LOCUS       sequence-245108-       10332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pser-2::rtetR-QFAD::P2A::mKate::T2A::tetR-pie1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10332)
  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 10332)
  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: TC393
FEATURES             Location/Qualifiers
     source          1..10332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     266..285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             5849..5905
                     /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             6764..6817
                     /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(8668..8690)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8790..8809
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(8887..8905)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8973..9077
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9078..9146,9147..9938)
                     /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(9296..9315)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(10109..10332,1..365)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
       61 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      121 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      181 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      241 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      301 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      361 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      421 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcggccgcac
      481 tgactgggcc ggcccatgct tgttctagtg atcaaaattg tcacttataa agtaaaagtt
      541 tagtaaatta actgctaaag aatataaagc taacacagca aaacatgtga atgatccaac
      601 gataatcacc acccacaaca tagctttgag ttgattattt gagtttttaa atcaaaatcg
      661 tctttttaaa acaggataaa cccgacgaaa ttcaaaaaat tcccagaaag tgaaaaaaaa
      721 tcaagcaagt atgtgtcttt gatttttgta gttttttttt acaaatttca aaagatctag
      781 tatcctaaat tataccgtat attccctatt agtatcgccc ccctcactcg acttatcgtt
      841 tatatcttgg ttgtctttga tcatatttta aaaattcaaa cgacaaaatc tttttcaaat
      901 ctatcgttat gctctggttt ttgaagatat caaaaatctt ctgaaaccgc tgaaatatta
      961 gccgacaaag tcgattaatg gggggccata ataataggga atagtgtttt tcccatattc
     1021 cctaattgtt ttccaaatat caattttcac agttttaatt taattgcgct tactacatag
     1081 ttttaactaa gttttaatat tcgaaactag ctgttacaaa aaggttatga aattacattt
     1141 tcagacgatt tctgaatttt taaacattca aaaggtcttg ctatattgtt tgatccaaat
     1201 aaacaaaaag cctgtacaag aaacatatct tatttcatca taaataatgt tgttctattt
     1261 tgaaataatt acataattaa gttgaaaaat tgtcagaagc cggaaaaaaa cacgaatcga
     1321 ctttcccagc aagaatatca gaaaacctgt tgagcaaaaa aaacaatctc gttgttgaaa
     1381 aaaccagata gctggaagaa attgaaaata aaaagacaac acacatatat actaaaaaca
     1441 tgaggcaaaa gaactgtcta ttgaaacacg aaacgaggag aaaaaaaaat aaaaacggcc
     1501 ttgtgattgt ccagttgagt tttgtttcac acggttttta cgatcaatgc aaagaaaaaa
     1561 aaaggattca ataatggtgg aagaaatatt cgtggaagta caactatgta ctacaactta
     1621 tgaatttgaa tttcattccc gaaaatttgt cggaaaattg tgtaaaaggg gtcatataga
     1681 taataaccga tgtgggggga gaaaggatga agaagtggtg ggaggcaaaa caaagatgaa
     1741 tccgaaaagg atgcagcaat tcagaacaga tgcatcccgg aaaatgaaaa cgaaggtcat
     1801 cgctatttct taccttttca ccaaaatctt tcttcttcac tgatccaggg tcagaaattt
     1861 gaaattttga aggacaacag tgttgccata gcaggaactg ggggctgggg ctggggggag
     1921 aaaatttatt tctcacgtcg tttttctcaa ccttcttcag tctcctagat atctcctaga
     1981 aacctttgag ttatacgagt gaagagacgg aaaagccaca gaataaggga aaaatgtaag
     2041 taagagtggt ggagagccaa ggaaatgtga caaattaata acacgacggc ctacaactca
     2101 ctttgttata ataattttaa tttgaaatct tctttcttgc aaaaagagta ctagtgaagg
     2161 acggatgaag aacgatggaa tacttgaaaa tctgctcaga tcttcttctt tcctcctcga
     2221 aattagagtt aatccaaaag ataaacagtt gtaagtggtg tgtgaatctt caagagagcc
     2281 ggagatgaca gtctcagagg tgagtcactg tttttttttg ttggattgtg ctgggcttag
     2341 ctgctagccg gatgtaagaa tattgaaaaa atgaatataa aaagtttaat gatataaacc
     2401 ggtagcgaga tgatatataa gttcagatgt gttgataaat agggtttttc gtttaaatgt
     2461 actgataatt attgaaaatt ttaaattttc tagaaaatca gttcaaaaaa ttttgatttt
     2521 gtaataacaa aaaagttcag tcaaacgttc aggcagttgc catgagttta gtaaagctag
     2581 actttttgag aacatttttt gttgaaataa ttgttttaat tttaaaatgt ttagttcaaa
     2641 taactagaac atacataagt agaacataaa atttgaataa aattgtcgtc ttccaaaatt
     2701 gtaaaattat attgcaaaaa ctgtgtaact caaactgctg atcaaaatat ttattgaagt
     2761 ttaacagagt ttaaaacaca aaattatagt gaagaaatgc acagaaaatg catccaaact
     2821 caattatttc ctagaatgaa gaattttaaa aatgatgtta taacaaaatt cttttcaatt
     2881 caaaaactag attgtattta caacgtcgga atgattaaaa cacatttaga atataaaatt
     2941 cttaaaataa attaaaattt tgtggactgg actttttgga gacttggcga attatggtta
     3001 ttccgaaacg ctgtcgactt caactgtagg cgaaatacac gcgcgcggaa ctcgtaagat
     3061 gtcggcggcc ttatattggg tggggtcccg atttttgcct ttttcccgtg agtggacgag
     3121 caaaattcgg ggcttctttg ggcgtacacg ccaagcagtt ttattatttt gagtgtatag
     3181 ctcttaaaaa actgtaaaaa tgtgtttgaa aaagatatgc tatgaatttt gatcatttta
     3241 tccactgtac tgaaactcga acataaaaac caaaattata cgaaaaaatc gtttggagac
     3301 ggcgattcta acttttggtg atgaaaaact tatcgaaatg ttacatttcg aatctttgat
     3361 atcagaaaaa aagtcactga tcatcttctg aaataaatta acaagaaagt attcctctag
     3421 tatcaaaatg tctctagtgg ctttctcaaa gacattaatc cccatatctc atagttcaga
     3481 cgaactttcc acagtaaaca atactagacg ttcagaatat gaaatttaat catgttttat
     3541 atgaaaaaag ttgctccatt caattttaat actcgacaaa aaaggaacca agttccttag
     3601 tgcaaaaatt ccaaaaatct aaaacaggtt ccactgagct ttggcaaaat gattagaaga
     3661 gcatttcaat gctcttctaa tatgtttcta ggtgttattt tttttctcaa tttttttttg
     3721 taactttctt gttttagcaa caacaaaaat tatttcattt gaaattacag aaacagaaat
     3781 tctcaaaatg ctgtttccaa aaatcaggta taatttgttc acacaattaa taatgtttct
     3841 agaaaatagt tttattttga cgcaactaag aaacataaaa acatctttta atcagtcagt
     3901 tttaatcagg ggcagatagc tcagccggta gcgccggccg ctagcaatct ggaggtcacg
     3961 agttcaagtc ccgcctcacc ccctatgctc acccatcatc tattaggaag tggagcaatc
     4021 cccaactaga ttatcggcca cagtccccgg ctaggacgtg gcttaaatta cagctcagtg
     4081 ggagcaccac caggcagtgt acctgaatcc cagatccgca gtacatagca cttgaagaac
     4141 ggatcgtcct ttaatccttt ttttttaaat ctcgtttcaa aattatttta aacccatttt
     4201 tttcaaattg ttacttgaat tcagtgtttc tgaatttatt acagccatat gtgttttgtg
     4261 atttctgatc catctcgatt cggtttccac ttctcatatc catttctcaa cccgtttttt
     4321 ttgtattcta cctttttcct accaactcat gtcaatttcg cacaacataa caaatcacaa
     4381 cgtctaaaag cgaagcaaat gaatgtaaaa gaaggttatc aatcagagtc tcctcgggct
     4441 ccttcctttt gaggtttgat gacgcgaaaa atcggaaggg ccttgcgttg tcctccggcc
     4501 agtagcaatt catttagtag acgttttttt cccctgctac attctgacga caataatatt
     4561 atttttgttg gctaaattta ctatagctaa ttattcattt tggcatattt ttgtgacatc
     4621 acaacacata atgggcgcgc ctctagagga tcgctagcac gctctcgtgg atcaaaaaat
     4681 gtcacgactg gataaatcga aggtcatcaa ctctgctctg gaactgctga atggagtggg
     4741 aattgaagga cttacaaccc gaaagcttgc acaaaaattg ggagttgaac aaccaacact
     4801 ttactggcat gttaagaata agcgagcact tttggatgct cttccaattg aaatgttgga
     4861 tagacatcat actcattcat gtccacttga aggagaatct tggcaagatt tcttgagaaa
     4921 caacgcaaag tcataccgat gcgctctttt gtctcataga gatggagcaa aagttcatct
     4981 tggaacacga ccaactgaaa agcaatacga aacattggaa aaccaacttg ctttcttgtg
     5041 tcaacaagga ttttcattgg aaaatgctct ttacgcattg tctgctgttg gacattttac
     5101 acttggatgc gttttggaag aacaagaaca tcaagttgca aaagaagaac gagaaactcc
     5161 aacaactgat tcaatgccac cacttttgaa gcaagctatc gaacttttcg atagacaagg
     5221 agcagaacca gctttccttt ttggactgga acttattatc tgtggacttg agaaacaact
     5281 gaaatgtgaa tcgggaggag gatctggatc tggatctgga tctggttccg gtcgacaact
     5341 tgagcttccg ccgactgcga ctgccactgc aagtatcatg ccccatgtca tggaacagcc
     5401 cctgtcaacg tcaataaatc cggtcaatga tcggtttaat ggaataccga atcctacccc
     5461 ctacaatagc gatgcggccc tagacgcaat tactcaaacc aatgactacg gatcagtcaa
     5521 tacccatggc attctcagta catatccacc acctgccacc cacttgaacg aggcctcggt
     5581 ggcccttgcg cctggaggtg ctcctcccag accacccccg ccatacgttg atagtacaac
     5641 gaaccatcct ccttaccata gtaacctcgt ccccatggcc aatttcggat attcgactgt
     5701 agactatgat gcgatggtgg atgatctggc atcgatcgag tacacggatg cggttgacgt
     5761 tgatccgcag ttcatgacga acctcgggtt tgtcccaggg tgtaatttta gcgatatcaa
     5821 cacgtatgag caaccatggg gcagtggggc tactaacttc agcctgctga agcaggctgg
     5881 agacgtggag gagaaccctg gacctatggt ctccgagctc attaaagaaa acatgcatat
     5941 gaagctctac atggagggaa ccgtcaacaa ccaccacttc aagtgcacct ccgagggaga
     6001 gggaaagcca tacgagggaa cccaaaccat gcgtatcaag gccgtcgagg gaggaccact
     6061 cccattcgcc ttcgacatcc tcgccacctc cttcatgtac ggatccaagg taagtttaaa
     6121 catatatata ctaactaacc ctgattattt aaattttcag accttcatca accacaccca
     6181 aggaatccca gacttcttca agcaatcctt cccagaggga ttcacctggg agcgtgtcac
     6241 cacctacgag gacggaggag tcctcaccgc cacccaagac acctccctcc aagacggatg
     6301 cctcatctac aacgtcaagg taagtttaaa cagttcggta ctaactaacc atacatattt
     6361 aaattttcag atccgtggag tcaacttccc atccaacgga ccagtcatgc aaaagaagac
     6421 cctcggatgg gaggcctcca ccgagaccct ctacccagcc gacggaggac ttgagggacg
     6481 tgccgacatg gccctcaagc tcgtcggagg aggacacctc atctgcaacc tcaaggtaag
     6541 tttaaacatg attttactaa ctaactaatc tgatttaaat tttcagacca cctaccgttc
     6601 caagaagcca gccaagaacc tcaagatgcc aggagtctac tacgtcgacc gtcgtcttga
     6661 gcgtatcaag gaggccgaca aggagaccta cgtcgagcaa cacgaggtcg ccgtcgcccg
     6721 ttactgcgac ctcccatcca agctcggaca ccgtggatcg ggagagggac gaggaagtct
     6781 tttgacctgc ggagatgttg aggagaaccc aggaccaatg tcgcgtcttg ataaatcgaa
     6841 agttattaat tcagctcttg aacttttgaa cgaagttgga attgaaggat tgacaactag
     6901 aaagcttgca caaaagttgg gagttgaaca accaacactt tattggcatg ttcgaaataa
     6961 gcaaactctt atgaacatgt tgtctgaagc tattcttgca aaacatcata caagatcagc
     7021 tccattgcca actgaatctt ggcaacaatt ccttcaagaa aatgctttgt catttcgaaa
     7081 agcacttttg gttcatagag atggagcacg attgcatatt ggaacatctc caactccacc
     7141 acaattcgaa caagctgaag cacaacttag atgtttgtgc gatgctggat tctcagttga
     7201 agaagcactt ttcatcttgc aatcaatctc tcatttcaca cttggagctg ttttggaaga
     7261 acaagcaact aaccaaatcg aaaacaacca tgttattgat gctgcaccac cacttttgca
     7321 agaagctttc aatattcaag cacgaacatc tgctgaaatg gcattccatt tcggacttaa
     7381 gtcattgatc ttcggatttt ctgctcaact tgatgaaaag aaacatactc caattgaaga
     7441 tggaaacaag ccaaagaaga aaagaaaact gggaagtgga agtaatggaa gtagtggatc
     7501 cagatctgcg ccaccttctt ccgattgtcc gccgccgatt gcctactacc atcatcatcc
     7561 acaacatcag caacaattcc tgccatttcc aatgccatat ttcttggctc caccgccgca
     7621 agctcaacaa ggagctcctt ttccagtgca atatattcca cagcaacatg atttgatgaa
     7681 tagccagcca atgtatgcac caatggcacc gacatactac tatcaaccaa ttaattcgaa
     7741 tggcatgccc atgaattcca actgagcgcc ggtcgctacc attaccaact tgtctggtgt
     7801 caaaaataat aggggccgct gtcatcagag taagtttaaa ctgagttcta ctaactaacg
     7861 agtaatattt aaattttcag catctcgcgc ccgtgcctct gacttctaag tccaattact
     7921 cttcaacatc cctacatgct ctttctccct gtgctcccac cccctatttt tgttattatc
     7981 aaaaaacttc tcttaatttc tttgtttttt agcttctttt aagtcacctc taacaatgaa
     8041 attgtgtaga ttcaaaaata gaattaattc gtaataaaaa gtcgaaaaaa attgtgctcc
     8101 ctccccccat taataataat tctatcccaa aatctacaca atgttctgtg tacacttctt
     8161 atgtttttta cttctgataa atttttttga aacatcatag aaaaaaccgc acacaaaata
     8221 ccttatcata tgttacgttt cagtttatga ccgcaatttt tatttcttcg cacgtctggg
     8281 cctctcatga cgtcaaatca tgctcatcgt gaaaaagttt tggagtattt ttggaatttt
     8341 tcaatcaagt gaaagtttat gaaattaatt ttcctgcttt tgctttttgg ggtttcccct
     8401 attgtttgtc aagatttcga ggacggcgtt tttcttgcta aaatcacaag tattgatgag
     8461 cacgatgcaa gaaagatcgg aagaaggttt gggtttgagg ctcagtggaa ggtgagtaga
     8521 agttgataat ttgaaagtgg agtagtgtct atggggtttt tgccttaaat gacagaatac
     8581 attcccaata taccaaacat aactgtttcc tactagtcgg ccgtacgggc cctttcgtct
     8641 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     8701 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     8761 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     8821 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcggcc
     8881 ttaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     8941 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     9001 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     9061 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     9121 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     9181 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     9241 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     9301 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     9361 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     9421 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     9481 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     9541 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     9601 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     9661 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     9721 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     9781 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     9841 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     9901 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     9961 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
    10021 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
    10081 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
    10141 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
    10201 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
    10261 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
    10321 ctctgctaat cc
//
