LOCUS       sequence-207096-        5897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for potassium ion ratiometric indicator
            KIRIN1-GR.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 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: pBAD-KIRIN1-GR
FEATURES             Location/Qualifiers
     source          1..5897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(404..412,413..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             join(2800..2868,2869..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
//
