LOCUS       sequence-455857-        5411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chemigenetic indicator for potassium ion - medium 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-HaloKbp1c
FEATURES             Location/Qualifiers
     source          1..5411
                     /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             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /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(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1812..1829)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1982..2068
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2160..2187
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2206..2297
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2298..2366,2367..3158)
                     /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(2516..2535)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3329..3917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3818..3837
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4071..4088
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4103..4243
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4169..4188)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4329..4351
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4507..5385)
                     /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 ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgagaatgg cagaaatcgg tactggcttt ccattcgatc cccattatgt ggaagtcctg
      481 ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcaccccttt gctgttcctg
      541 cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc
      601 catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc agacctgggt
      661 tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct gggtctggaa
      721 gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg ggccaagcgc
      781 aatccagagc gcgtcaaagg tattgcattt atggagttca tccgccctat cccgacctgg
      841 gacgaatggc cagaatttct tgtgggcccg ttcaacttcg tggaggacgc cggcgtgaag
      901 ctgtgggaag ccgtgacagg ccagcacgac aaggatgacc tggccaagaa ggtgtgggaa
      961 cacctgcaca agaccggcat ccccgacgcc gacaaagtga acatccaggt cgccgacggc
     1021 aaggccacag tggcaggcgg cggactgagc caggatgcca aagaaaagat cctggtggcc
     1081 gtgggcaaca tcagcggaat cgccagcgtg gacgaccatg tgaaaaccgc cacccctgcc
     1141 accgccagcc agttctacac agtgaagtcc ggcgacaccc tgagcgccat cagcaatcag
     1201 gtgtacggca acgccgacct gtacaacaag attttcgagg ccaacaagcc catgctgaag
     1261 tcccccgacg agatctaccc cggccaggcg ctgagaatcc cctatagtct tgcccgcgag
     1321 accttccagg ccttccgctc caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt
     1381 tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac
     1441 cattaccgcg agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac
     1501 gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac
     1561 tggctgcacc agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc
     1621 ccaccggccg aagccgcccg cctggccaaa agcctgccta actgcaaggc cgtggacatc
     1681 ggcccgggtc tgaatctgct gcaagaagac aacccggtcc tgatcggcag cgagatcgcg
     1741 cgctggctgt cgacgttgga aatttccggc tgaaagcttg gctgttttgg cggatgagag
     1801 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     1861 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     1921 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     1981 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     2041 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     2101 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     2161 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
     2221 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2281 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2341 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2401 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2461 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2521 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     2581 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2641 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2701 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2761 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2821 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2881 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2941 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3001 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3061 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3121 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3181 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3241 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3301 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3361 gcttgcaaac aaaacaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3421 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3481 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3541 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3601 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3661 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3721 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3781 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3841 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3901 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3961 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4021 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4081 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4141 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4201 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     4261 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4321 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4381 gcagcagatc aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct gtcaaatgga
     4441 cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt
     4501 taccaattat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa
     4561 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc
     4621 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt
     4681 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg
     4741 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc
     4801 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc
     4861 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag
     4921 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg
     4981 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt
     5041 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac
     5101 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg
     5161 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac
     5221 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat
     5281 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt
     5341 aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc
     5401 gccattcaga g
//
