LOCUS       sequence-453948-        5411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chemigenetic indicator for potassium ion - low affinity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5411)
  AUTHORS   Cheng D, Ouyang Z, He X, Nasu Y, Wen Y, Terai T, Campbell RE
  TITLE     High-Performance Chemigenetic Potassium Ion Indicator.
  JOURNAL   J Am Chem Soc. 2024 Dec 25;146(51):35117-35128. doi: 
            10.1021/jacs.4c10917.
  PUBMED    39601449
REFERENCE   2  (bases 1 to 5411)
  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/jacs"; 
            journalName: "J Am Chem Soc"; date: "2024-12-25- 25"; volume: "146";
            issue: "51"; pages: "35117-35128"
COMMENT     Sequence Label: pBAD-HaloKbp1b
FEATURES             Location/Qualifiers
     source          1..5411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     186..205
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             309..326
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             330..362
                     /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(366..374,375..389)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1790..1807)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1960..2046
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2138..2165
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2184..2275
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2276..2344,2345..3136)
                     /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(2494..2513)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3307..3895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3796..3815
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4049..4066
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4081..4221
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4147..4166)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4307..4329
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4485..5363)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(5390..5411,1..263)
                     /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)"
ORIGIN
        1 catcagacat tgccgtcact gcgtctttta ctggctcttc tcgctaacca aaccggtaac
       61 cccgcttatt aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa
      121 caaaagtgtc tataatcacg gcagaaaagt ccacattgat tatttgcacg gcgtcacact
      181 ttgctatgcc atagcatttt tatccataag attagcggat cctacctgac gctttttatc
      241 gcaactctct actgtttctc catacccgtt ttttgggcta acaggaggaa ttaaccatgg
      301 ggggttctca tcatcatcat catcatggta tggctagcat gactggtgga cagcaaatgg
      361 gtcgggatct gtacgacgat gacgataagg atccgagctc gagaatggca gaaatcggta
      421 ctggctttcc attcgatccc cattatgtgg aagtcctggg cgagcgcatg cactacgtcg
      481 atgttggtcc gcgcgatggc acccctttgc tgttcctgca cggtaacccg acctcctcct
      541 acgtgtggcg caacatcatc ccgcatgttg caccgaccca tcgctgcatt gctccagacc
      601 tgatcggtat gggcaaatcc gacaaaccag acctgggtta tttcttcgac gaccacgtcc
      661 gcttcatgga tgccttcatc gaagccctgg gtctggaaga ggtcgtcctg gtcattcacg
      721 actggggctc cgctctgggt ttccactggg ccaagcgcaa tccagagcgc gtcaaaggta
      781 ttgcatttat ggagttcatc cgccctatcc cgacctggga cgaatggcca gaatttcttg
      841 tgggcccgtt caacttcgtg aaggacgccg gcgagaagct gtgggaagcc gtgacaggcc
      901 agcgcgacaa ggatgacctg gccaagaagg tgtgggaaca cctgcacaag accggcatcc
      961 ccgacgccga caaagtgaac atccaggtcg ccgacggcaa ggccacagtg gcaggcgacg
     1021 gactgagcca ggaagccaaa gaaaagatcc tggtggccgt gggcaacatc agcggaatcg
     1081 ccagcgtgga cgaccaagtg aaaaccgcca cccctgccac cgccagccag ttctacacag
     1141 tgaagtccgg cgacaccctg agcgccatca gcaatcaggt gtacggcaac gccgacctgt
     1201 acaacaagat tttcgtggcc aacaagccca tgctgaagtc ccccgacaag atctaccccg
     1261 gccaggcgct gagaatcccc tatagtcttg cccgcgagac cttccaggcc ttccgcacca
     1321 ccgacgtcgg ccgcaagctg atcatcgatc agaacgtttt tatcgagggt acgctgccga
     1381 tgggtgtcgt ccgcccgctg actgaagtcg agatggacca ttaccgcgag ccgttcctga
     1441 atcctgttga ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc gccggtgagc
     1501 cagcgaacat cgtcgcgctg gtcgaagaat acatgggctg gctgcaccag tcccctgtcc
     1561 cgaagctgct gttctggggc accccaggcg ttctgatccc accggccgaa gccgcccgcc
     1621 tggccaaaag cctgcctaac tgcaaggccg tggacatcgg cccgggtctg aatctgctgc
     1681 aagaagacaa cccggacctg atcggcagcg agatcgcgcg ctggctgtcg acgttggaaa
     1741 tttccggctg aaagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga
     1801 ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg
     1861 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     1921 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     1981 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     2041 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     2101 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc
     2161 ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc
     2221 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2281 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2341 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2401 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2461 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     2521 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2581 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2641 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2701 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2761 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2821 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2881 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2941 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3001 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3061 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3121 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3181 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3241 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3301 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aacaaccacc
     3361 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3421 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3481 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3541 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3601 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3661 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3721 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3781 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3841 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3901 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3961 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4021 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4081 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     4141 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     4201 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4261 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4321 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg
     4381 aaggcgaagc ggcatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc
     4441 ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg
     4501 gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg
     4561 cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg
     4621 gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc
     4681 tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac
     4741 ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga
     4801 tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg
     4861 ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc
     4921 gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc
     4981 ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta
     5041 agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga
     5101 gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc
     5161 ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga
     5221 ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg
     5281 gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg
     5341 ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat
     5401 tgtccatatt g
//
