LOCUS       sequence-244009-        8034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prpl-28::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             1720..1776
                     /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             2635..2688
                     /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(4539..4561)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4661..4680
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4758..4776)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4844..4948
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4949..5017,5018..5809)
                     /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(5167..5186)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5980..6568
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6469..6488
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6722..6739
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6856..6877
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6892..6897,6898..6915,6916..6922)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6930..6946
                     /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     6935..6957
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6954..6970
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6954..6970
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttaataaaat tatataattt tgaataaaat caaattaaaa ataagattaa ttgaaacaca
       61 acaatagaat gcagaaaatt actcataaga aataatttat tttgaaagaa gttgtgtcga
      121 tttacgataa aaactacacg caaacgaggc tccgcgttta caccgtttgg caatggagcg
      181 cgcttgctat tagattttag gccagctcaa cgaaagttta atttattttt cttctttttt
      241 tccatttttt tgagtaaaat atcgaatttt ttgtattaat tctgtaaatt ttataatttg
      301 caactatttt ggcgtattct gcttgttttt atcagcgaat ttttatgcat ttcagtcaaa
      361 agtttgagca ggaaaaatga catttaggac tttgtaaaat tgtattctta aatttgcttg
      421 aaaacctacg tgtacaattg attttattga atttttctta cgtttcccgt tgtatcttat
      481 tgtatttaac tttcatcata aaataattta tttccaggta aaatatccga cgctctcgtg
      541 gatcaaaaaa tgtcacgact ggataaatcg aaggtcatca actctgctct ggaactgctg
      601 aatggagtgg gaattgaagg acttacaacc cgaaagcttg cacaaaaatt gggagttgaa
      661 caaccaacac tttactggca tgttaagaat aagcgagcac ttttggatgc tcttccaatt
      721 gaaatgttgg atagacatca tactcattca tgtccacttg aaggagaatc ttggcaagat
      781 ttcttgagaa acaacgcaaa gtcataccga tgcgctcttt tgtctcatag agatggagca
      841 aaagttcatc ttggaacacg accaactgaa aagcaatacg aaacattgga aaaccaactt
      901 gctttcttgt gtcaacaagg attttcattg gaaaatgctc tttacgcatt gtctgctgtt
      961 ggacatttta cacttggatg cgttttggaa gaacaagaac atcaagttgc aaaagaagaa
     1021 cgagaaactc caacaactga ttcaatgcca ccacttttga agcaagctat cgaacttttc
     1081 gatagacaag gagcagaacc agctttcctt tttggactgg aacttattat ctgtggactt
     1141 gagaaacaac tgaaatgtga atcgggagga ggatctggat ctggatctgg atctggttcc
     1201 ggtcgacaac ttgagcttcc gccgactgcg actgccactg caagtatcat gccccatgtc
     1261 atggaacagc ccctgtcaac gtcaataaat ccggtcaatg atcggtttaa tggaataccg
     1321 aatcctaccc cctacaatag cgatgcggcc ctagacgcaa ttactcaaac caatgactac
     1381 ggatcagtca atacccatgg cattctcagt acatatccac cacctgccac ccacttgaac
     1441 gaggcctcgg tggcccttgc gcctggaggt gctcctccca gaccaccccc gccatacgtt
     1501 gatagtacaa cgaaccatcc tccttaccat agtaacctcg tccccatggc caatttcgga
     1561 tattcgactg tagactatga tgcgatggtg gatgatctgg catcgatcga gtacacggat
     1621 gcggttgacg ttgatccgca gttcatgacg aacctcgggt ttgtcccagg gtgtaatttt
     1681 agcgatatca acacgtatga gcaaccatgg ggcagtgggg ctactaactt cagcctgctg
     1741 aagcaggctg gagacgtgga ggagaaccct ggacctatgg tctccgagct cattaaagaa
     1801 aacatgcata tgaagctcta catggaggga accgtcaaca accaccactt caagtgcacc
     1861 tccgagggag agggaaagcc atacgaggga acccaaacca tgcgtatcaa ggccgtcgag
     1921 ggaggaccac tcccattcgc cttcgacatc ctcgccacct ccttcatgta cggatccaag
     1981 gtaagtttaa acatatatat actaactaac cctgattatt taaattttca gaccttcatc
     2041 aaccacaccc aaggaatccc agacttcttc aagcaatcct tcccagaggg attcacctgg
     2101 gagcgtgtca ccacctacga ggacggagga gtcctcaccg ccacccaaga cacctccctc
     2161 caagacggat gcctcatcta caacgtcaag gtaagtttaa acagttcggt actaactaac
     2221 catacatatt taaattttca gatccgtgga gtcaacttcc catccaacgg accagtcatg
     2281 caaaagaaga ccctcggatg ggaggcctcc accgagaccc tctacccagc cgacggagga
     2341 cttgagggac gtgccgacat ggccctcaag ctcgtcggag gaggacacct catctgcaac
     2401 ctcaaggtaa gtttaaacat gattttacta actaactaat ctgatttaaa ttttcagacc
     2461 acctaccgtt ccaagaagcc agccaagaac ctcaagatgc caggagtcta ctacgtcgac
     2521 cgtcgtcttg agcgtatcaa ggaggccgac aaggagacct acgtcgagca acacgaggtc
     2581 gccgtcgccc gttactgcga cctcccatcc aagctcggac accgtggatc gggagaggga
     2641 cgaggaagtc ttttgacctg cggagatgtt gaggagaacc caggaccaat gtcgcgtctt
     2701 gataaatcga aagttattaa ttcagctctt gaacttttga acgaagttgg aattgaagga
     2761 ttgacaacta gaaagcttgc acaaaagttg ggagttgaac aaccaacact ttattggcat
     2821 gttcgaaata agcaaactct tatgaacatg ttgtctgaag ctattcttgc aaaacatcat
     2881 acaagatcag ctccattgcc aactgaatct tggcaacaat tccttcaaga aaatgctttg
     2941 tcatttcgaa aagcactttt ggttcataga gatggagcac gattgcatat tggaacatct
     3001 ccaactccac cacaattcga acaagctgaa gcacaactta gatgtttgtg cgatgctgga
     3061 ttctcagttg aagaagcact tttcatcttg caatcaatct ctcatttcac acttggagct
     3121 gttttggaag aacaagcaac taaccaaatc gaaaacaacc atgttattga tgctgcacca
     3181 ccacttttgc aagaagcttt caatattcaa gcacgaacat ctgctgaaat ggcattccat
     3241 ttcggactta agtcattgat cttcggattt tctgctcaac ttgatgaaaa gaaacatact
     3301 ccaattgaag atggaaacaa gccaaagaag aaaagaaaac tgggaagtgg aagtaatgga
     3361 agtagtggat ccagatctgc gccaccttct tccgattgtc cgccgccgat tgcctactac
     3421 catcatcatc cacaacatca gcaacaattc ctgccatttc caatgccata tttcttggct
     3481 ccaccgccgc aagctcaaca aggagctcct tttccagtgc aatatattcc acagcaacat
     3541 gatttgatga atagccagcc aatgtatgca ccaatggcac cgacatacta ctatcaacca
     3601 attaattcga atggcatgcc catgaattcc aactgagcgc cggtcgctac cattaccaac
     3661 ttgtctggtg tcaaaaataa taggggccgc tgtcatcaga gtaagtttaa actgagttct
     3721 actaactaac gagtaatatt taaattttca gcatctcgcg cccgtgcctc tgacttctaa
     3781 gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca ccccctattt
     3841 ttgttattat caaaaaactt ctcttaattt ctttgttttt tagcttcttt taagtcacct
     3901 ctaacaatga aattgtgtag attcaaaaat agaattaatt cgtaataaaa agtcgaaaaa
     3961 aattgtgctc cctcccccca ttaataataa ttctatccca aaatctacac aatgttctgt
     4021 gtacacttct tatgtttttt acttctgata aatttttttg aaacatcata gaaaaaaccg
     4081 cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc
     4141 gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt
     4201 tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg
     4261 gggtttcccc tattgtttgt caagatttcg aggacggcgt ttttcttgct aaaatcacaa
     4321 gtattgatga gcacgatgca agaaagatcg gaagaaggtt tgggtttgag gctcagtgga
     4381 aggtgagtag aagttgataa tttgaaagtg gagtagtgtc tatggggttt ttgccttaaa
     4441 tgacagaata cattcccaat ataccaaaca taactgtttc ctactagtcg gccgtacggg
     4501 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     4561 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4621 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     4681 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     4741 atcaggcggc cttaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4801 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4861 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4921 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4981 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5041 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5101 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5161 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5221 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5281 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5341 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5401 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5461 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5521 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5581 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5641 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5701 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5761 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5821 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5881 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5941 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6001 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6061 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6121 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6181 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6241 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6301 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6361 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6421 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6481 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6541 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6601 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6661 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6721 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6781 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6841 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     6901 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6961 cagctatgac catgattacg ccaagctgta agtttaaaca tgatcttact aactaactat
     7021 tctcatttaa attttcagag cttaaaaatg gctgaaatca ctcacaacga tggatacgct
     7081 aacaacttgg aaatgaaata agcttgcatg cctgcagttt gtgcaacaaa ttgaggaagg
     7141 cattctagag aagaatttcg gcatttcgca atgatacaaa atgaagagaa taaggaaatg
     7201 acggtgaaat aaccggtttg aaggtaaaaa tgataaagta gacaattgaa tttttctagt
     7261 tcattctcca acaataaatt cacaacaaaa caaatctgcc tagataaaaa actaaaagat
     7321 atatcaagaa tacatctaga atcctttgct gtagaagaat tgctcaataa gaacacgtgg
     7381 agctccttta attttgatac gacctttcaa ctcatcgacg tgactggcga tcgaatgtcc
     7441 aagatgttct tcaagatagg agcggagatc attttcgcaa gcctgaaaga tatttatgtt
     7501 gattcaggat ttaataatgt tcccaactta caaaaatgtc tccgtcgacg tttctgatag
     7561 ttactagctc gacataatca aaatccaacg ttttctcatt cagtaaatcc ctttttcgtt
     7621 ccaaataaag aggaagaaga tggtcatgac gacgtcgaat gtagtattga tgtgtcttgg
     7681 ctgcttctga aatatttcga tgcggtatga atttaatctg tatatacctg tcggtggtgt
     7741 ccatccgctt ggtgtaggat acttttcgtg ctcaggaaca ttcggcacca cttctattgg
     7801 catcaatctt tccacatacg accaatcgat aggagcttct tcgaccgttg caagacttct
     7861 ggctgtaatt gctagtagaa aaattatatg ataataaata acttacactt ttcagggaga
     7921 gcatgtttcc atggattttc ccaaagtttc tcagcttttt cgactgaagt tgttgaaaaa
     7981 gacaacctgg aaactgctcg aggggagaaa atgcggagag aagaacccag catc
//
