LOCUS Exported 7502 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Monomeric TagBFP2 and iCre-ERT2 recombinase (PAAAA N-terminus). ACCESSION . VERSION . KEYWORDS pCAGGS-mTagBFP2-T2A-iCreERT2 (PAAAA) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7502) AUTHORS Nam HS, Capecchi MR TITLE 2A creER optimization plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 7502) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7502 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 11..390 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 670..1686 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1610..1632 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1694..1713 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1744..2454 /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 2476..2529 /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 3577..4512 /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(4530..4549) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 4595..4650 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(4649..4668) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(5011..5027) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5011..5027) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5024..5046) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5035..5051 /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(5059..5089) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5104..5125 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5184..5379 /label=SV40 promoter /note="SV40 early promoter" rep_origin 5230..5365 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5292..5311 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 5385..5519 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5422..5441) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5476..5495 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5587..5604) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5758..6346) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5838..5857) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6517..7377) /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 7140..7159 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7378..7482) /gene="bla" /label=AmpR promoter ORIGIN 1 cacctgggtc gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta 61 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc 121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg 181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg 241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa 301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac 361 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact 421 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt 481 tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc cggggggggg gggggggggg 541 gggggggggg ggggggsgrg gsggagaggt gcggcggcag ccaatcagag cggcgcgctc 601 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg 661 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg 721 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc 781 ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg 841 ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg 901 ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc 961 tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc 1021 gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg 1081 cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc 1141 cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt 1201 acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg 1261 ggcggggcgg ggccgcctcg ggccggggrg ggctcggggg rggggcgcgg cggcccccgg 1321 agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc 1381 gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc 1441 cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg 1501 ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg 1561 ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc 1621 tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc 1681 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc 1741 gccatggtgt ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg 1801 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac 1861 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc 1921 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc 1981 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca 2041 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc 2101 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag 2161 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa 2221 ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag 2281 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg 2341 gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag 2401 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct taatggcggc 2461 tccggcggcc gctccgaggg ccgcggcagc ctgctgacat gcggcgacgt ggaagagaac 2521 cctggccctg ccgctgcagc caacctgctg actgtgcacc aaaacctgcc tgccctccct 2581 gtggatgcca cctctgatga agtcaggaag aacctgatgg acatgttcag ggacaggcag 2641 gccttctctg aacacacctg gaagatgctc ctgtctgtgt gcagatcctg ggctgcctgg 2701 tgcaagctga acaacaggaa atggttccct gctgaacctg aggatgtgag ggactacctc 2761 ctgtacctgc aagccagagg cctggctgtg aagaccatcc aacagcacct gggccagctc 2821 aacatgctgc acaggagatc tggcctgcct cgcccttctg actccaatgc tgtgtccctg 2881 gtgatgagga gaatcagaaa ggagaatgtg gatgctgggg agagagccaa gcaggccctg 2941 gcctttgaac gcactgactt tgaccaagtc agatccctga tggagaactc tgacagatgc 3001 caggacatca ggaacctggc cttcctgggc attgcctaca acaccctgct gcgcattgcc 3061 gaaattgcca gaatcagagt gaaggacatc tcccgcaccg atggtgggag aatgctgatc 3121 cacattggca ggaccaagac cctggtgtcc acagctggtg tggagaaggc cctgtccctg 3181 ggggttacca agctggtgga gagatggatc tctgtgtctg gtgtggctga tgaccccaac 3241 aactacctgt tctgccgggt cagaaagaat ggtgtggctg ccccttctgc cacctcccaa 3301 ctgtccaccc gggccctgga agggatcttt gaggccaccc accgcctgat ctatggtgcc 3361 aaggatgact ctgggcagag atacctggcc tggtctggcc actctgccag agtgggtgct 3421 gccagggaca tggccagggc tggtgtgtcc atccctgaaa tcatgcaggc tggtggctgg 3481 accaatgtga acattgtgat gaactacatc agaaacctgg actctgagac tggggccatg 3541 gtgaggctgc tagaggatgg ggacctcgag ccatctgctg gagacatgag agctgccaac 3601 ctttggccaa gcccgctcat gatcaaacgc tctaagaaga acagcctggc cttgtccctg 3661 acggccgacc agatggtcag tgccttgttg gatgctgagc cccccatact ctattccgag 3721 tatgatccta ccagaccctt cagtgaagct tcgatgatgg gcttactgac caacctggca 3781 gacagggagc tggttcacat gatcaactgg gcgaagaggg tgccaggctt tgtggatttg 3841 accctccatg atcaggtcca ccttctagaa tgtgcctggc tagagatcct gatgattggt 3901 ctcgtctggc gctccatgga gcacccagtg aagctactgt ttgctcctaa cttgctcttg 3961 gacaggaacc agggaaaatg tgtagagggc atggtggaga tcttcgacat gctgctggct 4021 acatcatctc ggttccgcat gatgaatctg cagggagagg agtttgtgtg cctcaaatct 4081 attattttgc ttaattctgg agtgtacaca tttctgtcca gcaccctgaa gtctctggaa 4141 gagaaggacc atatccaccg agtcctggac aagatcacag acactttgat ccacctgatg 4201 gccaaggcag gcctgaccct gcagcagcag caccagcggc tggcccagct cctcctcatc 4261 ctctcccaca tcaggcacat gagtaacaaa ggcatggagc atctgtacag catgaagtgc 4321 aagaacgtgg tgcccctcta tgacctgctg ctggaggcgg cggacgccca ccgcctacat 4381 gcgcccacta gccgtggagg ggcatccgtg gaggagacgg accaaagcca cttggccact 4441 gcgggctcta cttcatcgca ttccttgcaa aagtattaca tcacggggga ggcagagggt 4501 ttccctgcca cagcttgagg cgcgccggat ctttttccct ctgccaaaaa ttatggggac 4561 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca 4621 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat 4681 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct 4741 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc 4801 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt 4861 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt 4921 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc 4981 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 5041 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 5101 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 5161 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta 5221 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 5281 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag 5341 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag 5401 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 5461 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc 5521 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 5581 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 5641 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 5701 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 5761 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 5821 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 5881 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 5941 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 6001 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 6061 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 6121 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 6181 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt 6241 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 6301 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 6361 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 6421 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 6481 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 6541 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 6601 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 6661 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 6721 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 6781 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 6841 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 6901 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 6961 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 7021 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 7081 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 7141 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 7201 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 7261 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 7321 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 7381 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 7441 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 7501 gc //