LOCUS Exported 6586 bp ds-DNA circular SYN 24-MAR-2023 DEFINITION Monomeric TagBFP2 and iCre recombinase (version 2). ACCESSION . VERSION . KEYWORDS pCAGGS-mTagBFP2-T2A-iCre (v2) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6586) AUTHORS Nam HS, Capecchi MR TITLE 2A plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 6586) AUTHORS . TITLE Direct Submission JOURNAL Exported Mar 24, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6586 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 11..390 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 392..669 /label=chicken beta-actin promoter intron 670..1687 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1611..1633 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1695..1714 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1745..2455 /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 2477..2530 /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 2552..2572 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(3616..3635) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3681..3736 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3735..3754) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4097..4113) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4097..4113) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4110..4132) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4121..4137 /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(4145..4175) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4189..4210 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4269..4464 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4315..4450 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4377..4396 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4470..4604 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4507..4526) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4561..4580 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4672..4689) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4842..5430) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4922..4941) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5601..6461) /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 6224..6243 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6462..6566) /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 gggccccccc caggcggggc ggggcggggc 541 gaggggcggg gcggggcgag gcggaaaggt gcggcggcag ccaatcagag cggcgcgctc 601 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg 661 cggcgggcgg gagtcgttgc gcgctgcctt ccccccgtgc 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 ggccggggag ggctcggggg aaggggcgcg gcggcccccg 1321 gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg 1381 cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg 1441 ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat 1501 gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg 1561 gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt 1621 ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt 1681 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt 1741 cgccatggtg tctaagggcg aagagctgat taaggagaac atgcacatga agctgtacat 1801 ggagggcacc gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta 1861 cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt 1921 cgacatcctg gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg 1981 catccccgac ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac 2041 atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct 2101 catctacaac gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa 2161 gaaaacactc ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga 2221 aggcagaaac gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacgccaa 2281 gaccacatat agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt 2341 ggactacaga ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga 2401 ggtggcagtg gccagatact gcgacctccc tagcaaactg gggcacaagc ttaatggcgg 2461 ctccggcggc cgctccgagg gccgcggctc cctgctgacc tgcggcgacg tggaggagaa 2521 ccccggcccc gcccccggct ccaccatggt gcccaagaag aagaggaaag tctccaacct 2581 gctgactgtg caccaaaacc tgcctgccct ccctgtggat gccacctctg atgaagtcag 2641 gaagaacctg atggacatgt tcagggacag gcaggccttc tctgaacaca cctggaagat 2701 gctcctgtct gtgtgcagat cctgggctgc ctggtgcaag ctgaacaaca ggaaatggtt 2761 ccctgctgaa cctgaggatg tgagggacta cctcctgtac ctgcaagcca gaggcctggc 2821 tgtgaagacc atccaacagc acctgggcca gctcaacatg ctgcacagga gatctggcct 2881 gcctcgccct tctgactcca atgctgtgtc cctggtgatg aggagaatca gaaaggagaa 2941 tgtggatgct ggggagagag ccaagcaggc cctggccttt gaacgcactg actttgacca 3001 agtcagatcc ctgatggaga actctgacag atgccaggac atcaggaacc tggccttcct 3061 gggcattgcc tacaacaccc tgctgcgcat tgccgaaatt gccagaatca gagtgaagga 3121 catctcccgc accgatggtg ggagaatgct gatccacatt ggcaggacca agaccctggt 3181 gtccacagct ggtgtggaga aggccctgtc cctgggggtt accaagctgg tggagagatg 3241 gatctctgtg tctggtgtgg ctgatgaccc caacaactac ctgttctgcc gggtcagaaa 3301 gaatggtgtg gctgcccctt ctgccacctc ccaactgtcc acccgggccc tggaagggat 3361 ctttgaggcc acccaccgcc tgatctatgg tgccaaggat gactctgggc agagatacct 3421 ggcctggtct ggccactctg ccagagtggg tgctgccagg gacatggcca gggctggtgt 3481 gtccatccct gaaatcatgc aggctggtgg ctggaccaat gtgaacattg tgatgaacta 3541 catcagaaac ctggactctg agactggggc catggtgagg ctgctcgagg atggggactg 3601 ataaggcgcg ccggatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct 3661 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa 3721 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa 3781 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg 3841 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca 3901 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc 3961 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg 4021 actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa 4081 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 4141 cacacaacat acgagccgga agcataaagt gtaaagcctg ggtgcctaat gagtgagcta 4201 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca 4261 gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg 4321 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 4381 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt 4441 tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca 4501 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 4561 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat 4621 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 4681 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 4741 aatacggtta tccacagaat cagggataac gcaggaaaga acatgtgagc aaaaggccag 4801 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 4861 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 4921 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 4981 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 5041 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 5101 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 5161 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 5221 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 5281 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 5341 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 5401 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 5461 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 5521 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 5581 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 5641 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 5701 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 5761 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 5821 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 5881 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 5941 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 6001 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 6061 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 6121 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 6181 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 6241 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 6301 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 6361 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 6421 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 6481 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 6541 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgc //