LOCUS Exported 4710 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBAD-PSmOrange2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4710) AUTHORS Subach OM, Entenberg D, Condeelis JS, Verkhusha VV TITLE A FRET-facilitated photoswitching using an orange fluorescent protein with the fast photoconversion kinetics. JOURNAL J Am Chem Soc. 2012 Aug 17. PUBMED 22900938 REFERENCE 2 (bases 1 to 4710) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4710 /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 328..345 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 355..1062 /codon_start=1 /product="enhanced orange to far-red photoswitchable monomeric fluorescent protein (Subach et al., 2012)" /label=PSmOrange2 /note="mammalian codon-optimized" /translation="MVSKGEENNMAIIKEFMRFKVHMEGTVNGHEFEIEGEGEGHPYEG FQTAKLKVTKGGPLPFAWDILSPLITYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY EDGGVVTVTQDSSLQDGEFIYKVKMRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIRMRLKLKDGGHYTSEVKTTYKAKKSVLLPGAYIVGIKLDITSHNEDYTIVEQYERS EARHSTGGMDELYK" primer_bind 768..787 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(1111..1128) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 1281..1367 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1459..1486 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 1505..1596 /gene="bla" /label=AmpR promoter CDS 1597..2457 /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(1815..1834) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2628..3216 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3117..3136 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3370..3387 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3402..3542 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3468..3487) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3628..3650 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3806..4684) /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 ggggagccac catcaccatc accatggcag atctatggtg 361 agcaagggcg aggagaataa tatggccatc attaaggagt tcatgcgctt caaggtgcac 421 atggagggca ctgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccacccc 481 tacgagggct ttcagaccgc taagctgaag gtgaccaagg gtggccccct gcccttcgcc 541 tgggacatcc tgtcccctct catcacctac ggctccaagg cgtacgtgaa gcaccccgcc 601 gacatccccg actacttcaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg 661 aactacgagg acggcggcgt ggtgaccgtg acccaggact cctctctgca ggacggcgag 721 ttcatctaca aggtgaagat gcgcggcacc aacttcccct ccgacggccc cgtaatgcag 781 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg 841 aagggcgaga tcaggatgag gctgaagctg aaggatggcg gccactacac ctccgaggtc 901 aagactacct acaaggccaa gaagtccgtg ctgctgcccg gcgcctacat cgtcggcatc 961 aagttggaca ttacctcaca caacgaggac tacaccatcg tggaacagta tgaacgcagc 1021 gaggcccgcc actccaccgg cggcatggac gagctgtaca agtaagaatt cgaagcttgg 1081 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag 1141 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat 1201 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag 1261 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc 1321 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg 1381 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg 1441 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa 1501 ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc 1561 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc 1621 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg 1681 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat 1741 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc 1801 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa 1861 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa 1921 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt 1981 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct 2041 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat 2101 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg 2161 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg 2221 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt 2281 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg 2341 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg 2401 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg 2461 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 2521 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt 2581 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 2641 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 2701 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 2761 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta 2821 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 2881 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 2941 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 3001 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 3061 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 3121 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 3181 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 3241 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat 3301 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg 3361 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc 3421 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct 3481 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg 3541 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat 3601 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt 3661 catcaccgaa acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag cggcatgcat 3721 aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc 3781 cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt 3841 cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg 3901 agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg 3961 tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc 4021 taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct 4081 gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag 4141 cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc 4201 gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt 4261 gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg 4321 ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg 4381 cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt 4441 agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc 4501 gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca 4561 ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac 4621 ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag 4681 ccatactttt catactcccg ccattcagag //