LOCUS       sequence-453947-        5411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chemigenetic indicator for potassium ion - high 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-HaloKbp1a
FEATURES             Location/Qualifiers
     source          1..5411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     200..219
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             323..340
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             344..376
                     /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(380..388,389..403)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1804..1821)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1974..2060
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2152..2179
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2198..2289
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2290..2358,2359..3150)
                     /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(2508..2527)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3321..3909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3810..3829
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4063..4080
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4095..4235
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4161..4180)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4321..4343
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4499..5377)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(5404..5411,1..277)
                     /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 aattgtccat attgcatcag acattgccgt cactgcgtct tttactggct cttctcgcta
       61 accaaaccgg taaccccgct tattaaaagc attctgtaac aaagcgggac caaagccatg
      121 acaaaaacgc gtaacaaaag tgtctataat cacggcagaa aagtccacat tgattatttg
      181 cacggcgtca cactttgcta tgccatagca tttttatcca taagattagc ggatcctacc
      241 tgacgctttt tatcgcaact ctctactgtt tctccatacc cgttttttgg gctaacagga
      301 ggaattaacc atggggggtt ctcatcatca tcatcatcat ggtatggcta gcatgactgg
      361 tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatccga gctcgagaat
      421 ggcagaaatc ggtactggct ttccattcga tccccattat gtggaagtcc tgggcgagcg
      481 catgcactac gtcgatgttg gtccgcgcga tggcacccct ttgctgttcc tgcacggtaa
      541 cccgacctcc tcctacgtgt ggcgcaacat catcccgcat gttgcaccga cccatcgctg
      601 cattgctcca gacctgatcg gtatgggcaa atccgacaaa ccagacctgg gttatttctt
      661 cgacgaccac gtccgcttca tggatgcctt catcgaagcc ctgggtctgg aagaggtcgt
      721 cctggtcatt cacgactggg gctccgctct gggtttccac tgggccaagc gcaatccaga
      781 gcgcgtcaaa ggtattgcat ttatggagtt catccgccct atcccgacct gggacgaatg
      841 gccagaattt cttgtgggcc cgttcaactt cgtgaaggac gccggcgaga agctgtggga
      901 agccgtgaca ggccagcacg acaaggatga cctggccaag aaggtgtggg aacacctgca
      961 caagaccggc atccccgacg ccgacaaagt gaacatccag gtcgccgacg gcaaggccac
     1021 agtggcaggc gacggactga gccaggaagc caaagaaaag atcctggtgg ccgtgggcaa
     1081 catcagcgga atcgccagcg tggacgacca agtgaaaacc gccacccctg ccaccgccag
     1141 ccagttctac acagtgaagt ccggcgacac cctgagcgcc atcagcaatc aggtgtacgg
     1201 caacgccgac ctgtacaaca agattttcga ggccaacaag cccatgctga agtcccccga
     1261 caagatctac cccggccagg cgctgagaat cccctatagt cttgcccgcg agaccttcca
     1321 ggccttccgc accaccgacg tcggccgcaa gctgatcatc gatcagaacg tttttatcga
     1381 gggtacgctg ccgatgggtg tcgtccgccc gctgactgaa gtcgagatgg accattaccg
     1441 cgagccgttc ctgaatcctg ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc
     1501 aatcgccggt gagccagcga acatcgtcgc gctggtcgaa gaatacatgg actggctgca
     1561 ccagtcccct gtcccgaagc tgctgttctg gggcacccca ggcgttctga tcccaccggc
     1621 cgaagccgcc cgcctggcca aaagcctgcc taactgcaag gccgtggaca tcggcccggg
     1681 tctgaatctg ctgcaagaag acaacccgga cctgatcggc agcgagatcg cgcgctggct
     1741 gtcgacgttg gaaatttccg gctgaaagct tggctgtttt ggcggatgag agaagatttt
     1801 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
     1861 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     1921 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     1981 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     2041 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     2101 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
     2161 tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat
     2221 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2281 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2341 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2401 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2461 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2521 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2581 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2641 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2701 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2761 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2821 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2881 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2941 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3001 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3061 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3121 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3181 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3241 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3301 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3361 acaaaacaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3421 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3481 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3541 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3601 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3661 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3721 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3781 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3841 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3901 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3961 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4021 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4081 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4141 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     4201 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     4261 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4321 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga
     4381 tcaattcgcg cgcgaaggcg aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag
     4441 ggattctgca aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt
     4501 atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg
     4561 ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa
     4621 cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc
     4681 tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat
     4741 gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc
     4801 tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg
     4861 gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta
     4921 tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca
     4981 ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata
     5041 ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt
     5101 gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca
     5161 gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac
     5221 cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat
     5281 cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt
     5341 atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca
     5401 gagaagaaac c
//
