LOCUS Exported 4549 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CreERT2 ORF with stop codon between attL1-attL2 on Gateway entry vector. ACCESSION . VERSION . KEYWORDS pENTR-CreERT2 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4549 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 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" 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 //