LOCUS Exported 5897 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial Expression plasmid for potassium ion raitometric indicator KIRIN1-GR. ACCESSION . VERSION . KEYWORDS pBAD-KIRIN1-GR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5897) AUTHORS Shen Y, Wu SY, Rancic V, Aggarwal A, Qian Y, Miyashita SI, Ballanyi K, Campbell RE, Dong M TITLE Genetically encoded fluorescent indicators for imaging intracellular potassium ion concentration. JOURNAL Commun Biol. 2019 Jan 14;2:18. doi: 10.1038/s42003-018-0269-2. eCollection 2019. PUBMED 30652129 REFERENCE 2 (bases 1 to 5897) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5897 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 17..301 /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 224..243 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" CDS 347..364 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 368..400 /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 404..427 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" primer_bind complement(488..509) /label=EGFP-N /note="EGFP, reverse primer" primer_bind 1096..1117 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(2314..2331) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 2484..2570 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2662..2689 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 2708..2799 /gene="bla" /label=AmpR promoter CDS 2800..3660 /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(3018..3037) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3831..4419 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4320..4339 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4573..4590 /label=L4440 /note="L4440 vector, forward primer" misc_feature 4605..4745 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4671..4690) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4831..4853 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(5009..5887) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" ORIGIN 1 ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc 61 gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg 121 taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc 181 agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta 241 tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca 301 tacccgtttt ttgggctaac aggaggaatt aaccatgggg ggttctcatc atcatcatca 361 tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga 421 cgataaggat ccgagctcga gcatggtgag caagggcgag gagctgttca ccggggtggt 481 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tccgcggcga 541 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa 601 gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcgtgg cctgcttcag 661 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 721 cgtccaggag cgcaccatct ctttcaagga cgacggtacc tacaagaccc gcgccgaggt 781 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 841 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca acgtctatat 901 cacggccgac aagcagaaga acggcatcaa ggctaacttc aagatccgcc acaacgttga 961 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 1021 cgtgctgctg cccgacaacc actacctgag ccatcagtcc gccctgagca aagaccccaa 1081 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccgtcg acggcctgtt 1141 caacttcgtg aaggacgccg gcgagaagct gtgggatgcc gtgacaggcc agcacgacaa 1201 ggacgaccag gccaagaagg tgcaggaaca cctgaacaag accggcatcc ccgacgccga 1261 caaagtgaac atccagatcg ccgacggcaa ggccacagtg acaggcgacg gactgagcca 1321 ggaagccaaa gaaaagatcc tggtggccgt gggcaacatc agcggaatcg ccagcgtgga 1381 cgatcaagtg aaaaccgcca cccctgccac cgccagccag ttctacacag tgaagtccgg 1441 cgacaccctg agcgccatca gcaaacaggt gtacggcaac gccaacctgt acaacaagat 1501 tttcgaggcc aacaagccca tgctgaagtc ccccgacaaa atctaccccg gccaggtgct 1561 gagaatcccc gaggaactgg aattcatcaa ggaaaatatg cgtatgaagg tggtcatgga 1621 aggttcggtc aacggccacc aattcaaatg cacaggtgaa ggagaaggca atccgtacat 1681 gggaactcaa accatgagga tcaaagtcat cgagggagga cccctgccat ttgcctttga 1741 cattcttgcc acgtcgttca tgtatggcag ccgtactttt atcaagtacc cgaaaggcat 1801 tcctgatttc tttaaacagt cctttcctga gggttttact tgggaaagag ttacgagata 1861 cgaagatggt ggagtcgtca ccgtcatgca ggacaccagc cttgaggatg gctgtctcgt 1921 ttaccacgtc caagtcagag gggtaaactt tccctccaat ggtcccgtga tgcagaagaa 1981 gaccaagggt tgggagccta atacagagat gatgtatcca gcagatggtg gtctgagggg 2041 atacactcat atggcactga aagttgatgg tggtggccat ctgtcttgct ctttcgtaac 2101 aacttacagg tcaaaaaaga ccgtcgggaa catcaagatg cccggtatcc atgccgttga 2161 tcaccgcctg gaaaggttag aggaaagtga caatgaaatg ttcgtagtac aacgcgaaca 2221 cgcagttgcc aagttcgccg ggcttggtgg tgggatggac gagctgtaca agtaaaagct 2281 tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag 2341 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc 2401 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc 2461 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct 2521 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag 2581 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa 2641 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac 2701 aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 2761 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 2821 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 2881 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 2941 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 3001 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag 3061 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 3121 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 3181 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 3241 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 3301 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 3361 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 3421 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 3481 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 3541 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 3601 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 3661 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 3721 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 3781 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 3841 ttttttctgc gcgtaatctg ctgcttgcaa acaaaacaac caccgctacc agcggtggtt 3901 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 3961 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct 4021 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 4081 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 4141 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 4201 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 4261 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 4321 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 4381 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 4441 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 4501 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga 4561 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt 4621 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc 4681 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg 4741 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg 4801 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac 4861 cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg 4921 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca 4981 agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt 5041 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc 5101 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc 5161 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga 5221 cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat 5281 gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac 5341 aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc 5401 gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc 5461 cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat 5521 ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt 5581 aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac 5641 cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt 5701 ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt 5761 tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta 5821 aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt 5881 cagccatact tttcata //