LOCUS       sequence-325434-        5895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Introduce Dox-inducible (TRE promoter) GFP cDNA into (ES) cells by 
            recombination-mediated cassette exchange.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5895)
  TITLE     Karreth lab plasmids
REFERENCE   2  (bases 1 to 5895)
  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"
COMMENT     Sequence Label: cTRE-GFP
FEATURES             Location/Qualifiers
     source          1..5895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(25..90,91..128)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     polyA_signal    297..431
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    551..685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    join(697..715,716..738,739..757,758..780,781..799,800..822,
                     823..841,842..864,865..883,884..906,907..925,926..948,
                     949..967)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        1000..1038
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1035..1059
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(1144..1146,1147..1149,1150..1863)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANLKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1189..1210)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1450..1469)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1797..1818
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          1924..2496
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     rep_origin      complement(2824..3412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2904..2923)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3583..4374,4375..4443))
                     /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     4206..4225
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4444..4548)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4574..5029)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4711..4732)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4923..4942
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5156..5178
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5170..5187
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5171..5187
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5194..5213
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5194..5212
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5245..5261
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5245..5261
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        5305..5804
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5340..5363)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5737..5756
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    join(5850..5895,1..2)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
        1 tcgatccccc gggctgcaga tctgtagggc gcagtagtcc agggtttcct tgatgatgtc
       61 atacttatcc tgtccctttt ttttccacag ctcgcggttg aggacaaact cttcgcggtc
      121 tttccagtgg ggatcgacgg tatcgataag cttgcagatc tgcgactcta gaggatctgc
      181 gactctagag gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
      241 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
      301 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
      361 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
      421 atgtctggat ctgcgactct agaggatcat aatcagccat accacatttg tagaggtttt
      481 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
      541 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
      601 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
      661 caatgtatct tatcatgtct ggatcgagtt taccactccc tatcagtgat agagaaaagt
      721 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
      781 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
      841 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
      901 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
      961 atagagaaaa gtgaaagtcg agctcggtac ccgggtcgag gtaggcgtgt acggtgggag
     1021 gcctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
     1081 tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggccc cgaattgacc
     1141 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1201 gacggcgacg taaacggcca caagttcagc gtgtctggcg agggcgaggg cgatgccacc
     1261 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1321 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     1381 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1441 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1501 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1561 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     1621 aacggcatca aggcgaacct caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1681 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1741 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1801 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1861 taatgacgta cggcggccgc ctcgagctgc tagaattcca attgacgcgt tgagaacttc
     1921 agggtgagtt tggggaccct tgattgttct ttctttttcg ctattgtaaa attcatgtta
     1981 tatggagggg gcaaagtttt cagggtgttg tttagaatgg gaagatgtcc cttgtatcac
     2041 catggaccct catgataatt ttgtttcttt cactttctac tctgttgaca accattgtct
     2101 cctcttattt tcttttcatt ttctgtaact ttttcgttaa actttagctt gcatttgtaa
     2161 cgaattttta aattcacttt tgtttatttg tcagattgta agtactttct ctaatcactt
     2221 ttttttcaag gcaatcaggg tatattatat tgtacttcag cacagtttta gagaacaatt
     2281 gttataatta aatgataagg tagaatattt ctgcatataa attctggctg gcgtggaaat
     2341 attcttattg gtagaaacaa ctacaccctg gtcatcatcc tgcctttctc tttatggtta
     2401 caatgatata cactgtttga gatgaggata aaatactctg agtccaaacc gggcccctct
     2461 gctaaccatg ttcatgcctt cttctctttc ctacagctcc tgggcaacgt gctggttgtt
     2521 gtgctgtctc atcattttgg caaaggattc actcctcagg tgcaggctgc ctatcagaag
     2581 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatcg ttttccctct
     2641 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     2701 aaatttattt tcattgcaat agtgtgttgg aatttctcga tcgctagcat cctaggcgcg
     2761 ccacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2821 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2881 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2941 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3001 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3061 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3121 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3181 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3241 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3301 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3361 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3421 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3481 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3541 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3601 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3661 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3721 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3781 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3841 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3901 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3961 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4021 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4081 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4141 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4201 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4261 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4321 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4381 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4441 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4501 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4561 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     4621 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4681 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4741 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     4801 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     4861 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4921 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4981 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     5041 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     5101 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     5161 tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg
     5221 gagctccacc gcggtggcgg ccgctctaga actagtggat cccccgggct gcaggaatta
     5281 attcgtcgac ctcgaaattc taccgggtag gggaggcgct tttcccaagg cagtctggag
     5341 catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca
     5401 cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc
     5461 caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca
     5521 ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg
     5581 agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt
     5641 tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg
     5701 gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca
     5761 cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc atccatctag
     5821 atctcgatcg agcagctgaa gcttatggag aagttactat tccgaagttc ctattctcta
     5881 gaaagtatag gaact
//
