LOCUS Exported 5423 bp ds-DNA circular SYN 15-JUN-2022 DEFINITION Localization: Protein Expression Vector , Excitation: 399, Emission: 456. ACCESSION . VERSION . KEYWORDS mTagBFP2-pBAD SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5423) AUTHORS Subach OM, Cranfill PJ, Davidson MW, Verkhusha VV TITLE An enhanced monomeric blue fluorescent protein with the high chemical stability of the chromophore. JOURNAL PLoS One. 2011;6(12):e28674. Epub 2011 Dec 8. PUBMED 22174863 REFERENCE 2 (bases 1 to 5423) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 15, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5423 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 80..97 /label=L4440 /note="L4440 vector, forward primer" misc_feature 112..252 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(178..197) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 338..360 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 498..517 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" CDS complement(697..1575) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 1602..1886 /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" primer_bind 1809..1828 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" RBS 1913..1935 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 1955..1972 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 1976..2008 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 2021..2041 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 2048..2758 /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" primer_bind complement(2807..2824) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 2977..3063 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 3155..3182 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 3201..3292 /gene="bla" /label=AmpR promoter CDS 3293..4153 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3511..3530) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4195..4650 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4282..4301) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4492..4513 /label=F1ori-F /note="F1 origin, forward primer" rep_origin 4761..5349 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5250..5269 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg 61 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg 121 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atctggtgca ctctcagtac 181 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg 241 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg 301 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg 361 ttttcaccgt catcaccgaa acgcgcgagg cagaaggaga tggcgcccaa cagtcccccg 421 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga 481 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg 541 ccggtgatgc cggccacgat gcgtccggcg tagaggatct gctcatgttt gacagcttat 601 catcgatgca taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa ccctatgcta 661 ctccgtcaag ccgtcaattg tctgattcgt taccaattat gacaacttga cggctacatc 721 attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt 781 aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat 841 aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca 901 gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa 961 gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat 1021 gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc 1081 ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg 1141 cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg 1201 cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc 1261 atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg 1321 atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca 1381 aacaaattct cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat 1441 ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat 1501 cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt 1561 ttgcgcttca gccatacttt tcatactccc gccattcaga gaagaaacca attgtccata 1621 ttgcatcaga cattgccgtc actgcgtctt ttactggctc ttctcgctaa ccaaaccggt 1681 aaccccgctt attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg 1741 taacaaaagt gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac 1801 actttgctat gccatagcat ttttatccat aagattagcg gatactacct gacgcttttt 1861 atcgcaactc tctactgttt ctccataccc gtttttttgg gctagaaata attttgttta 1921 actttaagaa ggagatatac atatgcgggg ttctcatcat catcatcatc atggtatggc 1981 tagcatgact ggtggacagc aaatgggtcg ggatctgtac gagaacctgt acttccaggg 2041 ctcgagcatg gtgtctaagg gcgaagagct gattaaggag aacatgcaca tgaagctgta 2101 catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc 2161 ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc 2221 cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca 2281 gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac 2341 cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg 2401 cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca 2461 gaagaaaaca ctcggctggg aggccttcac cgagacgctg taccccgctg acggcggcct 2521 ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacgc 2581 caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta 2641 tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca 2701 cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaatta 2761 agaattcgaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta 2821 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg 2881 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg 2941 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg 3001 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca 3061 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga 3121 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt 3181 tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct 3241 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat 3301 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc 3361 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg 3421 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg 3481 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga 3541 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta 3601 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc 3661 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc 3721 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg 3781 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc 3841 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca 3901 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct 3961 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat 4021 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg 4081 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat 4141 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc 4201 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 4261 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 4321 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 4381 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc 4441 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 4501 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga 4561 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 4621 attttaacaa aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat 4681 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 4741 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 4801 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 4861 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag 4921 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 4981 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 5041 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 5101 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 5161 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 5221 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 5281 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 5341 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 5401 gctcacatgt tctttcctgc gtt //