LOCUS Exported 5175 bp ds-DNA circular SYN 04-JAN-2022 DEFINITION Bacterial expression of green fluorescent potassium indicator mNG-C-Kbp. ACCESSION . VERSION . KEYWORDS pBAD-HisD-mNG-C-Kbp 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 Jan 4, 2022 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 40..324 /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 247..266 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" CDS 367..384 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1615..1632) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 1785..1871 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1963..1990 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 2009..2100 /gene="bla" /label=AmpR promoter CDS 2101..2961 /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(2319..2338) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3132..3720 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3621..3640 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3874..3891 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3906..4046 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3972..3991) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4132..4154 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(4310..13) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" ORIGIN 1 gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat tgtccatatt 61 gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc aaaccggtaa 121 ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta 181 acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac 241 tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga cgctttttat 301 cgcaactctc tactgtttct ccatacccgt tttttgggct aacaggagga attaaccatg 361 gggagccacc atcaccatca ccatggcaga tctatggtct caaagggaga ggaggaaaac 421 atggctagtc tgcctgctac ccacgaactg catatttttg gctccataaa tggcatcgat 481 ttcgacatgg taggccaagg caccggtaat ccaaacgacg gctatgagga gctgaacctg 541 aagtccacca tgggcgatct gcagttcagt ccctggatcc tggtgccgca cattgggtac 601 gggtttcacc agtatcttcc ctatccagac gggatgtcac catttcaggc tgcaatggtc 661 gacgggtccg gataccaagt gcaccgcact atgcagtttg aggacggtgc ctcactgacc 721 gtgaattaca gatacaccta cgaaggatca catatcaaag gggaagctca ggtggagggc 781 actgggttcc cagctgacgg ccctgtgatg acaaattccc tgacagccga ggcccacatg 841 ggcctgttca acttcgtgaa ggaggccggc gagaagctgt gggacaccgt gaccggcaac 901 gccaccgccg aggaccagag cgccaagctg aaggaccacc tgaacaagac cggcctgccc 961 ggcatcgaca aggtggacgt gcaggtggtg gacggcaagg ccgtggtgac cggcgacgcc 1021 gtgagccagg agctgaagga gaagatcctg gtggccgtgg gcaacgtggc cggcatcagc 1081 ggcgtggagg acaaggtgag cgtggagcag gccgccgccg agagcaggtt ctacaccgtg 1141 aagaagggcg acaccctgag cgccgtgagc aaggaggtgt acggcaacgc caacctgtac 1201 aacaagatct tcgaggccaa caagcccatg ctgagcagcc ccgacaagat ctaccccggc 1261 caggtgctga ggatccccga gatgtacttc gctgactggt gcgtatccaa gaagacttgc 1321 cccaatgata agactatcgt ctccacattt aagtgggctt ttatcaccga caacggcaaa 1381 cgctaccggt ctaccgctcg caccacatac acattcgcca agccaatggc cgcaaattac 1441 ctcaaaaatc agccgatgta tgtctttcgg aaaacagagc tgaaacattc taagaccgaa 1501 ctgaatttta aagagtggca aaaggccttt acggacgtga tgggcatgga cgagctgtat 1561 aagtaataag aattcgaagc ttggctgttt tggtggatga gagaagattt tcagcctgat 1621 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag 1681 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg 1741 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg 1801 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga 1861 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc 1921 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg 1981 atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg 2041 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 2101 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 2161 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca 2221 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 2281 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 2341 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 2401 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 2461 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc 2521 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 2581 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 2641 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct 2701 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 2761 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct 2821 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 2881 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 2941 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat 3001 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 3061 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 3121 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 3181 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 3241 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta 3301 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 3361 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 3421 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 3481 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 3541 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 3601 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 3661 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 3721 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 3781 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 3841 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 3901 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg 3961 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat 4021 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct 4081 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 4141 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc 4201 gcgcgaaggc gaagcggcat gcataatgtg cctgtcaaat ggacgaagca gggattctgc 4261 aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact 4321 tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc 4381 cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac 4441 cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt 4501 ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac 4561 gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca 4621 ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg 4681 actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca 4741 gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga 4801 aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc 4861 agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt 4921 cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat 4981 cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg 5041 tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac 5101 cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca 5161 gcaggggatc atttt //