LOCUS       sequence-325963-        4836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRE::GFP, inducible GFP expression when co-express with rtTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4836)
  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 4836)
  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: TC358
FEATURES             Location/Qualifiers
     source          1..4836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    118..136
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    154..172
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    190..208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    226..244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    262..280
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    298..316
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    334..352
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(598..626)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1374..1394
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2361..2383)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2483..2502
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2580..2598)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2666..2770
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2771..2839,2840..3631)
                     /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(2989..3008)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3802..4390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4291..4310
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4544..4561
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4678..4699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4714..4719,4720..4737,4738..4744)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4752..4768
                     /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     4757..4779
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4776..4792
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4776..4792
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aactattctc atttaaattt tcagagctta aaaatggctg aaatcactca caacgatgga
       61 tacgctaaca acttggaaat gaaataagct tgcatgcccg tcttcactcg agtttactcc
      121 ctatcagtga tagagaacgt atgaagagtt tactccctat cagtgataga gaacgtatgc
      181 agactttact ccctatcagt gatagagaac gtataaggag tttactccct atcagtgata
      241 gagaacgtat gaccagttta ctccctatca gtgatagaga acgtatctac agtttactcc
      301 ctatcagtga tagagaacgt atatccagtt tactccctat cagtgataga gaacgtatgt
      361 cgaggtaggc gtgatcgatt ttttgcaaat tacgagcgtt gtagggggcg gagcgatagg
      421 tcctataggt tttggtatat catcattcat tcattcattg gtacattcat ttacccacct
      481 tcctctttct gagcttctct ggagttctgt gcttcctttt tcccttatct ttatactgta
      541 atttttaact ttcagccggt agaaaaaatg agtaaaggag aagaactttt cactggagtt
      601 gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga
      661 gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga
      721 aaactacctg ttccatgggt aagtttaaac atatatatac taactaaccc tgattattta
      781 aattttcagc caacacttgt cactactttc tgttatggtg ttcaatgctt ctcgagatac
      841 ccagatcata tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
      901 gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtaagtt taaacagttc
      961 ggtactaact aaccatacat atttaaattt tcaggtgctg aagtcaagtt tgaaggtgat
     1021 acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt
     1081 ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa
     1141 aagaatggaa tcaaagttgt aagtttaaac atgattttac taactaacta atctgattta
     1201 aattttcaga acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
     1261 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     1321 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
     1381 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caaatagcat
     1441 tcgtagaatt ccaactgagc gccggtcgct accattacca acttgtctgg tgtcaaaaat
     1501 aataggggcc gctgtcatca gagtaagttt aaactgagtt ctactaacta acgagtaata
     1561 tttaaatttt cagcatctcg cgcccgtgcc tctgacttct aagtccaatt actcttcaac
     1621 atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt atcaaaaaac
     1681 ttctcttaat ttctttgttt tttagcttct tttaagtcac ctctaacaat gaaattgtgt
     1741 agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc
     1801 cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt cttatgtttt
     1861 ttacttctga taaatttttt tgaaacatca tagaaaaaac cgcacacaaa ataccttatc
     1921 atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct gggcctctca
     1981 tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat ttttcaatca
     2041 agtgaaagtt tatgaaatta attttcctgc ttttgctttt tggggtttcc cctattgttt
     2101 gtcaagattt cgaggacggc gtttttcttg ctaaaatcac aagtattgat gagcacgatg
     2161 caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt agaagttgat
     2221 aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa tacattccca
     2281 atataccaaa cataactgtt tcctactagt cggccgtacg ggccctttcg tctcgcgcgt
     2341 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     2401 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     2461 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     2521 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg gccttaaggg
     2581 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2641 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2701 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2761 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2821 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2881 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2941 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3001 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3061 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3121 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3181 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3241 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3301 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3361 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3421 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3481 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3541 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3601 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3661 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3721 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3781 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3841 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3901 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3961 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4021 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4081 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4141 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4201 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4261 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4321 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4381 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4441 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4501 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4561 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4621 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     4681 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     4741 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     4801 cgccaagctg taagtttaaa catgatctta ctaact
//
