LOCUS Exported 5599 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION N-terminal part of the split-CreERT2 System. ACCESSION . VERSION . KEYWORDS pCMV-Tag2B-NCreERT2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5599) AUTHORS Hirrlinger J, Requardt RP, Winkler U, Wilhelm F, Schulze C, Hirrlinger PG TITLE Split-CreERT2: temporal control of DNA recombination mediated by split-Cre protein fragment complementation. JOURNAL PLoS One. 2009 Dec 16;4(12):e8354. doi: 10.1371/journal.pone.0008354. PUBMED 20016782 REFERENCE 2 (bases 1 to 5599) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5599 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 67..370 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 371..574 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 525..545 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" promoter 620..638 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" regulatory 673..682 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 682..705 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 790..846 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 919..939 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1072..2007 /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" primer_bind complement(2028..2044) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2029..2045) /label=pBluescriptKS /note="For pBluescript vector" primer_bind complement(2100..2119) /label=T7 /note="T7 promoter, forward primer" promoter complement(2101..2119) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 2393..2514 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2430..2449) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2484..2503 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2521..2976) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2658..2679) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2870..2889 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3003..3107 /gene="bla" /label=AmpR promoter promoter 3109..3466 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3132..3152) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3317..3452 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3379..3398 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3501..4295 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3555..3574) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4165..4184 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4483..4502) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4527..4574 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4903..5491 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5392..5411 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta 601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc 661 gcggtggcgg ccgccaccat ggattacaag gatgacgacg ataagagccc gggcggatcc 721 aacactgaag ccgccaggcg ttctcgtgcg agaaagttgc aaagaatgaa acaacttgaa 781 gacaaggttg aagaattgct ttcgaaaaat tatcacttgg aaaatgaggt tgccagatta 841 aagaaattag ttggcgaacg cgccagccct agcaaccctg gcgccagcaa cggcagcctg 901 caggaattca gcggccctcc caagaagaag aggaaggtga cctctgatga agtcaggaag 961 aacctgatgg acatgttcag ggacaggcag gccttctctg aacacacctg gaagatgctc 1021 ctgtctgtgt gcagatcctg ggctgcctgg tgcaagctga acactagttc tgctggagac 1081 atgagagctg ccaacctttg gccaagcccg ctcatgatca aacgctctaa gaagaacagc 1141 ctggccttgt ccctgacggc cgaccagatg gtcagtgcct tgttggatgc tgagcccccc 1201 atactctatt ccgagtatga tcctaccaga cccttcagtg aagcttcgat gatgggctta 1261 ctgaccaacc tggcagacag ggagctggtt cacatgatca actgggcgaa gagggtgcca 1321 ggctttgtgg atttgaccct ccatgatcag gtccaccttc tagaatgtgc ctggctagag 1381 atcctgatga ttggtctcgt ctggcgctcc atggagcacc cagtgaagct actgtttgct 1441 cctaacttgc tcttggacag gaaccaggga aaatgtgtag agggcatggt ggagatcttc 1501 gacatgctgc tggctacatc atctcggttc cgcatgatga atctgcaggg agaggagttt 1561 gtgtgcctca aatctattat tttgcttaat tctggagtgt acacatttct gtccagcacc 1621 ctgaagtctc tggaagagaa ggaccatatc caccgagtcc tggacaagat cacagacact 1681 ttgatccacc tgatggccaa ggcaggcctg accctgcagc agcagcacca gcggctggcc 1741 cagctcctcc tcatcctctc ccacatcagg cacatgagta acaaaggcat ggagcatctg 1801 tacagcatga agtgcaagaa cgtggtgccc ctctatgacc tgctgctgga ggcggcggac 1861 gcccaccgcc tacatgcgcc cactagccgt ggaggggcat ccgtggagga gacggaccaa 1921 agccacttgg ccactgcggg ctctacttca tcgcattcct tgcaaaagta ttacatcacg 1981 ggggaggcag agggtttccc tgccacagtc tgaactagta agcttatcga taccgtcgac 2041 ctcgaggggg ggcccggtac cttaattaat taaggtacca ggtaagtgta cccaattcgc 2101 cctatagtga gtcgtattac aattcactcg atcgcccttc ccaacagttg cgcagcctga 2161 atggcgaatg gagatccaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt 2221 ttgtgtattt tagattcaca gtcccaaggc tcatttcagg cccctcagtc ctcacagtct 2281 gttcatgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct 2341 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt 2401 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 2461 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt 2521 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat 2581 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga 2641 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca 2701 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct 2761 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc 2821 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag 2881 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca 2941 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg 3001 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga 3061 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagaatcct gaggcggaaa 3121 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 3181 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 3241 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 3301 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 3361 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt 3421 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga 3481 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg 3541 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 3601 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt 3661 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg 3721 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 3781 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 3841 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 3901 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 3961 atcaggatga tctggacgaa gaacatcagg ggctcgcgcc agccgaactg ttcgccaggc 4021 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc 4081 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 4141 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gaacttggcg 4201 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 4261 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac 4321 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga 4381 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga 4441 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac 4501 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt 4561 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc 4621 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc 4681 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc 4741 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat 4801 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt 4861 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 4921 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 4981 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 5041 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 5101 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 5161 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 5221 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 5281 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 5341 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 5401 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 5461 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 5521 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt 5581 ggataaccgt attaccgcc //