat cagtcaatac
     4141 ccatggcatt ctcagtacat atccaccacc tgccacccac ttgaacgagg cctcggtggc
     4201 ccttgcgcct ggaggtgctc ctcccagacc acccccgcca tacgttgata gtacaacgaa
     4261 ccatcctcct taccatagta acctcgtccc catggccaat ttcggatatt cgactgtaga
     4321 ctatgatgcg atggtggttg acgttgatcc gcagttcatg acgaacctcg ggtttgtccc
     4381 agggtgtaat tttagcgata tcaacacgta tgagcaacca tggggcagtg gggctactaa
     4441 cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggaccta tggtctccga
     4501 gctcattaaa gaaaacatgc atatgaagct ctacatggag ggaaccgtca acaaccacca
     4561 cttcaagtgc acctccgagg gagagggaaa gccatacgag ggaacccaaa ccatgcgtat
     4621 caaggccgtc gagggaggac cactcccatt cgccttcgac atcctcgcca cctccttcat
     4681 gtacggatcc aaggtaagtt taaacatata tatactaact aaccctgatt atttaaattt
     4741 tcagaccttc atcaaccaca cccaaggaat cccagacttc ttcaagcaat ccttcccaga
     4801 gggattcacc tgggagcgtg tcaccaccta cgaggacgga ggagtcctca ccgccaccca
     4861 agacacctcc ctccaagacg gatgcctcat ctacaacgtc aaggtaagtt taaacagttc
     4921 ggtactaact aaccatacat atttaaattt tcagatccgt ggagtcaact tcccatccaa
     4981 cggaccagtc atgcaaaaga agaccctcgg atgggaggcc tccaccgaga ccctctaccc
     5041 agccgacgga ggacttgagg gacgtgccga catggccctc aagctcgtcg gaggaggaca
     5101 cctcatctgc aacctcaagg taagtttaaa catgatttta ctaactaact aatctgattt
     5161 aaattttcag accacctacc gttccaagaa gccagccaag aacctcaaga tgccaggagt
     5221 ctactacgtc gaccgtcgtc ttgagcgtat caaggaggcc gacaaggaga cctacgtcga
     5281 gcaacacgag gtcgccgtcg cccgttactg cgacctccca tccaagctcg gacaccgtgg
     5341 atcgggagag ggacgaggaa gtcttttgac ctgcggagat gttgaggaga acccaggacc
     5401 aatgtcgcgt cttgataaat cgaaagttat taattcagct cttgaacttt tgaacgaagt
     5461 tggaattgaa ggattgacaa ctagaaagct tgcacaaaag ttgggagttg aacaaccaac
     5521 actttattgg catgttcgaa ataagcaaac tcttatgaac atgttgtctg aagctattct
     5581 tgcaaaacat catacaagat cagctccatt gccaactgaa tcttggcaac aattccttca
     5641 agaaaatgct ttgtcatttc gaaaagcact tttggttcat agagatggag cacgattgca
     5701 tattggaaca tctccaactc caccacaatt cgaacaagct gaagcacaac ttagatgttt
     5761 gtgcgatgct ggattctcag ttgaagaagc acttttcatc ttgcaatcaa tctctcattt
     5821 cacacttgga gctgttttgg aagaacaagc aactaaccaa atcgaaaaca accatgttat
     5881 tgatgctgca ccaccacttt tgcaagaagc tttcaatatt caagcacgaa catctgctga
     5941 aatggcattc catttcggac ttaagtcatt gatcttcgga ttttctgctc aacttgatga
     6001 aaagaaacat actccaattg aagatggaaa caagccaaag aagaaaagaa aactgggaag
     6061 tggaagtaat ggaagtagtg gatccagatc tgcgccacct tcttccgatt gtccgccgcc
     6121 gattgcctac taccatcatc atccacaaca tcagcaacaa ttcctgccat ttccaatgcc
     6181 atatttcttg gctccaccgc cgcaagctca acaaggagct ccttttccag tgcaatatat
     6241 tccacagcaa catgatttga tgaatagcca gccaatgtat gcaccaatgg caccgacata
     6301 ctactatcaa ccaattaatt cgaatggcat gcccatgaat tccaactgag cgccggtcgc
     6361 taccattacc aacttgtctg gtgtcaaaaa taataggggc cgctgtcatc agagtaagtt
     6421 taaactgagt tctactaact aacgagtaat atttaaattt tcagcatctc gcgcccgtgc
     6481 ctctgacttc taagtccaat tactcttcaa catccctaca tgctctttct ccctgtgctc
     6541 ccacccccta tttttgttat tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc
     6601 ttttaagtca cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata
     6661 aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta
     6721 cacaatgttc tgtgtacact tcttatgttt tttacttctg ataaattttt ttgaaacatc
     6781 atagaaaaaa ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa
     6841 tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa
     6901 gttttggagt atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg
     6961 cttttgcttt ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt
     7021 gctaaaatca caagtattga tgagcacgat gcaagaaaga tcggaagaag gtttgggttt
     7081 gaggctcagt ggaaggtgag tagaagttga taatttgaaa gtggagtagt gtctatgggg
     7141 tttttgcctt aaatgacaga atacattccc aatataccaa acataactgt ttcctactag
     7201 tcggccgtac gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     7261 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     7321 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     7381 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     7441 gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat acgcctattt ttataggtta
     7501 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     7561 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     7621 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     7681 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     7741 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     7801 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     7861 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     7921 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     7981 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     8041 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     8101 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     8161 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     8221 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     8281 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     8341 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     8401 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     8461 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     8521 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     8581 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     8641 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
