LOCUS Exported 4785 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBAD/HisB-PAmCherry1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4785) AUTHORS Subach FV, Patterson GH, Manley S, Gillette JM, Lippincott-Schwartz J, Verkhusha VV TITLE Photoactivatable mCherry for high-resolution two-color fluorescence microscopy. JOURNAL Nat Methods. 2009 Feb;6(2):153-9. Epub 2009 Jan 25. PUBMED 19169259 REFERENCE 2 (bases 1 to 4785) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4785 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..285 /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 208..227 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" CDS 331..348 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 352..384 /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 388..411 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 430..1140 /codon_start=1 /product="photoactivatable monomeric derivative of DsRed fluorescent protein" /label=PAmCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(575..593) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 843..862 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(1186..1203) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 1356..1442 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1534..1561 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 1580..1671 /gene="bla" /label=AmpR promoter CDS 1672..2532 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1890..1909) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2703..3291 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3192..3211 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3445..3462 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3477..3617 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3543..3562) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3703..3725 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3881..4759) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" ORIGIN 1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct 61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca 121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg 181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg 241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc 301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc 361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc 421 tcgagatcta tggtgagcaa gggcgaggag gataacatgg ccatcattaa ggagttcatg 481 cgcttcaagg tgcacatgga ggggtccgtg aacggccacg tgttcgagat cgagggcgag 541 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc 601 cccctgccct tcacctggga catcctgtcc cctcaattca tgtacggctc caatgcctac 661 gtgaagcacc ccgccgacat ccccgactac tttaagctgt ccttccccga gggcttcaag 721 tgggagcgcg tgatgaaatt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc 781 ctgcaggacg gtgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac 841 ggccccgtaa tgcagaagaa gaccatgggc tgggaggccc tctccgagcg gatgtacccc 901 gaggacggcg ccctgaaggg cgaggtcaag ccgagagtga agctgaagga cggcggccac 961 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc 1021 tacaacgtca accgcaagtt ggacatcacc tcacacaacg aggactacac catcgtggaa 1081 cagtacgaac gtgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa 1141 gaattcgaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa 1201 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt 1261 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg 1321 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga 1381 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa 1441 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac 1501 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt 1561 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc 1621 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 1681 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 1741 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 1801 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 1861 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac 1921 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 1981 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 2041 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 2101 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 2161 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 2221 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 2281 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 2341 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 2401 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 2461 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 2521 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 2581 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 2641 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 2701 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 2761 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 2821 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac 2881 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 2941 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 3001 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 3061 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 3121 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 3181 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 3241 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 3301 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 3361 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct 3421 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg 3481 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc 3541 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg 3601 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt 3661 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc 3721 agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg 3781 cgaagcggca tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat 3841 gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta 3901 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt 3961 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg 4021 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc 4081 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg 4141 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc 4201 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa 4261 tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat 4321 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct 4381 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc 4441 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct 4501 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc 4561 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga 4621 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct 4681 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat 4741 cattttgcgc ttcagccata cttttcatac tcccgccatt cagag //