LOCUS       sequence-348254-        5175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of green fluorescent potassium indicator 
            KRaION2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5175)
  AUTHORS   Torres Caban CC, Yang M, Lai C, Yang L, Subach FV, Smith BO, 
            Piatkevich KD, Boyden ES
  TITLE     Tuning the Sensitivity of Genetically Encoded Fluorescent Potassium 
            Indicators through Structure-Guided and Genome Mining Strategies.
  JOURNAL   ACS Sens. 2022 May 27;7(5):1336-1346. doi: 
            10.1021/acssensors.1c02201. Epub 2022 Apr 15.
  PUBMED    35427452
REFERENCE   2  (bases 1 to 5175)
  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"; doi: 
            "10.1021/acssensors.1c02201"; journalName: "ACS Sens"; date: 
            "2022-05-27- 27"; volume: "7"; issue: "5"; pages: "1336-1346"
COMMENT     Sequence Label: pBAD-HisD-KRaION2
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             328..345
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1576..1593)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1746..1832
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1924..1951
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1970..2061
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2062..2130,2131..2922)
                     /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(2280..2299)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3093..3681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3582..3601
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3835..3852
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3867..4007
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3933..3952)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4093..4115
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4271..5149)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggagccac catcaccatc accatggcag atctatggtc
      361 tcaaagggag aggaggaaaa catggctagt ctgcctgcta cccacgaact gcatattttt
      421 ggctccataa atggcatcga tttcgacatg gtaggccaag gcaccggtaa tccaaacgac
      481 ggctatgagg agctgaacct gaagtccacc atgggcgatc tgcagttcag tccctggatc
      541 ctggtgccgc acattgggta cgggtttcac cagtatcttc cctatccaga cgggatgtca
      601 ccatttcagg ctgcaatggt cgacgggtcc ggataccaag tgcaccgcac tatgcagttt
      661 gaggacggtg cctcactgac cgtgaattac agatacacct acgaaggatc acatatcaaa
      721 ggggaagctc aggtggaggg cactgggttc ccagctgacg gccctgtgat gacaaattcc
      781 ctgacagccg aggcccacgg cctgttcaac ttcgtgaagg acgccggcgc caagctgtgg
      841 gatgccgtga caggccagca cgacaaggac gaccaggcca agaaggtgca ggaacacctg
      901 aacaagaccg gcatccccga cgccgacaaa gtgaacatcc agatcgccga cggcaaggcc
      961 acagtgacag gcgacggact gagccaggaa gccaaagaaa agatcctggt ggccgtgggc
     1021 aacatcagcg gaatcgccag cgtggacgat caagtgaaaa ccgccacccc tgccaccgcc
     1081 agccagttct acacagtgaa gtccggcgac accctgagcg ccatcagcaa acaggtgtac
     1141 ggcaacgcca acctgtacaa caagattttc gaggccaaca agcccatgct gaagtccccc
     1201 gacaaaatct accccggcca ggtgctgaga atccccgagg aaatgtactt cgctgactgg
     1261 tgcgtatcca agaagacttg ccccaatgat aagactattg tctccacatt taagtgggct
     1321 tttatcaccg acaacggcaa acgctaccgg tctaccgctc gcaccacata cacattcgcc
     1381 aagccaatgg ccgcaaatta cctcaaaaat cagccgatgt atgtctttcg gaaaacagag
     1441 ctgaaacatt ctaagaccga actgaatttt aaagagtggc aaaaggcctt tacggacgtg
     1501 atgggcatgg acgagctgta taagtaataa gaattcgaag cttggctgtt ttggtggatg
     1561 agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca
     1621 gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt
     1681 gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca
     1741 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     1801 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
     1861 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta
     1921 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt
     1981 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2041 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2101 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2161 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2221 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2281 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     2341 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2401 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2461 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2521 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2581 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     2641 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2701 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2761 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2821 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2881 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2941 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3001 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3061 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3121 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3181 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3241 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3301 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3361 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3421 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3481 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3541 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3601 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3661 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3721 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3781 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3841 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     3901 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3961 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     4021 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4081 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4141 cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca tgcataatgt gcctgtcaaa
     4201 tggacgaagc agggattctg caaaccctat gctactccgt caagccgtca attgtctgat
     4261 tcgttaccaa ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac
     4321 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat
     4381 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca
     4441 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct
     4501 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga
     4561 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat
     4621 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct
     4681 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga
     4741 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg
     4801 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt
     4861 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc
     4921 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca
     4981 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt
     5041 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg
     5101 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac
     5161 tcccgccatt cagag
//
