LOCUS       sequence-325433-        6203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Introduce Dox-inducible (TREa promoter) C124S-mutant Pten 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 6203)
  TITLE     Karreth lab plasmids
REFERENCE   2  (bases 1 to 6203)
  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-PtenC124S
FEATURES             Location/Qualifiers
     source          1..6203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..52
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1331..1903
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     rep_origin      complement(2231..2819)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2311..2330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2990..3781,3782..3850))
                     /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     3613..3632
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3851..3955)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3981..4436)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4118..4139)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4330..4349
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4563..4585
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4577..4594
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4578..4594
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4601..4620
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4601..4619
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4652..4668
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4652..4668
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4747..4770)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5109..5128
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    5222..5269
                     /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)."
     misc_feature    join(5302..5367,5368..5405)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     polyA_signal    5715..5849
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5752..5771)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5806..5825
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    join(5893..5911,5912..5934,5935..5953,5954..5976,5977..5995,
                     5996..6018,6019..6037,6038..6060,6061..6079,6080..6102,
                     6103..6121,6122..6144,6145..6163)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        join(6196..6203,1..31)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 tgtacggtgg gaggcctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag
       61 acgccatcca cgctcgaatg acagccatca tcaaagagat cgttagcaga aacaaaagga
      121 gatatcaaga ggatggattc gacttagact tgacctatat ttatccaaat attattgcta
      181 tgggatttcc tgcagaaaga cttgaaggtg tatacaggaa caatattgat gatgtagtaa
      241 ggtttttgga ttcaaagcat aaaaaccatt acaagatata caatctatgt gctgagagac
      301 attatgacac cgccaaattt aactgcagag ttgcacagta tccttttgaa gaccataacc
      361 caccacagct agaacttatc aaacccttct gtgaagatct tgaccaatgg ctaagtgaag
      421 atgacaatca tgttgcagca attcacagta aagctggaaa gggacggact ggtgtaatga
      481 tttgtgcata tttattgcat cggggcaaat ttttaaaggc acaagaggcc ctagattttt
      541 atggggaagt aaggaccaga gacaaaaagg gagtcacaat tcccagtcag aggcgctatg
      601 tatattatta tagctacctg ctaaaaaatc acctggatta cagacccgtg gcactgctgt
      661 ttcacaagat gatgtttgaa actattccaa tgttcagtgg cggaacttgc aatcctcagt
      721 ttgtggtctg ccagctaaag gtgaagatat attcctccaa ttcaggaccc acgcggcggg
      781 aggacaagtt catgtacttt gagttccctc agccattgcc tgtgtgtggt gatatcaaag
      841 tagagttctt ccacaaacag aacaagatgc tcaaaaagga caaaatgttt cacttttggg
      901 taaatacgtt cttcatacca ggaccagagg aaacctcaga aaaagtggaa aatggaagtc
      961 tttgtgatca ggaaatcgat agcatttgca gtatagagcg tgcagataat gacaaggagt
     1021 atcttgtact caccctaaca aaaaacgatc ttgacaaagc aaacaaagac aaggccaacc
     1081 gatacttctc tccaaatttt aaggtgaaac tatactttac aaaaacagta gaggagccat
     1141 caaatccaga ggctagcagt tcaacttctg tgactccaga tgttagtgac aatgaacctg
     1201 atcattatag atattctgac accactgact ctgatccaga gaatgaacct tttgatgaag
     1261 atcagcattc acaaattaca aaagtctaat cgagttcatg aattcgcagt cgagcgttga
     1321 gaacttcagg gtgagtttgg ggacccttga ttgttctttc tttttcgcta ttgtaaaatt
     1381 catgttatat ggagggggca aagttttcag ggtgttgttt agaatgggaa gatgtccctt
     1441 gtatcaccat ggaccctcat gataattttg tttctttcac tttctactct gttgacaacc
     1501 attgtctcct cttattttct tttcattttc tgtaactttt tcgttaaact ttagcttgca
     1561 tttgtaacga atttttaaat tcacttttgt ttatttgtca gattgtaagt actttctcta
     1621 atcacttttt tttcaaggca atcagggtat attatattgt acttcagcac agttttagag
     1681 aacaattgtt ataattaaat gataaggtag aatatttctg catataaatt ctggctggcg
     1741 tggaaatatt cttattggta gaaacaacta caccctggtc atcatcctgc ctttctcttt
     1801 atggttacaa tgatatacac tgtttgagat gaggataaaa tactctgagt ccaaaccggg
     1861 cccctctgct aaccatgttc atgccttctt ctctttccta cagctcctgg gcaacgtgct
     1921 ggttgttgtg ctgtctcatc attttggcaa aggattcact cctcaggtgc aggctgccta
     1981 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatcgttt
     2041 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     2101 ataaaggaaa tttattttca ttgcaatagt gtgttggaat ttctcgatcg ctagcatcct
     2161 aggcgcgcca catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2221 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2281 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2341 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2401 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2461 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2521 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2581 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2641 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2701 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2761 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2821 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2881 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2941 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3001 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3061 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3121 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3181 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3241 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3301 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3361 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3421 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3481 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3541 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3601 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3661 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3721 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3781 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3841 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3901 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3961 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     4021 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4081 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4141 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4201 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4261 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4321 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4381 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     4441 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     4501 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     4561 ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg
     4621 cgaattggag ctccaccgcg gtggcggccg ctctagaact agtggatccc ccgggctgca
     4681 ggaattaatt cgtcgacctc gaaattctac cgggtagggg aggcgctttt cccaaggcag
     4741 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
     4801 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
     4861 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
     4921 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc
     4981 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
     5041 ttcgctttct gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg
     5101 aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc
     5161 atctccgggc ctttcgacct gcatccatct agatctcgat cgagcagctg aagcttatgg
     5221 agaagttact attccgaagt tcctattctc tagaaagtat aggaacttcg atccccctcg
     5281 atcccccggg ctgcagatct gtagggcgca gtagtccagg gtttccttga tgatgtcata
     5341 cttatcctgt cccttttttt tccacagctc gcggttgagg acaaactctt cgcggtcttt
     5401 ccagtgggga tcgacggtat cgataagctt gcagatctgc gactctagag gatctgcgac
     5461 tctagaggat cataatcagc cataccacat ttgtagaggt ttagcaattc acagtaaagc
     5521 tggaaaggga cggactggtg taatgatttg tgcatattta ttgcatcggg gcaaattttt
     5581 aaagatgtct ggatctgcga ctctagagga tcataatcag ccataccaca tttgtagagg
     5641 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     5701 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5761 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5821 tcatcaatgt atcttatcat gtctggatcg agtttaccat gcatcttatc atgtctggat
     5881 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
     5941 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
     6001 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     6061 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
     6121 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagct
     6181 cggtacccgg gtcgaggtag gcg
//
