LOCUS Exported 5175 bp ds-DNA circular SYN 17-NOV-2021 DEFINITION Bacterial expression of green fluorescent potassium indicator KRaION1. ACCESSION . VERSION . KEYWORDS pBAD-HisD-KRaION1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5175) AUTHORS Torres Caban CC, Yang M, Lai CC, Yang L, Subach F, Smith BO, Piatkevich KD, Boyden ES TITLE Tuning the sensitivity of genetically encoded fluorescent potassium indicators through structure-guided and genome mining strategies. JOURNAL bioRxiv 2021.10.07.463355 REFERENCE 2 (bases 1 to 5175) AUTHORS . TITLE Direct Submission JOURNAL Exported Nov 17, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5175 /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" primer_bind complement(1576..1593) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 1746..1832 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1924..1951 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 1970..2061 /gene="bla" /label=AmpR promoter CDS 2062..2922 /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(2280..2299) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3093..3681 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3582..3601 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3835..3852 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3867..4007 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3933..3952) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4093..4115 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(4271..5149) /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 atctatggtc 361 tcaaagggag aggaggaaaa catggctagt ctgcctgcta cccacgaact gcatattttt 421 ggctccataa atggcatcga tttcgacatg gtaggccaag gcaccggtaa tccaaacgac 481 ggctatgagg agctgaacct gaagtccacc atgggcgatc tgcagttcag tccctggatc 541 ctggtgccgc acattgggta cgggtttcac cagtatcttc cctatccaga cgggatgtca 601 ccatttcagg ctgcaatggt cgacgggtcc ggataccaag tgcaccgcac tatgcagttt 661 gaggacggtg cctcactgac cgtgaattac agatacacct acgaaggatc acatatcaaa 721 ggggaagctc aggtggaggg cactgggttc ccagctgacg gccctgtgat gacaaattcc 781 ctgacagccg aggcccacgg cctgttcaac ttcgtgaagg acgccggcga gaagctgtgg 841 gatgccgtga caggccagca cgacaaggac gaccaggcca agaaggtgca ggaacacctg 901 aacaagaccg gcatccccga cgccgacaaa gtgaacatcc agatcgccga cggcaaggcc 961 acagtgacag gcgacggact gagccaggaa gccaaagaaa agatcctggt ggccgtgggc 1021 aacatcagcg gaatcgccag cgtggacgat caagtgaaaa ccgccacccc tgccaccgcc 1081 agccagttct acacagtgaa gtccggcgac accctgagcg ccatcagcaa acaggtgtac 1141 ggcaacgcca acctgtacaa caagattttc gaggccaaca agcccatgct gaagtccccc 1201 gacaaaatct accccggcca ggtgctgaga atccccgagg aaatgtactt cgctgactgg 1261 tgcgtatcca agaagacttg ccccaatgat aagactatcg tctccacatt taagtgggct 1321 tttatcaccg acaacggcaa acgctaccgg tctaccgctc gcaccacata cacattcgcc 1381 aagccaatgg ccgcaaatta cctcaaaaat cagccgatgt atgtctttcg gaaaacagag 1441 ctgaaacatt ctaagaccga actgaatttt aaagagtggc aaaaggcctt tacggacgtg 1501 atgggcatgg acgagctgta taagtaataa gaattcgaag cttggctgtt ttggtggatg 1561 agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca 1621 gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt 1681 gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca 1741 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt 1801 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga 1861 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta 1921 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt 1981 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa 2041 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt 2101 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat 2161 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag 2221 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg 2281 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata 2341 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat 2401 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc 2461 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg 2521 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac 2581 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact 2641 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa 2701 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct 2761 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc 2821 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga 2881 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac 2941 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag 3001 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg 3061 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc 3121 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag 3181 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc 3241 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac 3301 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc 3361 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt 3421 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt 3481 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc 3541 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt 3601 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca 3661 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt 3721 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt 3781 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag 3841 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc 3901 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 3961 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc 4021 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag 4081 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg 4141 cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca tgcataatgt gcctgtcaaa 4201 tggacgaagc agggattctg caaaccctat gctactccgt caagccgtca attgtctgat 4261 tcgttaccaa ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac 4321 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat 4381 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca 4441 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct 4501 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga 4561 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat 4621 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct 4681 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga 4741 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg 4801 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt 4861 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc 4921 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca 4981 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt 5041 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg 5101 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac 5161 tcccgccatt cagag //