LOCUS Exported 6589 bp ds-DNA circular SYN 12-MAY-2021 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ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6589) AUTHORS Nam HS, Capecchi MR TITLE 2A plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 6589) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6589 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 11..390 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 672..1688 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1612..1634 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1696..1715 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1746..2456 /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 2478..2531 /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 2553..2573 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(3617..3636) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3682..3737 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3736..3755) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4098..4114) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4098..4114) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4111..4133) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4122..4138 /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(4146..4176) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4191..4212 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4271..4466 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4317..4452 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4379..4398 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4472..4606 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4509..4528) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4563..4582 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4674..4691) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4845..5433) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4925..4944) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5604..6464) /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 6227..6246 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6465..6569) /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 gggsgcgcgc nncaggcggg gcggggcggg 541 gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc 601 tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg 661 cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct 721 cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg 781 gcccttctcc tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt 841 ggctgcgtga aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg 901 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg 961 gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga 1021 gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg 1081 tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac 1141 cccccctgca cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc 1201 gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc 1261 cgggcggggc ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc 1321 ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt 1381 gcgagagggc gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc 1441 gccgccgcac cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa 1501 tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc 1561 ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct 1621 tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt 1681 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat 1741 tcgccatggt gtctaagggc gaagagctga ttaaggagaa catgcacatg aagctgtaca 1801 tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct 1861 acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct 1921 tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg 1981 gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca 2041 catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc 2101 tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga 2161 agaaaacact cggctgggag gccttcaccg agacgctgta ccccgctgac ggcggcctgg 2221 aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacgcca 2281 agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg 2341 tggactacag actggaaaga atcaaggagg ccaacaacga gacctacgtc gagcagcacg 2401 aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag cttaatggcg 2461 gctccggcgg ccgctccgag ggccgcggct ccctgctgac ctgcggcgac gtggaggaga 2521 accccggccc cgcccccggc tccaccatgg tgcccaagaa gaagaggaaa gtctccaacc 2581 tgctgactgt gcaccaaaac ctgcctgccc tccctgtgga tgccacctct gatgaagtca 2641 ggaagaacct gatggacatg ttcagggaca ggcaggcctt ctctgaacac acctggaaga 2701 tgctcctgtc tgtgtgcaga tcctgggctg cctggtgcaa gctgaacaac aggaaatggt 2761 tccctgctga acctgaggat gtgagggact acctcctgta cctgcaagcc agaggcctgg 2821 ctgtgaagac catccaacag cacctgggcc agctcaacat gctgcacagg agatctggcc 2881 tgcctcgccc ttctgactcc aatgctgtgt ccctggtgat gaggagaatc agaaaggaga 2941 atgtggatgc tggggagaga gccaagcagg ccctggcctt tgaacgcact gactttgacc 3001 aagtcagatc cctgatggag aactctgaca gatgccagga catcaggaac ctggccttcc 3061 tgggcattgc ctacaacacc ctgctgcgca ttgccgaaat tgccagaatc agagtgaagg 3121 acatctcccg caccgatggt gggagaatgc tgatccacat tggcaggacc aagaccctgg 3181 tgtccacagc tggtgtggag aaggccctgt ccctgggggt taccaagctg gtggagagat 3241 ggatctctgt gtctggtgtg gctgatgacc ccaacaacta cctgttctgc cgggtcagaa 3301 agaatggtgt ggctgcccct tctgccacct cccaactgtc cacccgggcc ctggaaggga 3361 tctttgaggc cacccaccgc ctgatctatg gtgccaagga tgactctggg cagagatacc 3421 tggcctggtc tggccactct gccagagtgg gtgctgccag ggacatggcc agggctggtg 3481 tgtccatccc tgaaatcatg caggctggtg gctggaccaa tgtgaacatt gtgatgaact 3541 acatcagaaa cctggactct gagactgggg ccatggtgag gctgctcgag gatggggact 3601 gataaggcgc gccggatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc 3661 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga 3721 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga 3781 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag 3841 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc 3901 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt 3961 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct 4021 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca 4081 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 4141 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 4201 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 4261 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 4321 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 4381 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct 4441 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta 4501 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 4561 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga 4621 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 4681 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 4741 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 4801 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 4861 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 4921 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 4981 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 5041 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 5101 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 5161 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 5221 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 5281 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 5341 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 5401 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 5461 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 5521 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 5581 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 5641 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 5701 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 5761 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 5821 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 5881 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 5941 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 6001 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 6061 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 6121 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 6181 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 6241 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 6301 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 6361 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 6421 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 6481 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 6541 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgc //