LOCUS Exported 7493 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Monomeric TagBFP2 and iCre-ERT2 recombinase (PV N-terminus). ACCESSION . VERSION . KEYWORDS pCAGGS-mTagBFP2-T2A-iCreERT2 (PV) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7493) AUTHORS Nam HS, Capecchi MR TITLE 2A creER optimization plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 7493) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7493 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 382..659 /label=chicken beta-actin promoter intron 660..1676 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1600..1622 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1684..1703 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1734..2444 /codon_start=1 /product="enhanced monomeric blue fluorescent protein (Subach et al., 2011)" /label=mTagBFP2 /note="mammalian codon-optimized" /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV ARYCDLPSKLGHKLN" CDS 2466..2519 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 3558..4493 /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(4511..4530) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 4576..4631 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(4630..4649) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4992..5008) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4992..5008) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5005..5027) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5016..5032 /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(5040..5070) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5085..5106 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5165..5360 /label=SV40 promoter /note="SV40 early promoter" rep_origin 5211..5346 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5273..5292 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 5366..5500 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5403..5422) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5457..5476 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5568..5585) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5739..6327) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5819..5838) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6498..7358) /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 7121..7140 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7359..7463) /gene="bla" /label=AmpR promoter ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct 421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga 481 tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc ggggggcggg 541 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc 601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg 661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc 721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc 781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa 841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg 901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct 961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg 1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt 1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc 1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg 1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg 1261 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg 1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc 1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc 1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg 1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc 1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga 1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc 1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc gccatggtgt 1741 ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg 1801 tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc 1861 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg 1921 ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact 1981 tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg 2041 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg 2101 tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg 2161 gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg 2221 acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag accacatata 2281 gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac 2341 tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg 2401 ccagatactg cgacctccct agcaaactgg ggcacaagct taatggcggc tccggcggcc 2461 gctccgaggg ccgcggcagc ctgctgacat gcggcgacgt ggaagagaac cctggccctg 2521 tgaacctgct gactgtgcac caaaacctgc ctgccctccc tgtggatgcc acctctgatg 2581 aagtcaggaa gaacctgatg gacatgttca gggacaggca ggccttctct gaacacacct 2641 ggaagatgct cctgtctgtg tgcagatcct gggctgcctg gtgcaagctg aacaacagga 2701 aatggttccc tgctgaacct gaggatgtga gggactacct cctgtacctg caagccagag 2761 gcctggctgt gaagaccatc caacagcacc tgggccagct caacatgctg cacaggagat 2821 ctggcctgcc tcgcccttct gactccaatg ctgtgtccct ggtgatgagg agaatcagaa 2881 aggagaatgt ggatgctggg gagagagcca agcaggccct ggcctttgaa cgcactgact 2941 ttgaccaagt cagatccctg atggagaact ctgacagatg ccaggacatc aggaacctgg 3001 ccttcctggg cattgcctac aacaccctgc tgcgcattgc cgaaattgcc agaatcagag 3061 tgaaggacat ctcccgcacc gatggtggga gaatgctgat ccacattggc aggaccaaga 3121 ccctggtgtc cacagctggt gtggagaagg ccctgtccct gggggttacc aagctggtgg 3181 agagatggat ctctgtgtct ggtgtggctg atgaccccaa caactacctg ttctgccggg 3241 tcagaaagaa tggtgtggct gccccttctg ccacctccca actgtccacc cgggccctgg 3301 aagggatctt tgaggccacc caccgcctga tctatggtgc caaggatgac tctgggcaga 3361 gatacctggc ctggtctggc cactctgcca gagtgggtgc tgccagggac atggccaggg 3421 ctggtgtgtc catccctgaa atcatgcagg ctggtggctg gaccaatgtg aacattgtga 3481 tgaactacat cagaaacctg gactctgaga ctggggccat ggtgaggctg ctagaggatg 3541 gggacctcga gccatctgct ggagacatga gagctgccaa cctttggcca agcccgctca 3601 tgatcaaacg ctctaagaag aacagcctgg ccttgtccct gacggccgac cagatggtca 3661 gtgccttgtt ggatgctgag ccccccatac tctattccga gtatgatcct accagaccct 3721 tcagtgaagc ttcgatgatg ggcttactga ccaacctggc agacagggag ctggttcaca 3781 tgatcaactg ggcgaagagg gtgccaggct ttgtggattt gaccctccat gatcaggtcc 3841 accttctaga atgtgcctgg ctagagatcc tgatgattgg tctcgtctgg cgctccatgg 3901 agcacccagt gaagctactg tttgctccta acttgctctt ggacaggaac cagggaaaat 3961 gtgtagaggg catggtggag atcttcgaca tgctgctggc tacatcatct cggttccgca 4021 tgatgaatct gcagggagag gagtttgtgt gcctcaaatc tattattttg cttaattctg 4081 gagtgtacac atttctgtcc agcaccctga agtctctgga agagaaggac catatccacc 4141 gagtcctgga caagatcaca gacactttga tccacctgat ggccaaggca ggcctgaccc 4201 tgcagcagca gcaccagcgg ctggcccagc tcctcctcat cctctcccac atcaggcaca 4261 tgagtaacaa aggcatggag catctgtaca gcatgaagtg caagaacgtg gtgcccctct 4321 atgacctgct gctggaggcg gcggacgccc accgcctaca tgcgcccact agccgtggag 4381 gggcatccgt ggaggagacg gaccaaagcc acttggccac tgcgggctct acttcatcgc 4441 attccttgca aaagtattac atcacggggg aggcagaggg tttccctgcc acagcttgag 4501 gcgcgccgga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc 4561 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt 4621 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt 4681 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc 4741 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa 4801 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa 4861 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac 4921 tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg 4981 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 5041 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 5101 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg 5161 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 5221 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 5281 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 5341 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata 5401 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 5461 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc 5521 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 5581 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 5641 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 5701 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 5761 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 5821 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 5881 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 5941 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 6001 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 6061 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 6121 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 6181 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 6241 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 6301 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 6361 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 6421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 6481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 6541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 6601 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 6661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 6721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 6781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 6841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 6901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 6961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 7021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 7081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 7141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 7201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 7261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 7321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 7381 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 7441 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg gtc //