LOCUS Exported 4739 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pENTR/D_creERt2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4739) AUTHORS Mosimann C, Kaufman CK, Li P, Pugach EK, Tamplin OJ, Zon LI TITLE Ubiquitous transgene expression and Cre-based recombination driven by the ubiquitin promoter in zebrafish. JOURNAL Development. 2011 Jan . 138(1):169-77. PUBMED 21138979 REFERENCE 2 (bases 1 to 4739) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4739 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 27..615 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 516..535 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 769..786 /label=L4440 /note="L4440 vector, forward primer" terminator 945..972 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 1064..1150 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 1199..1221 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1213..1230 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1214..1230 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 1246..1345 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 1366..2394 /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 2407..3342 /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" polyA_signal 3394..3515 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3431..3450) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3485..3504 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" protein_bind complement(3541..3640) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(3657..3676) /label=T7 /note="T7 promoter, forward primer" promoter complement(3658..3676) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3681..3697) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3681..3697) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3799..3818) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 3810..4619 /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(3877..3896) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" ORIGIN 1 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 61 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 121 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 181 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 241 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 301 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 361 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 421 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 481 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 541 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 601 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 661 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 721 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 781 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt 841 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca 901 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca 961 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc 1021 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga 1081 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc 1141 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc 1201 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccccaaat aatgatttta 1261 ttttgactga tagtgacctg ttcgttgcaa caaattgatg agcaatgctt ttttataatg 1321 ccaactttgt acaaaaaagc aggctccgcg gccgccccct tcaccatgtc caatttactg 1381 accgtacacc aaaatttgcc tgcattaccg gtcgatgcaa cgagtgatga ggttcgcaag 1441 aacctgatgg acatgttcag ggatcgccag gcgttttctg agcatacctg gaaaatgctt 1501 ctgtccgttt gccggtcgtg ggcggcatgg tgcaagttga ataaccggaa atggtttccc 1561 gcagaacctg aagatgttcg cgattatctt ctatatcttc aggcgcgcgg tctggcagta 1621 aaaactatcc agcaacattt gggccagcta aacatgcttc atcgtcggtc cgggctgcca 1681 cgaccaagtg acagcaatgc tgtttcactg gttatgcggc ggatccgaaa agaaaacgtt 1741 gatgccggtg aacgtgcaaa acaggctcta gcgttcgaac gcactgattt cgaccaggtt 1801 cgttcactca tggaaaatag cgatcgctgc caggatatac gtaatctggc atttctgggg 1861 attgcttata acaccctgtt acgtatagcc gaaattgcca ggatcagggt taaagatatc 1921 tcacgtactg acggtgggag aatgttaatc catattggca gaacgaaaac gctggttagc 1981 accgcaggtg tagagaaggc acttagcctg ggggtaacta aactggtcga gcgatggatt 2041 tccgtctctg gtgtagctga tgatccgaat aactacctgt tttgccgggt cagaaaaaat 2101 ggtgttgccg cgccatctgc caccagccag ctatcaactc gcgccctgga agggattttt 2161 gaagcaactc atcgattgat ttacggcgct aaggatgact ctggtcagag atacctggcc 2221 tggtctggac acagtgcccg tgtcggagcc gcgcgagata tggcccgcgc tggagtttca 2281 ataccggaga tcatgcaagc tggtggctgg accaatgtaa atattgtcat gaactatatc 2341 cgtaacctgg atagtgaaac aggggcaatg gtgcgcctgc tggaagatgg cgatctcgag 2401 ccatctgctg gagacatgag agctgccaac ctttggccaa gcccgctcat gatcaaacgc 2461 tctaagaaga acagcctggc cttgtccctg acggccgacc agatggtcag tgccttgttg 2521 gatgctgagc cccccatact ctattccgag tatgatccta ccagaccctt cagtgaagct 2581 tcgatgatgg gcttactgac caacctggca gacagggagc tggttcacat gatcaactgg 2641 gcgaagaggg tgccaggctt tgtggatttg accctccatg atcaggtcca ccttctagaa 2701 tgtgcctggc tagagatcct gatgattggt ctcgtctggc gctccatgga gcacccagtg 2761 aagctactgt ttgctcctaa cttgctcttg gacaggaacc agggaaaatg tgtagagggc 2821 atggtggaga tcttcgacat gctgctggct acatcatctc ggttccgcat gatgaatctg 2881 cagggagagg agtttgtgtg cctcaaatct attattttgc ttaattctgg agtgtacaca 2941 tttctgtcca gcaccctgaa gtctctggaa gagaaggacc atatccaccg agtcctggac 3001 aagatcacag acactttgat ccacctgatg gccaaggcag gcctgaccct gcagcagcag 3061 caccagcggc tggcccagct cctcctcatc ctctcccaca tcaggcacat gagtaacaaa 3121 ggcatggagc atctgtacag catgaagtgc aagaacgtgg tgcccctcta tgacctgctg 3181 ctggaggcgg cggacgccca ccgcctacat gcgcccacta gccgtggagg ggcatccgtg 3241 gaggagacgg accaaagcca cttggccact gcgggctcta cttcatcgca ttccttgcaa 3301 aagtattaca tcacggggga ggcagagggt ttccctgcca cagcttgatg aagatctgag 3361 ctccctggcg gaattcggat cttattaaag cagaacttgt ttattgcagc ttataatggt 3421 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 3481 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgaagggt gggcgcgccg 3541 acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca 3601 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct 3661 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa 3721 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg 3781 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc 3841 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg 3901 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt 3961 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa 4021 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg 4081 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat 4141 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt 4201 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc 4261 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct 4321 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag 4381 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag 4441 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat 4501 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta 4561 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat 4621 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg 4681 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagac //