LOCUS       sequence-274672-        4549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CreERT2 ORF with stop codon between attL1-attL2 on Gateway entry 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4549)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 4549)
  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: pENTR-CreERT2
FEATURES             Location/Qualifiers
     source          1..4549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             680..1708
                     /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             1721..2656
                     /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             1769..2662
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     protein_bind    complement(2674..2773)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2790..2809)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2791..2809)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2814..2830)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2814..2830)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2932..2951)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2943..3752
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3010..3029)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3899..4487
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4388..4407
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctct cgagccacca tgtccaattt actgaccgta caccaaaatt tgcctgcatt
      721 accggtcgat gcaacgagtg atgaggttcg caagaacctg atggacatgt tcagggatcg
      781 ccaggcgttt tctgagcata cctggaaaat gcttctgtcc gtttgccggt cgtgggcggc
      841 atggtgcaag ttgaataacc ggaaatggtt tcccgcagaa cctgaagatg ttcgcgatta
      901 tcttctatat cttcaggcgc gcggtctggc agtaaaaact atccagcaac atttgggcca
      961 gctaaacatg cttcatcgtc ggtccgggct gccacgacca agtgacagca atgctgtttc
     1021 actggttatg cggcggatcc gaaaagaaaa cgttgatgcc ggtgaacgtg caaaacaggc
     1081 tctagcgttc gaacgcactg atttcgacca ggttcgttca ctcatggaaa atagcgatcg
     1141 ctgccaggat atacgtaatc tggcatttct ggggattgct tataacaccc tgttacgtat
     1201 agccgaaatt gccaggatca gggttaaaga tatctcacgt actgacggtg ggagaatgtt
     1261 aatccatatt ggcagaacga aaacgctggt tagcaccgca ggtgtagaga aggcacttag
     1321 cctgggggta actaaactgg tcgagcgatg gatttccgtc tctggtgtag ctgatgatcc
     1381 gaataactac ctgttttgcc gggtcagaaa aaatggtgtt gccgcgccat ctgccaccag
     1441 ccagctatca actcgcgccc tggaagggat ttttgaagca actcatcgat tgatttacgg
     1501 cgctaaggat gactctggtc agagatacct ggcctggtct ggacacagtg cccgtgtcgg
     1561 agccgcgcga gatatggccc gcgctggagt ttcaataccg gagatcatgc aagctggtgg
     1621 ctggaccaat gtaaatattg tcatgaacta tatccgtaac ctggatagtg aaacaggggc
     1681 aatggtgcgc ctgctggaag atggcgatct cgagccatct gctggagaca tgagagctgc
     1741 caacctttgg ccaagcccgc tcatgatcaa acgctctaag aagaacagcc tggccttgtc
     1801 cctgacggcc gaccagatgg tcagtgcctt gttggatgct gagcccccca tactctattc
     1861 cgagtatgat cctaccagac ccttcagtga agcttcgatg atgggcttac tgaccaacct
     1921 ggcagacagg gagctggttc acatgatcaa ctgggcgaag agggtgccag gctttgtgga
     1981 tttgaccctc catgatcagg tccaccttct agaatgtgcc tggctagaga tcctgatgat
     2041 tggtctcgtc tggcgctcca tggagcaccc agtgaagcta ctgtttgctc ctaacttgct
     2101 cttggacagg aaccagggaa aatgtgtaga gggcatggtg gagatcttcg acatgctgct
     2161 ggctacatca tctcggttcc gcatgatgaa tctgcaggga gaggagtttg tgtgcctcaa
     2221 atctattatt ttgcttaatt ctggagtgta cacatttctg tccagcaccc tgaagtctct
     2281 ggaagagaag gaccatatcc accgagtcct ggacaagatc acagacactt tgatccacct
     2341 gatggccaag gcaggcctga ccctgcagca gcagcaccag cggctggccc agctcctcct
     2401 catcctctcc cacatcaggc acatgagtaa caaaggcatg gagcatctgt acagcatgaa
     2461 gtgcaagaac gtggtgcccc tctatgacct gctgctggag gcggcggacg cccaccgcct
     2521 acatgcgccc actagccgtg gaggggcatc cgtggaggag acggaccaaa gccacttggc
     2581 cactgcgggc tctacttcat cgcattcctt gcaaaagtat tacatcacgg gggaggcaga
     2641 gggtttccct gccacagctt gataggcggc cgcacccagc tttcttgtac aaagttggca
     2701 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     2761 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     2821 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     2881 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     2941 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     3001 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     3061 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     3121 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     3181 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     3241 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     3301 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     3361 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     3421 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     3481 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     3541 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     3601 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3661 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3721 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     3781 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     3841 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3901 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3961 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4021 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4081 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4141 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4201 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4261 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4321 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4381 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4441 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4501 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
