LOCUS       sequence-325426-        6351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Introduce Dox-inducible (TRE promoter) wildtype 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 6351)
  TITLE     Karreth lab plasmids
REFERENCE   2  (bases 1 to 6351)
  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-PtenWT
FEATURES             Location/Qualifiers
     source          1..6351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..547
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(83..106)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     480..499
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    593..640
                     /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(673..738,739..776)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     polyA_signal    945..1079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1199..1333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    join(1377..1395,1396..1418,1419..1437,1438..1460,1461..1479,
                     1480..1502,1503..1521,1522..1544,1545..1563,1564..1586,
                     1587..1605,1606..1628,1629..1647)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        1680..1718
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1715..1739
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          3018..3590
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     rep_origin      complement(3918..4506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3998..4017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4677..5468,5469..5537))
                     /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     5300..5319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5538..5642)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5668..6123)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5805..5826)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6017..6036
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6250..6272
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6264..6281
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6265..6281
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6288..6307
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6288..6306
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     join(6339..6351,1..4)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6339..6351,1..4)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 gatcccccgg gctgcaggaa ttaattcgtc gacctcgaaa ttctaccggg taggggaggc
       61 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
      121 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
      181 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
      241 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
      301 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
      361 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
      421 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
      481 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
      541 cctttcgacc tgcatccatc tagatctcga tcgagcagct gaagcttatg gagaagttac
      601 tattccgaag ttcctattct ctagaaagta taggaacttc gatccccctc gatcccccgg
      661 gctgcagatc tgtagggcgc agtagtccag ggtttccttg atgatgtcat acttatcctg
      721 tccctttttt ttccacagct cgcggttgag gacaaactct tcgcggtctt tccagtgggg
      781 atcgacggta tcgataagct tgcagatctg cgactctaga ggatctgcga ctctagagga
      841 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
      901 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
      961 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1021 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct
     1081 gcgactctag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1141 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1201 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1261 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1321 tcatgtctgg atcgagttta ccatgcatct tatcatgtct ggatcgagtt taccactccc
     1381 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
     1441 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
     1501 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
     1561 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
     1621 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agctcggtac ccgggtcgag
     1681 gtaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg
     1741 cctggagacg ccatccacgc tcgaatgaca gccatcatca aagagatcgt tagcagaaac
     1801 aaaaggagat atcaagagga tggattcgac ttagacttga cctatattta tccaaatatt
     1861 attgctatgg gatttcctgc agaaagactt gaaggtgtat acaggaacaa tattgatgat
     1921 gtagtaaggt ttttggattc aaagcataaa aaccattaca agatatacaa tctatgtgct
     1981 gagagacatt atgacaccgc caaatttaac tgcagagttg cacagtatcc ttttgaagac
     2041 cataacccac cacagctaga acttatcaaa cccttctgtg aagatcttga ccaatggcta
     2101 agtgaagatg acaatcatgt tgcagcaatt cactgtaaag ctggaaaggg acggactggt
     2161 gtaatgattt gtgcatattt attgcatcgg ggcaaatttt taaaggcaca agaggcccta
     2221 gatttttatg gggaagtaag gaccagagac aaaaagggag tcacaattcc cagtcagagg
     2281 cgctatgtat attattatag ctacctgcta aaaaatcacc tggattacag acccgtggca
     2341 ctgctgtttc acaagatgat gtttgaaact attccaatgt tcagtggcgg aacttgcaat
     2401 cctcagtttg tggtctgcca gctaaaggtg aagatatatt cctccaattc aggacccacg
     2461 cggcgggagg acaagttcat gtactttgag ttccctcagc cattgcctgt gtgtggtgat
     2521 atcaaagtag agttcttcca caaacagaac aagatgctca aaaaggacaa aatgtttcac
     2581 ttttgggtaa atacgttctt cataccagga ccagaggaaa cctcagaaaa agtggaaaat
     2641 ggaagtcttt gtgatcagga aatcgatagc atttgcagta tagagcgtgc agataatgac
     2701 aaggagtatc ttgtactcac cctaacaaaa aacgatcttg acaaagcaaa caaagacaag
     2761 gccaaccgat acttctctcc aaattttaag gtgaaactat actttacaaa aacagtagag
     2821 gagccatcaa atccagaggc tagcagttca acttctgtga ctccagatgt tagtgacaat
     2881 gaacctgatc attatagata ttctgacacc actgactctg atccagagaa tgaacctttt
     2941 gatgaagatc agcattcaca aattacaaaa gtctaatcga gttcatgaat tcgcagtcga
     3001 gcgttgagaa cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg
     3061 taaaattcat gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat
     3121 gtcccttgta tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt
     3181 gacaaccatt gtctcctctt attttctttt cattttctgt aactttttcg ttaaacttta
     3241 gcttgcattt gtaacgaatt tttaaattca cttttgttta tttgtcagat tgtaagtact
     3301 ttctctaatc actttttttt caaggcaatc agggtatatt atattgtact tcagcacagt
     3361 tttagagaac aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg
     3421 gctggcgtgg aaatattctt attggtagaa acaactacac cctggtcatc atcctgcctt
     3481 tctctttatg gttacaatga tatacactgt ttgagatgag gataaaatac tctgagtcca
     3541 aaccgggccc ctctgctaac catgttcatg ccttcttctc tttcctacag ctcctgggca
     3601 acgtgctggt tgttgtgctg tctcatcatt ttggcaaagg attcactcct caggtgcagg
     3661 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     3721 atcgttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     3781 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttc tcgatcgcta
     3841 gcatcctagg cgcgccacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3901 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3961 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4021 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4081 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4141 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4201 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4261 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4321 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4381 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4441 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4501 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4561 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4621 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4681 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4741 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4801 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4861 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4921 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4981 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5041 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5101 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5161 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5221 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5281 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5341 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5401 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5461 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5521 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5581 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5641 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     5701 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     5761 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     5821 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     5881 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     5941 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     6001 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     6061 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     6121 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     6181 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     6241 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac
     6301 tatagggcga attggagctc caccgcggtg gcggccgctc tagaactagt g
//
