LOCUS       sequence-261849-        6344 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 6344)
  TITLE     Karreth lab plasmids
REFERENCE   2  (bases 1 to 6344)
  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..6344
                     /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(663..728,729..766)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     polyA_signal    935..1069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1189..1323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    join(1367..1385,1386..1408,1409..1427,1428..1450,1451..1469,
                     1470..1492,1493..1511,1512..1534,1535..1553,1554..1576,
                     1577..1595,1596..1618,1619..1637)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        1670..1708
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1705..1729
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          3008..3580
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     rep_origin      complement(3908..4496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3988..4007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4667..5458,5459..5527))
                     /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     5290..5309
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5528..5632)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5658..6113)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5795..5816)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6007..6026
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6240..6262
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6254..6271
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6255..6271
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6281..6300
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6281..6299
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     join(6332..6344,1..4)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6332..6344,1..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 gatcccccgg gctgcagatc
      661 tgtagggcgc agtagtccag ggtttccttg atgatgtcat acttatcctg tccctttttt
      721 ttccacagct cgcggttgag gacaaactct tcgcggtctt tccagtgggg atcgacggta
      781 tcgataagct tgcagatctg cgactctaga ggatctgcga ctctagagga tcataatcag
      841 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
      901 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
      961 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     1021 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct gcgactctag
     1081 aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     1141 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     1201 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1261 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     1321 atcgagttta ccatgcatct tatcatgtct ggatcgagtt taccactccc tatcagtgat
     1381 agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg
     1441 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc
     1501 agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga
     1561 aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc
     1621 cctatcagtg atagagaaaa gtgaaagtcg agctcggtac ccgggtcgag gtaggcgtgt
     1681 acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
     1741 ccatccacgc tcgaatgaca gccatcatca aagagatcgt tagcagaaac aaaaggagat
     1801 atcaagagga tggattcgac ttagacttga cctatattta tccaaatatt attgctatgg
     1861 gatttcctgc agaaagactt gaaggtgtat acaggaacaa tattgatgat gtagtaaggt
     1921 ttttggattc aaagcataaa aaccattaca agatatacaa tctatgtgct gagagacatt
     1981 atgacaccgc caaatttaac tgcagagttg cacagtatcc ttttgaagac cataacccac
     2041 cacagctaga acttatcaaa cccttctgtg aagatcttga ccaatggcta agtgaagatg
     2101 acaatcatgt tgcagcaatt cactgtaaag ctggaaaggg acggactggt gtaatgattt
     2161 gtgcatattt attgcatcgg ggcaaatttt taaaggcaca agaggcccta gatttttatg
     2221 gggaagtaag gaccagagac aaaaagggag tcacaattcc cagtcagagg cgctatgtat
     2281 attattatag ctacctgcta aaaaatcacc tggattacag acccgtggca ctgctgtttc
     2341 acaagatgat gtttgaaact attccaatgt tcagtggcgg aacttgcaat cctcagtttg
     2401 tggtctgcca gctaaaggtg aagatatatt cctccaattc aggacccacg cggcgggagg
     2461 acaagttcat gtactttgag ttccctcagc cattgcctgt gtgtggtgat atcaaagtag
     2521 agttcttcca caaacagaac aagatgctca aaaaggacaa aatgtttcac ttttgggtaa
     2581 atacgttctt cataccagga ccagaggaaa cctcagaaaa agtggaaaat ggaagtcttt
     2641 gtgatcagga aatcgatagc atttgcagta tagagcgtgc agataatgac aaggagtatc
     2701 ttgtactcac cctaacaaaa aacgatcttg acaaagcaaa caaagacaag gccaaccgat
     2761 acttctctcc aaattttaag gtgaaactat actttacaaa aacagtagag gagccatcaa
     2821 atccagaggc tagcagttca acttctgtga ctccagatgt tagtgacaat gaacctgatc
     2881 attatagata ttctgacacc actgactctg atccagagaa tgaacctttt gatgaagatc
     2941 agcattcaca aattacaaaa gtctaatcga gttcatgaat tcgcagtcga gcgttgagaa
     3001 cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg taaaattcat
     3061 gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat gtcccttgta
     3121 tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt gacaaccatt
     3181 gtctcctctt attttctttt cattttctgt aactttttcg ttaaacttta gcttgcattt
     3241 gtaacgaatt tttaaattca cttttgttta tttgtcagat tgtaagtact ttctctaatc
     3301 actttttttt caaggcaatc agggtatatt atattgtact tcagcacagt tttagagaac
     3361 aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg gctggcgtgg
     3421 aaatattctt attggtagaa acaactacac cctggtcatc atcctgcctt tctctttatg
     3481 gttacaatga tatacactgt ttgagatgag gataaaatac tctgagtcca aaccgggccc
     3541 ctctgctaac catgttcatg ccttcttctc tttcctacag ctcctgggca acgtgctggt
     3601 tgttgtgctg tctcatcatt ttggcaaagg attcactcct caggtgcagg ctgcctatca
     3661 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atcgttttcc
     3721 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     3781 aaggaaattt attttcattg caatagtgtg ttggaatttc tcgatcgcta gcatcctagg
     3841 cgcgccacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3901 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3961 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4021 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4081 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4141 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4201 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4261 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4321 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4381 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4441 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4501 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4561 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4621 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4681 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4741 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4801 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4861 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4921 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4981 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5041 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5101 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5161 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5221 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5281 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5341 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5401 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5461 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5521 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5581 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5641 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     5701 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     5761 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     5821 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     5881 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     5941 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     6001 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     6061 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     6121 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6181 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6241 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
     6301 cgaattggag ctccaccgcg gtggcggccg ctctagaact agtg
//
