LOCUS       sequence-7051-ex        4706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4706)
  AUTHORS   Wong ET, Kolman JL, Li YC, Mesner LD, Hillen W, Berens C, Wahl GM
  TITLE     Reproducible doxycycline-inducible transgene expression at specific 
            loci generated by Cre-recombinase mediated cassette exchange.
  JOURNAL   Nucleic Acids Res. 2005  . 33(17):e147.
  PUBMED    16204450
REFERENCE   2  (bases 1 to 4706)
  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: "Nucleic 
            Acids Res. 2005  . 33(17):e147."
COMMENT     Sequence Label: pL3-TRE-MCS-polyA-2L
FEATURES             Location/Qualifiers
     source          1..4706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     428..444
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    477..510
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    join(541..559,560..582,583..601,602..624,625..643,644..666,
                     667..685,686..708,709..727,728..750,751..769,770..792,
                     793..811)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        844..882
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     879..903
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1080..1652
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1784..1803)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1849..1904
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1903..1922)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    complement(2418..2451)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2485..2501)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2485..2501)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2498..2520)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2509..2525
                     /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)."
     promoter        complement(join(2533..2539,2540..2557,2558..2563))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2578..2599
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2716..2733)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2887..3475)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2967..2986)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3646..4437,4438..4506))
                     /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     4269..4288
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4507..4611)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4679..4697
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccgggggc
      421 gcgccttgta aaacgacggc cagtgaattc tcgatctaga gaaccccttc atcgagataa
      481 cttcgtataa agtctcctat acgaagttat atcgatgaac cccttcctcg agtttaccac
      541 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
      601 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
      661 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
      721 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
      781 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagctcg gtacccgggt
      841 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
      901 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
      961 agcctccgcg gccccgaatt cgagctcggt acccggggat cctctagtca gctgacgcgt
     1021 gctagcgcgg ccgcatcgat aagcttgtcg acgatatctc tagagctgag aacttcaggg
     1081 tgagtttggg gacccttgat tgttctttct ttttcgctat tgtaaaattc atgttatatg
     1141 gagggggcaa agttttcagg gtgttgttta gaatgggaag atgtcccttg tatcaccatg
     1201 gaccctcatg ataattttgt ttctttcact ttctactctg ttgacaacca ttgtctcctc
     1261 ttattttctt ttcattttct gtaacttttt cgttaaactt tagcttgcat ttgtaacgaa
     1321 tttttaaatt cacttttgtt tatttgtcag attgtaagta ctttctctaa tcactttttt
     1381 ttcaaggcaa tcagggtata ttatattgta cttcagcaca gttttagaga acaattgtta
     1441 taattaaatg ataaggtaga atatttctgc atataaattc tggctggcgt ggaaatattc
     1501 ttattggtag aaacaactac atcctggtaa tcatcctgcc tttctcttta tggttacaat
     1561 gatatacact gtttgagatg aggataaaat actctgagtc caaaccgggc ccctctgcta
     1621 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttgttgtgc
     1681 tgtctcatca ttttggcaaa gaattcactc ctcaggtgca ggctgcctat cagaaggtgg
     1741 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
     1801 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctgggtaa taaaggaaat
     1861 ttattttcat tgcaatagtg tgtgggaatt ttttgtgtct ctcactcgga aggacatatg
     1921 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     1981 ccatatgctg gctgccatga acaaaggtgg ctataaagag gtcatcagta tatgaaacag
     2041 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     2101 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     2161 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     2221 tgaactcgac tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2281 cgctcttccg ctacgtcacc acccccggct ccataccgac gatatgcgac ctggcgcgca
     2341 cgtttgcccg ggagatgggg gaggctaact gaagcttccc gggggtaccg aattcgtcga
     2401 cagatccggc cggccccata acttcgtata gcatacatta tacgaagtta tcccctgcag
     2461 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2521 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2581 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2641 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2701 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2761 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2821 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2881 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2941 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3001 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3061 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3121 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3181 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3241 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3301 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     3361 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3421 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3481 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3541 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3601 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3661 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3721 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3781 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3841 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3901 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3961 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4021 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4081 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4141 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4201 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4261 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4321 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4381 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4441 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4501 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4561 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4621 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4681 taggcgtatc acgaggccct ttcgtc
//
