LOCUS       sequence-329943-        7370 bp    DNA     circular SYN 14-APR-2025
DEFINITION  TRE::nCre, inducible nCre expression when co-express with rtTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7370)
  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 7370)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: TC872
FEATURES             Location/Qualifiers
     source          1..7370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      77..665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     566..585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1074..1090
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1074..1090
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1156..1174
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1192..1210
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1228..1246
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1264..1282
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1300..1318
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1336..1354
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1372..1390
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1674..1694
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1695..2723
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     terminator      2724..3047
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(3160..3177)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3160..3176)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3998..4792
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4052..4071)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4662..4681
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5493..5509
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5494..5510
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6165..6187
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6225..6243)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6311..6415
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6416..6484,6485..7276)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6634..6653)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggt cttttgctgg ccttttgctc
      721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      781 gagctgatac cgcaccgtac gtctcgagga attctatcag gtgtacaagt atgaaatgtc
      841 gtttcgaaca tatagatgtc tcgcaaacgt aaatatttat cgattgtcat aaaactttga
      901 ccttgtgaag tgtcaacctt gactgtcgaa ccaccatagt ttggcgcgaa ttgagcgtca
      961 taattgttta ctctcagtgc agtcaacatg tcgagtttcg tgccgaataa agagcaaacg
     1021 cggacagtat taattttctg ttttcatttg aagaaaacag ctgcggccgg atccaggaaa
     1081 cagctatgac catgattacg ccaagctacg taatacgact cactagtggg cagatcttcg
     1141 aatgcatgag tttactccct atcagtgata gagaacgtat gaagagttta ctccctatca
     1201 gtgatagaga acgtatgcag actttactcc ctatcagtga tagagaacgt ataaggagtt
     1261 tactccctat cagtgataga gaacgtatga ccagtttact ccctatcagt gatagagaac
     1321 gtatctacag tttactccct atcagtgata gagaacgtat atccagttta ctccctatca
     1381 gtgatagaga acgtatgtcg aggtaggcgt gatcgatttt ttgcaaatta cgagcgttgt
     1441 agggggcgga gcgataggtc ctataggttt tggtatatca tcattcattc attcattggt
     1501 acattcattt acccaccttc ctctttctga gcttctctgg agttctgtgc ttcctttttc
     1561 ccttatcttt atactgtaat ttttaacttt cagaaggacc caaaggtatg tttcgaatga
     1621 tactaacata acatagaaca ttttcaggag attggagaga caccatgggc gcacccaaga
     1681 agaagaggaa ggtgtccaat ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg
     1741 atgcaacgag tgatgaggtt cgcaagaacc tgatggacat gttcagggat cgccaggcgt
     1801 tttctgagca tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca
     1861 agttgaataa ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat tatcttctat
     1921 atcttcaggc gcgcggtctg gcagtaaaaa ctatccagca acatttgggc cagctaaaca
     1981 tgcttcatcg tcggtccggg ctgccacgac caagtgacag caatgctgtt tcactggtta
     2041 tgcggcggat ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag gctctagcgt
     2101 tcgaacgcac tgatttcgac caggttcgtt cactcatgga aaatagcgat cgctgccagg
     2161 atatacgtaa tctggcattt ctggggattg cttataacac cctgttacgt atagccgaaa
     2221 ttgccaggat cagggttaaa gatatctcac gtactgacgg tgggagaatg ttaatccata
     2281 ttggcagaac gaaaacgctg gttagcaccg caggtgtaga gaaggcactt agcctggggg
     2341 taactaaact ggtcgagcga tggatttccg tctctggtgt agctgatgat ccgaataact
     2401 acctgttttg ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc agccagctat
     2461 caactcgcgc cctggaaggg atttttgaag caactcatcg attgatttac ggcgctaagg
     2521 atgactctgg tcagagatac ctggcctggt ctggacacag tgcccgtgtc ggagccgcgc
     2581 gagatatggc ccgcgctgga gtttcaatac cggagatcat gcaagctggt ggctggacca
     2641 atgtaaatat tgtcatgaac tatatccgta acctggatag tgaaacaggg gcaatggtgc
     2701 gcctgctgga agatggcgat tagatgcaag atcctttcaa gcattccctt cttctctatc
     2761 actcttcttt ctttttgtca aaaaattctc tcgctaattt atttgctttt ttaatgttat
     2821 tattttatga ctttttatag tcactgaaaa gtttgcatct gagtgaagtg aatgctatca
     2881 aaatgtgatt ctgtctgatg tactttcaca atctctcttc aattccattt tgaagtgctt
     2941 taaacccgaa aggttgagaa aaatgcgagc gctcaaatat ttgtattgtg ttcgttgagt
     3001 gacccaacaa aaagaggaaa ctttattgtg ccgccaagaa aaaagtctcc tgcaggatat
     3061 ctggatccac gaagcttccc atggtgacgt caccggttct agatacctag gtgagctctg
     3121 gtaccctcta gtcaaggcct taagtgagtc gtattacgga ctggccgtcg ttttacaggc
     3181 cggatgggcc atggcacctt aacatttcta aattttagtt taataaaaaa attttaaaat
     3241 aatacgagtt ttgagactaa ctttcaataa aaaccaaaaa attgcttcaa aaaagagctc
     3301 gcgccgattt acgataaaca cagcgcgcta cgacactccg aaaaccgcgc cgcaccatta
     3361 cattctccgc cccaccgcac ggtttgcgga aggcgatgaa attgctgggc ctaaaatttt
     3421 aacccttttt ttcccgtttt tcaactgttt tcccgatttt ttcgatattt ttttcaggtt
     3481 tttagttaga aaagtatata aaaattgcat tatttgtagt tttcgctcgt aatttgtgat
     3541 agaaaacggc caatttgcct agatttgtga ttctaccaag tggaattttt gggaaaatat
     3601 caatattcca cgatttttat tcaaatcata acgtttcgaa attatcagct taaataaaac
     3661 taaaataaat aataaaatac taaaaaccct gaatttctcg gttctcggtt cagcttgagt
     3721 tcgttctata cacgttatcg atttttgtca tttcgtttaa tcatgggttt tgattcattt
     3781 taaatatgaa atttgattta ttcctcgttt cgaagttggt ttgtcttgtc cggccggtaa
     3841 acaaatgtta ttaaaaaaaa taataaattt cgaaattttt cgtaatgttc cgcaatcgtg
     3901 tttttccgtt tatctcgtct tcccttcctt caaatatact atttattttc tcaattcaca
     3961 tattttcagc cagccctgct caacaagtgg aataaaaatg attgaacaag atggattgca
     4021 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4081 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4141 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4201 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4261 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4321 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4381 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4441 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcaccagc
     4501 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4561 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4621 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4681 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4741 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagtccaatt
     4801 actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt
     4861 atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagccac ctctaacaat
     4921 gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc
     4981 tccctccccc cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt
     5041 cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac cgcacacaaa
     5101 ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct
     5161 gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat
     5221 ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt tggggtttcc
     5281 cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac aagtattgat
     5341 gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt
     5401 agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa
     5461 tacattccca atataccaaa cataactgtt tctcgaggtc gacggtatcg ataagcttga
     5521 tccggatctg cggcttactc accagacctg gccccatccg attaccacct attcgcttcg
     5581 atgggacacg cactcgctga gcagcgcttc gattcttacg aaagtgtgaa aaaatggctc
     5641 gatgaatggt tcgccgcaaa agacgatgag ttctactggc gtggaatcca caaattgccc
     5701 gagagatggg aaaaatgtgt agctagcgac ggcaaatact ttgaataaat gattttttct
     5761 ttttccacaa aatttaacgt gttttttgat ttaaaaaaaa cgacatttca tacttgtaca
     5821 cctgatatct agatacctag gtgagctctg gtacccccct ggcgttaccc aacttaatcg
     5881 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     5941 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     6001 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtgcaa tctgctctga
     6061 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     6121 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     6181 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     6241 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     6301 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6361 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6421 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6481 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6541 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6601 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6661 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6721 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6781 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6841 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6901 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6961 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     7021 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     7081 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     7141 tatcattgca gcactggggt cagatggtaa gccctcccgt atcgtagtta tctacacgac
     7201 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     7261 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     7321 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
//
