LOCUS       sequence-239592-        8104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac-based Dox-inducible expression of Esrrb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8104)
  AUTHORS   Adachi K, Kopp W, Wu G, Heising S, Greber B, Stehling M, 
            Arauzo-Bravo MJ, Boerno ST, Timmermann B, Vingron M, Scholer HR
  TITLE     Esrrb Unlocks Silenced Enhancers for Reprogramming to Naive 
            Pluripotency.
  JOURNAL   Cell Stem Cell. 2018 Aug 2;23(2):266-275.e6. doi: 
            10.1016/j.stem.2018.05.020. Epub 2018 Jun 14.
  PUBMED    29910149
REFERENCE   2  (bases 1 to 8104)
  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"; doi: 
            "10.1016/j.stem.2018.05.020"; journalName: "Cell Stem Cell"; date: 
            "2018-08-2- 2"; volume: "23"; issue: "2"; pages: "266-275.e6"
COMMENT     Sequence Label: KA1042_pPB-hCMV*1-Esrrb-IV
FEATURES             Location/Qualifiers
     source          1..8104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(186..204,205..227,228..246,247..269,270..288,289..311,
                     312..330,331..353,354..372,373..395,396..414,415..437,
                     438..456)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        489..527
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     524..548
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          650..1222
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     misc_feature    join(2632..3206,3207..3209,3210..3218)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2799..2816)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3026..3045
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3219..3221,3222..3224,3225..3938)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3264..3285)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3525..3544)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3872..3893
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4323..4340)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4329..4553
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4600..4616)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4624..4640
                     /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(4648..4654,4655..4672,4673..4678))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4693..4714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   4941..5003
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(5027..5045)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5066..5082)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5090..5106
                     /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(5114..5120,5121..5138,5139..5144))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5159..5180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5297..5314)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5468..6056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5548..5567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6227..7018,7019..7087))
                     /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     6850..6869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7088..7192)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7218..7673)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7355..7376)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7567..7586
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7800..7822
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7814..7831
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7815..7831
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7838..7857
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7838..7856
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   7875..7909
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 catacgataa ttatattgtt atttcatgtt ctacttacgt gataacttat tatatatata
       61 ttttcttgtt atagatatca taatttaaac aagcaaaacc aaattaaggg ccagctcatt
      121 cctcccactc atgatctata gatctggatc cgtcgacaac ggcaagctta catcgagttt
      181 accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga
      241 tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc
      301 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat
      361 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
      421 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gctcggtacc
      481 cgggtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc
      541 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
      601 gatccagcct ccgcgggact cagacacccg ggatcctgag aacttcaggg tgagtttggg
      661 gacccttgat tgttctttct ttttcgctat tgtaaaattc atgttatatg gagggggcaa
      721 agttttcagg gtgttgttta gaatgggaag atgtcccttg tatcaccatg gaccctcatg
      781 ataattttgt ttctttcact ttctactctg ttgacaacca ttgtctcctc ttattttctt
      841 ttcattttct gtaacttttt cgttaaactt tagcttgcat ttgtaacgaa tttttaaatt
      901 cacttttgtt tatttgtcag attgtaagta ctttctctaa tcactttttt ttcaaggcaa
      961 tcagggtata ttatattgta cttcagcaca gttttagaga acaattgtta taattaaatg
     1021 ataaggtaga atatttctgc atataaattc tggctggcgt ggaaatattc ttattggtag
     1081 aaacaactac accctggtca tcatcctgcc tttctcttta tggttacaat gatatacact
     1141 gtttgagatg aggataaaat actctgagtc caaaccgggc ccctctgcta accatgttca
     1201 tgccttcttc tctttcctac agctcctggg caacgtgctg gttgttgtgc tgtctcatca
     1261 ttttggcaaa gaattcctcg agaccaccat gtcgtccgaa gacaggcacc tgggctctag
     1321 ttgcggctcc ttcatcaaga cggagccatc cagcccgtcc tcgggcattg atgccctcag
     1381 ccaccacagc cccagcggct cgtcggacgc cagtggtggc tttggcattg ccctgagcac
     1441 ccacgccaac ggtctggact cgccgcctat gttcgcaggt gcggggctgg gaggcaaccc
     1501 gtgccgcaag agctacgagg actgtactag tggtatcatg gaggactccg ccatcaaatg
     1561 cgagtacatg cttaacgcca tccccaagcg cctgtgcctc gtgtgcgggg acattgcctc
     1621 tggctaccac tacggagtgg cctcctgcga ggcttgcaag gcgttcttca agagaaccat
     1681 tcaaggcaac atcgagtaca actgcccggc caccaatgaa tgtgagatca ccaaacggag
     1741 gcgcaagtcc tgtcaggcct gccgattcat gaaatgcctc aaagtgggga tgctgaagga
     1801 aggtgtgcgc cttgaccgag ttcgaggagg ccgccagaag tacaagcgac ggctggattc
     1861 ggagaacagc ccctacctga acctgccgat ttccccacct gctaaaaagc cattgactaa
     1921 gatcgtctcg aatctactag gggttgagca ggacaagctg tatgctatgc ctcccaacga
     1981 tatccccgag ggagatatca aggccctgac cactctctgt gaattggcag atcgggagct
     2041 tgtgttcctc atcaactggg ccaagcacat cccaggcttc cccagtctga cacttgggga
     2101 ccagatgagc ctgctgcaga gtgcctggat ggagattctc atcttgggca tcgtgtaccg
     2161 ctcgctccca tacgatgaca agctggcata cgccgaggac tatatcatgg atgaggaaca
     2221 ctctcgcctg gtagggctgc tggaccttta ccgagccatc ctgcagctgg tgcgcaggta
     2281 caagaaactc aaggtagaga aggaagagtt tatgatcctc aaggccctgg ccctcgccaa
     2341 ctcagattcg atgtacattg agaacctgga ggcggtgcag aagctccagg acctgctgca
     2401 cgaggcgctg caggactatg agctgagtca gcgccacgag gagccgcgga gggccggcaa
     2461 gctgctgctg acgctgcccc tgctgaggca gacagccgcc aaagccgtgc aacacttcta
     2521 cagtgtgaaa ctgcagggca aggtgcccat gcacaaactc ttcctggaga tgctggaggc
     2581 caaggtgtga tgcggccgct cgataagctt gatatcgaat tccgcccccc ccccctctcc
     2641 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     2701 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     2761 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     2821 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     2881 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     2941 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     3001 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     3061 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     3121 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     3181 tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag ggcgaggagc
     3241 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     3301 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga
     3361 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg
     3421 gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     3481 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     3541 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3601 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     3661 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
     3721 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
     3781 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc
     3841 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     3901 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccagctt atcgactcgc
     3961 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagt cgagaaattg
     4021 atgatctatt aaacaataaa gatgtccaca tggaagtttt tcctgtcata ctttgttaag
     4081 aagggtgaga acagagtacc tacattttga atggaaggat tggagctacg ggggtggggg
     4141 tggggtggga ttagataaat gcctgctctt tactgaaggc tctttactat tgctttatga
     4201 taatgtttca tagttggata tcataattta aacaagcaaa accaaattaa gggccagctc
     4261 attcctccca ctcatgatct atagatccta agcttatcga tgatctagag ctcgctgatc
     4321 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4381 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4441 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4501 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4561 ggcggaaaga acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4621 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4681 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4741 ccagtcggga aacctgtcgt gccagcggat cctttgttac tttatagaag aaattttgag
     4801 tttttgtttt tttttaataa ataaataaac ataaataaat tgtttgttga atttattatt
     4861 agtatgtaag tgtaaatata ataaaactta atatctattc aaattaataa ataaacctcg
     4921 atatacagac cgataaaaca catgcgtcaa ttttacgcat gattatcttt aacgtacgtc
     4981 acaatatgat tatctttcta gggttaaaga gctccagctt ttgttccctt tagtgagggt
     5041 taatttcgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5101 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5161 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5221 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5281 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5341 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5401 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5461 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5521 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5581 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5641 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5701 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5761 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5821 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5881 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5941 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6001 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6061 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6121 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6181 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6241 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6301 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6361 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6421 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6481 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6541 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6601 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6661 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6721 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6781 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6841 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6901 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6961 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7021 caaaaacggg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7081 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7141 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7201 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     7261 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     7321 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     7381 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     7441 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     7501 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     7561 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     7621 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     7681 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     7741 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     7801 ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga
     7861 attgggtacc ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa
     7921 tattgctctc tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc
     7981 gccgcttgga gctcccgtga ggcgtgcttg tcaatgaggt aagtgtcact gattttgaac
     8041 tataacgacc gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt
     8101 aact
//
