LOCUS       sequence-154036-        8135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG promoter driving Cre-ERT2 fusion-IRES-Blasticidin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8135)
  AUTHORS   Betschinger J, Nichols J, Dietmann S, Corrin PD, Paddison PJ, Smith 
            A
  TITLE     Exit from pluripotency is gated by intracellular redistribution of 
            the bHLH transcription factor Tfe3.
  JOURNAL   Cell. 2013 Apr 11;153(2):335-47. doi: 10.1016/j.cell.2013.03.012.
  PUBMED    23582324
REFERENCE   2  (bases 1 to 8135)
  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.cell.2013.03"; journalName: "Cell"; date: "2013-04-11- 
            11"; volume: "153"; issue: "2"; pages: "335-47"
COMMENT     Sequence Label: pPy-CAG-Cre::ERT2-IRES-BSD
FEATURES             Location/Qualifiers
     source          1..8135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        380..655
                     /label=chicken beta-actin promoter
     intron          656..1664
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1588..1610
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1725..2753
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             2766..3701
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             2814..3707
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     misc_feature    join(3741..4314,4315..4317,4318..4326)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3908..3925)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4135..4154
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(4739..4756)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4745..4969
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5016..5032)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5016..5032)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5029..5051)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5040..5056
                     /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(5064..5070,5071..5088,5089..5094))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5109..5130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5189..5384
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5235..5370
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5390..5524
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5592..5609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5763..6351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5843..5862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6522..7313,7314..7382))
                     /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     7145..7164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    7985..8119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        join(8133..8135,1..377)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
       61 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      121 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      181 tccattgacg tcaatgggtg gactatttac ggtaaactgc ccacttggca gtacatcaag
      241 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      301 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      361 tcatcgctat taccatgggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
      421 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
      481 ggggcggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg
      541 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
      601 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
      661 cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc cgccccggct
      721 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
      781 taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg aaagccttaa
      841 agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg tgcgtgtgtg
      901 tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc gctgcgggcg
      961 cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg gggcggtgcc
     1021 ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     1081 ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca cccccctccc
     1141 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtgcggggcg tggcgcgggg
     1201 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
     1261 cgggccgggg agggctcggg ggaggggcgc ggcggccccg gagcgccggc ggctgtcgag
     1321 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     1381 ctttgtccca aatctggcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg
     1441 gcgcgggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
     1501 tcgccgcgcc gccgtcccct tctccatctc cagcctcggg gctgccgcag ggggacggct
     1561 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     1621 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     1681 tggttgttgt gctgtctcat cattttggca aagaattcct cgacatgtcc aatttactga
     1741 ccgtacacca aaatttgcct gcattaccgg tcgatgcaac gagtgatgag gttcgcaaga
     1801 acctgatgga catgttcagg gatcgccagg cgttttctga gcatacctgg aaaatgcttc
     1861 tgtccgtttg ccggtcgtgg gcggcatggt gcaagttgaa taaccggaaa tggtttcccg
     1921 cagaacctga agatgttcgc gattatcttc tatatcttca ggcgcgcggt ctggcagtaa
     1981 aaactatcca gcaacatttg ggccagctaa acatgcttca tcgtcggtcc gggctgccac
     2041 gaccaagtga cagcaatgct gtttcactgg ttatgcggcg gatccgaaaa gaaaacgttg
     2101 atgccggtga acgtgcaaaa caggctctag cgttcgaacg cactgatttc gaccaggttc
     2161 gttcactcat ggaaaatagc gatcgctgcc aggatatacg taatctggca tttctgggga
     2221 ttgcttataa caccctgtta cgtatagccg aaattgccag gatcagggtt aaagatatct
     2281 cacgtactga cggtgggaga atgttaatcc atattggcag aacgaaaacg ctggttagca
     2341 ccgcaggtgt agagaaggca cttagcctgg gggtaactaa actggtcgag cgatggattt
     2401 ccgtctctgg tgtagctgat gatccgaata actacctgtt ttgccgggtc agaaaaaatg
     2461 gtgttgccgc gccatctgcc accagccagc tatcaactcg cgccctggaa gggatttttg
     2521 aagcaactca tcgattgatt tacggcgcta aggatgactc tggtcagaga tacctggcct
     2581 ggtctggaca cagtgcccgt gtcggagccg cgcgagatat ggcccgcgct ggagtttcaa
     2641 taccggagat catgcaagct ggtggctgga ccaatgtaaa tattgtcatg aactatatcc
     2701 gtaacctgga tagtgaaaca ggggcaatgg tgcgcctgct ggaagatggc gatctcgagc
     2761 catctgctgg agacatgaga gctgccaacc tttggccaag cccgctcatg atcaaacgct
     2821 ctaagaagaa cagcctggcc ttgtccctga cggccgacca gatggtcagt gccttgttgg
     2881 atgctgagcc ccccatactc tattccgagt atgatcctac cagacccttc agtgaagctt
     2941 cgatgatggg cttactgacc aacctggcag acagggagct ggttcacatg atcaactggg
     3001 cgaagagggt gccaggcttt gtggatttga ccctccatga tcaggtccac cttctagaat
     3061 gtgcctggct agagatcctg atgattggtc tcgtctggcg ctccatggag cacccagtga
     3121 agctactgtt tgctcctaac ttgctcttgg acaggaacca gggaaaatgt gtagagggca
     3181 tggtggagat cttcgacatg ctgctggcta catcatctcg gttccgcatg atgaatctgc
     3241 agggagagga gtttgtgtgc ctcaaatcta ttattttgct taattctgga gtgtacacat
     3301 ttctgtccag caccctgaag tctctggaag agaaggacca tatccaccga gtcctggaca
     3361 agatcacaga cactttgatc cacctgatgg ccaaggcagg cctgaccctg cagcagcagc
     3421 accagcggct ggcccagctc ctcctcatcc tctcccacat caggcacatg agtaacaaag
     3481 gcatggagca tctgtacagc atgaagtgca agaacgtggt gcccctctat gacctgctgc
     3541 tggaggcggc ggacgcccac cgcctacatg cgcccactag ccgtggaggg gcatccgtgg
     3601 aggagacgga ccaaagccac ttggccactg cgggctctac ttcatcgcat tccttgcaaa
     3661 agtattacat cacgggggag gcagagggtt tccctgccac agtctgagcg gccgctcgat
     3721 aagcttgata tcgaattccg cccctctccc tccccccccc ctaacgttac tggccgaagc
     3781 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     3841 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     3901 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     3961 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     4021 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     4081 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     4141 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     4201 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaacgtc
     4261 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
     4321 acaaccatgc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
     4381 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
     4441 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
     4501 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
     4561 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
     4621 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
     4681 cgtgaattgc tgccctctgg ttatgtgtgg gagggttaac tagataagta atgatcagcc
     4741 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     4801 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4861 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4921 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4981 gaaagaacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5041 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5101 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5161 tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc
     5221 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5281 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5341 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat
     5401 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5461 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5521 gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5581 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5641 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5701 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5761 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5821 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5881 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5941 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6001 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6061 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6121 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6181 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6241 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6301 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6361 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6421 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6481 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6541 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6601 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6661 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6721 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6781 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6841 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6901 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6961 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7021 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7081 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7141 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7201 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7261 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7321 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7381 atactcttcc tttttcaatg accgctttgg aaaaacaaag actgtatttc ctggaaatta
     7441 atgtttattc aataaactgt gtattcagct atatcacata gtggtgaggc tgaaatgagg
     7501 cgggaagagg cggttggggc ttaattatat caatttgggt ggccccacag cgcctccaag
     7561 gcgccagtcc tgttttgaca agttgcctct ggaagcctct acaatgcctc tcttcttttt
     7621 ctccagagta agcggaggcc aggggccccc ggcctctgct taatactaaa aaaaacagct
     7681 gttgtcatag taatgattgg gtggaaacat tccaggcctg ggtggagagg ctttttgctt
     7741 cctcttgcaa aaccacactg acattccagg cctgggtgga gaggcttttt gcttcctctt
     7801 gcaaaaccac actgccctct ggagggcagt tgcctagcaa ctaactaaaa gaggatgtcg
     7861 cacggccagc tgcggtcagt tagtcacttc ctgcttaact gacttgacat tttctatttt
     7921 aagagtcggg aggaaaatta ctgtgttgga ggccctccgc catcttctga agctgaatcg
     7981 aattaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     8041 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     8101 atcttatcat gtctggatct gatatcatcg tcgac
//
