LOCUS Exported 7288 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian cell expression plasmid for potassium ion raitometric indicator KIRIN1-GR. ACCESSION . VERSION . KEYWORDS pcDNA-KIRIN1-GR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7288) 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 7288) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7288 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 177..556 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 557..760 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 711..731 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 805..824 /label=T7 /note="T7 promoter, forward primer" promoter 805..823 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(915..936) /label=EGFP-N /note="EGFP, reverse primer" primer_bind 1523..1544 /label=EGFP-C /note="EGFP, forward primer" CDS 2715..2744 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2760..2777 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2821..2838) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2827..3051 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3097..3525 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3184..3203) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3394..3415 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3534..3554) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3539..3868 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3719..3854 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3781..3800 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3935..4729 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3989..4008) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4599..4618 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4903..5024 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4940..4959) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4994..5013 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5073..5089) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5073..5089) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5086..5108) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5097..5113 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5121..5151) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5166..5187 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5304..5321) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5475..6063) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5555..5574) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6234..7094) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6857..6876 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7095..7199) /gene="bla" /label=AmpR promoter primer_bind complement(7274..5) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 61 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 121 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 181 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 241 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 301 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 361 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 421 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 481 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 541 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 601 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 661 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 721 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc 781 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta 841 gcgtttaaac gggccctcta gactcgagca tggtgagcaa gggcgaggag ctgttcaccg 901 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtcc 961 gcggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca 1021 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcgtggcct 1081 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg 1141 aaggctacgt ccaggagcgc accatctctt tcaaggacga cggtacctac aagacccgcg 1201 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact 1261 tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac agccacaacg 1321 tctatatcac ggccgacaag cagaagaacg gcatcaaggc taacttcaag atccgccaca 1381 acgttgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 1441 acggccccgt gctgctgccc gacaaccact acctgagcca tcagtccgcc ctgagcaaag 1501 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgtcgacg 1561 gcctgttcaa cttcgtgaag gacgccggcg agaagctgtg ggatgccgtg acaggccagc 1621 acgacaagga cgaccaggcc aagaaggtgc aggaacacct gaacaagacc ggcatccccg 1681 acgccgacaa agtgaacatc cagatcgccg acggcaaggc cacagtgaca ggcgacggac 1741 tgagccagga agccaaagaa aagatcctgg tggccgtggg caacatcagc ggaatcgcca 1801 gcgtggacga tcaagtgaaa accgccaccc ctgccaccgc cagccagttc tacacagtga 1861 agtccggcga caccctgagc gccatcagca aacaggtgta cggcaacgcc aacctgtaca 1921 acaagatttt cgaggccaac aagcccatgc tgaagtcccc cgacaaaatc taccccggcc 1981 aggtgctgag aatccccgag gaactggaat tcatcaagga aaatatgcgt atgaaggtgg 2041 tcatggaagg ttcggtcaac ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc 2101 cgtacatggg aactcaaacc atgaggatca aagtcatcga gggaggaccc ctgccatttg 2161 cctttgacat tcttgccacg tcgttcatgt atggcagccg tacttttatc aagtacccga 2221 aaggcattcc tgatttcttt aaacagtcct ttcctgaggg ttttacttgg gaaagagtta 2281 cgagatacga agatggtgga gtcgtcaccg tcatgcagga caccagcctt gaggatggct 2341 gtctcgttta ccacgtccaa gtcagagggg taaactttcc ctccaatggt cccgtgatgc 2401 agaagaagac caagggttgg gagcctaata cagagatgat gtatccagca gatggtggtc 2461 tgaggggata cactcatatg gcactgaaag ttgatggtgg tggccatctg tcttgctctt 2521 tcgtaacaac ttacaggtca aaaaagaccg tcgggaacat caagatgccc ggtatccatg 2581 ccgttgatca ccgcctggaa aggttagagg aaagtgacaa tgaaatgttc gtagtacaac 2641 gcgaacacgc agttgccaag ttcgccgggc ttggtggtgg gatggacgag ctgtacaagt 2701 aaaagcttgg gcccgaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc 2761 atcatcatca tcatcattga gtttaaacgg tctccagctt aagtttaaac cgctgatcag 2821 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 2881 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 2941 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 3001 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 3061 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa 3121 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 3181 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 3241 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 3301 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 3361 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 3421 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 3481 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt 3541 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 3601 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 3661 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 3721 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 3781 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg 3841 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg 3901 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 3961 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 4021 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 4081 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg 4141 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 4201 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 4261 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 4321 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 4381 agccggcctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 4441 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg 4501 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 4561 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 4621 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 4681 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 4741 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 4801 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 4861 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct 4921 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 4981 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg 5041 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 5101 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 5161 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 5221 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 5281 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 5341 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 5401 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 5461 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 5521 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 5581 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 5641 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 5701 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 5761 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 5821 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 5881 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 5941 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 6001 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 6061 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 6121 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 6181 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 6241 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 6301 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 6361 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 6421 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 6481 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 6541 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 6601 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 6661 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 6721 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 6781 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 6841 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 6901 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 6961 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 7021 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 7081 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 7141 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 7201 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat 7261 ggtgcactct cagtacaatc tgctctga //