LOCUS       sequence-325965-        8034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prpl28::rtetR-QFAD::p2A::mKate::T2A::tetR-pie1, .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8034)
  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 8034)
  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: TC374
FEATURES             Location/Qualifiers
     source          1..8034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2655..2711
                     /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             3570..3623
                     /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(5474..5496)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5596..5615
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5693..5711)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5779..5883
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5884..5952,5953..6744)
                     /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(6102..6121)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6915..7503
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7404..7423
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7657..7674
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7791..7812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7827..7832,7833..7850,7851..7857)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7865..7881
                     /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     7870..7892
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7889..7905
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7889..7905
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcagtt tgtgcaacaa attgaggaag gcattctaga gaagaatttc
       61 ggcatttcgc aatgatacaa aatgaagaga ataaggaaat gacggtgaaa taaccggttt
      121 gaaggtaaaa atgataaagt agacaattga atttttctag ttcattctcc aacaataaat
      181 tcacaacaaa acaaatctgc ctagataaaa aactaaaaga tatatcaaga atacatctag
      241 aatcctttgc tgtagaagaa ttgctcaata agaacacgtg gagctccttt aattttgata
      301 cgacctttca actcatcgac gtgactggcg atcgaatgtc caagatgttc ttcaagatag
      361 gagcggagat cattttcgca agcctgaaag atatttatgt tgattcagga tttaataatg
      421 ttcccaactt acaaaaatgt ctccgtcgac gtttctgata gttactagct cgacataatc
      481 aaaatccaac gttttctcat tcagtaaatc cctttttcgt tccaaataaa gaggaagaag
      541 atggtcatga cgacgtcgaa tgtagtattg atgtgtcttg gctgcttctg aaatatttcg
      601 atgcggtatg aatttaatct gtatatacct gtcggtggtg tccatccgct tggtgtagga
      661 tacttttcgt gctcaggaac attcggcacc acttctattg gcatcaatct ttccacatac
      721 gaccaatcga taggagcttc ttcgaccgtt gcaagacttc tggctgtaat tgctagtaga
      781 aaaattatat gataataaat aacttacact tttcagggag agcatgtttc catggatttt
      841 cccaaagttt ctcagctttt tcgactgaag ttgttgaaaa agacaacctg gaaactgctc
      901 gaggggagaa aatgcggaga gaagaaccca gcatcttaat aaaattatat aattttgaat
      961 aaaatcaaat taaaaataag attaattgaa acacaacaat agaatgcaga aaattactca
     1021 taagaaataa tttattttga aagaagttgt gtcgatttac gataaaaact acacgcaaac
     1081 gaggctccgc gtttacaccg tttggcaatg gagcgcgctt gctattagat tttaggccag
     1141 ctcaacgaaa gtttaattta tttttcttct ttttttccat ttttttgagt aaaatatcga
     1201 attttttgta ttaattctgt aaattttata atttgcaact attttggcgt attctgcttg
     1261 tttttatcag cgaattttta tgcatttcag tcaaaagttt gagcaggaaa aatgacattt
     1321 aggactttgt aaaattgtat tcttaaattt gcttgaaaac ctacgtgtac aattgatttt
     1381 attgaatttt tcttacgttt cccgttgtat cttattgtat ttaactttca tcataaaata
     1441 atttatttcc aggtaaaata tccgacgctc tcgtggatca aaaaatgtca cgactggata
     1501 aatcgaaggt catcaactct gctctggaac tgctgaatgg agtgggaatt gaaggactta
     1561 caacccgaaa gcttgcacaa aaattgggag ttgaacaacc aacactttac tggcatgtta
     1621 agaataagcg agcacttttg gatgctcttc caattgaaat gttggataga catcatactc
     1681 attcatgtcc acttgaagga gaatcttggc aagatttctt gagaaacaac gcaaagtcat
     1741 accgatgcgc tcttttgtct catagagatg gagcaaaagt tcatcttgga acacgaccaa
     1801 ctgaaaagca atacgaaaca ttggaaaacc aacttgcttt cttgtgtcaa caaggatttt
     1861 cattggaaaa tgctctttac gcattgtctg ctgttggaca ttttacactt ggatgcgttt
     1921 tggaagaaca agaacatcaa gttgcaaaag aagaacgaga aactccaaca actgattcaa
     1981 tgccaccact tttgaagcaa gctatcgaac ttttcgatag acaaggagca gaaccagctt
     2041 tcctttttgg actggaactt attatctgtg gacttgagaa acaactgaaa tgtgaatcgg
     2101 gaggaggatc tggatctgga tctggatctg gttccggtcg acaacttgag cttccgccga
     2161 ctgcgactgc cactgcaagt atcatgcccc atgtcatgga acagcccctg tcaacgtcaa
     2221 taaatccggt caatgatcgg tttaatggaa taccgaatcc taccccctac aatagcgatg
     2281 cggccctaga cgcaattact caaaccaatg actacggatc agtcaatacc catggcattc
     2341 tcagtacata tccaccacct gccacccact tgaacgaggc ctcggtggcc cttgcgcctg
     2401 gaggtgctcc tcccagacca cccccgccat acgttgatag tacaacgaac catcctcctt
     2461 accatagtaa cctcgtcccc atggccaatt tcggatattc gactgtagac tatgatgcga
     2521 tggtggatga tctggcatcg atcgagtaca cggatgcggt tgacgttgat ccgcagttca
     2581 tgacgaacct cgggtttgtc ccagggtgta attttagcga tatcaacacg tatgagcaac
     2641 catggggcag tggggctact aacttcagcc tgctgaagca ggctggagac gtggaggaga
     2701 accctggacc tatggtctcc gagctcatta aagaaaacat gcatatgaag ctctacatgg
     2761 agggaaccgt caacaaccac cacttcaagt gcacctccga gggagaggga aagccatacg
     2821 agggaaccca aaccatgcgt atcaaggccg tcgagggagg accactccca ttcgccttcg
     2881 acatcctcgc cacctccttc atgtacggat ccaaggtaag tttaaacata tatatactaa
     2941 ctaaccctga ttatttaaat tttcagacct tcatcaacca cacccaagga atcccagact
     3001 tcttcaagca atccttccca gagggattca cctgggagcg tgtcaccacc tacgaggacg
     3061 gaggagtcct caccgccacc caagacacct ccctccaaga cggatgcctc atctacaacg
     3121 tcaaggtaag tttaaacagt tcggtactaa ctaaccatac atatttaaat tttcagatcc
     3181 gtggagtcaa cttcccatcc aacggaccag tcatgcaaaa gaagaccctc ggatgggagg
     3241 cctccaccga gaccctctac ccagccgacg gaggacttga gggacgtgcc gacatggccc
     3301 tcaagctcgt cggaggagga cacctcatct gcaacctcaa ggtaagttta aacatgattt
     3361 tactaactaa ctaatctgat ttaaattttc agaccaccta ccgttccaag aagccagcca
     3421 agaacctcaa gatgccagga gtctactacg tcgaccgtcg tcttgagcgt atcaaggagg
     3481 ccgacaagga gacctacgtc gagcaacacg aggtcgccgt cgcccgttac tgcgacctcc
     3541 catccaagct cggacaccgt ggatcgggag agggacgagg aagtcttttg acctgcggag
     3601 atgttgagga gaacccagga ccaatgtcgc gtcttgataa atcgaaagtt attaattcag
     3661 ctcttgaact tttgaacgaa gttggaattg aaggattgac aactagaaag cttgcacaaa
     3721 agttgggagt tgaacaacca acactttatt ggcatgttcg aaataagcaa actcttatga
     3781 acatgttgtc tgaagctatt cttgcaaaac atcatacaag atcagctcca ttgccaactg
     3841 aatcttggca acaattcctt caagaaaatg ctttgtcatt tcgaaaagca cttttggttc
     3901 atagagatgg agcacgattg catattggaa catctccaac tccaccacaa ttcgaacaag
     3961 ctgaagcaca acttagatgt ttgtgcgatg ctggattctc agttgaagaa gcacttttca
     4021 tcttgcaatc aatctctcat ttcacacttg gagctgtttt ggaagaacaa gcaactaacc
     4081 aaatcgaaaa caaccatgtt attgatgctg caccaccact tttgcaagaa gctttcaata
     4141 ttcaagcacg aacatctgct gaaatggcat tccatttcgg acttaagtca ttgatcttcg
     4201 gattttctgc tcaacttgat gaaaagaaac atactccaat tgaagatgga aacaagccaa
     4261 agaagaaaag aaaactggga agtggaagta atggaagtag tggatccaga tctgcgccac
     4321 cttcttccga ttgtccgccg ccgattgcct actaccatca tcatccacaa catcagcaac
     4381 aattcctgcc atttccaatg ccatatttct tggctccacc gccgcaagct caacaaggag
     4441 ctccttttcc agtgcaatat attccacagc aacatgattt gatgaatagc cagccaatgt
     4501 atgcaccaat ggcaccgaca tactactatc aaccaattaa ttcgaatggc atgcccatga
     4561 attccaactg agcgccggtc gctaccatta ccaacttgtc tggtgtcaaa aataataggg
     4621 gccgctgtca tcagagtaag tttaaactga gttctactaa ctaacgagta atatttaaat
     4681 tttcagcatc tcgcgcccgt gcctctgact tctaagtcca attactcttc aacatcccta
     4741 catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa aacttctctt
     4801 aatttctttg ttttttagct tcttttaagt cacctctaac aatgaaattg tgtagattca
     4861 aaaatagaat taattcgtaa taaaaagtcg aaaaaaattg tgctccctcc ccccattaat
     4921 aataattcta tcccaaaatc tacacaatgt tctgtgtaca cttcttatgt tttttacttc
     4981 tgataaattt ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt atcatatgtt
     5041 acgtttcagt ttatgaccgc aatttttatt tcttcgcacg tctgggcctc tcatgacgtc
     5101 aaatcatgct catcgtgaaa aagttttgga gtatttttgg aatttttcaa tcaagtgaaa
     5161 gtttatgaaa ttaattttcc tgcttttgct ttttggggtt tcccctattg tttgtcaaga
     5221 tttcgaggac ggcgtttttc ttgctaaaat cacaagtatt gatgagcacg atgcaagaaa
     5281 gatcggaaga aggtttgggt ttgaggctca gtggaaggtg agtagaagtt gataatttga
     5341 aagtggagta gtgtctatgg ggtttttgcc ttaaatgaca gaatacattc ccaatatacc
     5401 aaacataact gtttcctact agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg
     5461 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     5521 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     5581 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     5641 aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa gggcctcgtg
     5701 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5761 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5821 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5881 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5941 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6001 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6061 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6121 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6181 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6241 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6301 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6361 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6421 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6481 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6541 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6601 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6661 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6721 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6781 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6841 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6901 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6961 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7021 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7081 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7141 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7201 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7261 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7321 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7381 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7441 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7501 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7561 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7621 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7681 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7741 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7801 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7861 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7921 ctgtaagttt aaacatgatc ttactaacta actattctca tttaaatttt cagagcttaa
     7981 aaatggctga aatcactcac aacgatggat acgctaacaa cttggaaatg aaat
//
