LOCUS       sequence-207270-        7417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression plasmid for potassium ion ratiometric 
            indicator KIRIN1 with nuclear exclusion sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7417)
  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 7417)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2019-01-14"; volume: "2:18. doi"; pages: " 
            10.1038/s42003-018-0269-2. eCollection 2019"
COMMENT     Sequence Label: pcDNA-NES-KIRIN1
FEATURES             Location/Qualifiers
     source          1..7417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        157..536
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        537..740
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     691..711
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     785..804
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        785..803
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(904..925)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1165..1184)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2824..2853
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2869..2886
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2930..2947)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2936..3160
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3206..3634
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3293..3312)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3503..3524
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3643..3663)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3648..3977
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3828..3963
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3890..3909
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4044..4838
                     /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(4098..4117)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4708..4727
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5012..5133
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5049..5068)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5103..5122
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5182..5198)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5182..5198)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5195..5217)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5206..5222
                     /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(join(5230..5236,5237..5254,5255..5260))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5275..5296
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5413..5430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5584..6172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5664..5683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6343..7134,7135..7203))
                     /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     6966..6985
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7204..7308)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7383..7402)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
       61 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
      121 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
      181 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      241 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      301 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      361 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      421 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      481 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      541 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      601 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      661 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      721 aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg
      781 aaattaatac gactcactat agggagaccc aagctggcta gcatgttagc cttgaaatta
      841 gcaggtcttg atatcgggag cgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      901 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      961 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1021 cccgtgccct ggcccaccct cgtgaccacc ctgagctggg gcgtgcagtg cttcgcccgc
     1081 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1141 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1201 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1261 ggcaacatcc tggggcacaa gctggagtac aacgccatcc acggcaacgt ctatatcacc
     1321 gccgacaagc agaagaacgg catcaaggcc aacttcggcc tcaactgcaa catcgaggac
     1381 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1441 ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag
     1501 aagcgcgatc acatggtcct gctggagttc gtgaccgcgg ccgcaggcct gttcaacttc
     1561 gtgaaggacg ccggcgagaa gctgtgggat gccgtgacag gccagcacga caaggacgac
     1621 caggccaaga aggtgcagga acacctgaac aagaccggca tccccgacgc cgacaaagtg
     1681 aacatccaga tcgccgacgg caaggccaca gtgacaggcg acggactgag ccaggaagcc
     1741 aaagaaaaga tcctggtggc cgtgggcaac atcagcggaa tcgccagcgt ggacgatcaa
     1801 gtgaaaaccg ccacccctgc caccgccagc cagttctaca cagtgaagtc cggcgacacc
     1861 ctgagcgcca tcagcaaaca ggtgtacggc aacgccaacc tgtacaacaa gattttcgag
     1921 gccaacaagc ccatgctgaa gtcccccgac aaaatctacc ccggccaggt gctgagaatc
     1981 cccgaggaac tggagctcat gggtggggtc cagcttgcag atcactacca gcagaatact
     2041 ccgatcgggg atgggccggt tctcctgcct gacaaccact acctgtcata ccagtccaag
     2101 ctgtccaaag atcctaatga aaagcgggat cacatggtcc tgctggaatt tgtcaccgcc
     2161 gccggaatca ctctcggaat ggacgagctg tacaaaggtg ggaccggggg atctatggtg
     2221 agcaagggcg aagaattgtt cactggtgta gtccctattt tggtggaact ggatggggat
     2281 gtcaacggtc ataaattcag cgtgcgaggg gagggtgagg gggacgcaac taatggcaaa
     2341 ttgacactca agctcatctg cacaacgggg aaactgcccg tgccgtggcc cacactcgtt
     2401 actactctag gatatggcct tatgtgtttc gcccgctacc cagaccacat gaaacaacac
     2461 gatttcttta aatcagccat gccagagggg tacgtccagg agagaactat cttcttcaag
     2521 gacgatggta attacaagac tagggccgag gtgaaattcg aaggggatac acttgtcaac
     2581 cgaatcgagc tgaagggaat cgactttaag gaggacggaa acatactcgg gcataagttg
     2641 gaatacaact acaacagtca caatgtttac atcacagccg ataagcaaaa gaacggcatt
     2701 aaggctaatt ttaagatccg tcacaacatt gaggattaac tcgagcggcc gccactgtgc
     2761 tggatatctg cagaattcca ccacactgga ctagtggatc cgagctcggt accaagcttt
     2821 ctagaacaaa aactcatctc agaagaggat ctgaatagcg ccgtcgacca tcatcatcat
     2881 catcattgag tttaaacggt ctccagctta agtttaaacc gctgatcagc ctcgactgtg
     2941 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3001 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3061 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3121 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3181 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3241 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3301 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3361 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3421 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3481 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3541 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3601 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3661 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3721 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3781 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3841 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3901 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3961 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4021 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4081 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4141 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4201 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4261 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4321 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4381 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4441 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4501 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4561 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4621 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4681 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4741 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4801 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4861 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4921 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4981 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5041 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5101 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5161 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5221 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5281 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5341 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5401 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5461 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5521 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5581 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5641 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5701 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5761 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5821 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5881 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5941 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6001 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6061 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6121 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6181 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6241 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6301 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6361 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6421 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6481 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6541 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6601 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6661 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6721 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6781 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6841 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6901 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6961 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7021 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7081 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7141 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7201 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7261 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7321 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     7381 agtacaatct gctctgatgc cgcatagtta agccagt
//
