LOCUS       sequence-351345-        5175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of green fluorescent potassium indicator 
            mNG-Ec-Kbp-D9N (KRaION1 mutant D9N) .
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-mNG-Ec-Kbp-D9N
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        12..296
                     /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     219..238
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             339..356
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1587..1604)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1757..1843
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1935..1962
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1981..2072
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2073..2141,2142..2933)
                     /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(2291..2310)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3104..3692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3593..3612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3846..3863
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3878..4018
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3944..3963)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4104..4126
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4282..5160)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt
       61 ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca
      121 aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa
      181 agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat
      241 aagattagcg gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc
      301 gttttttggg ctaacaggag gaattaacca tggggagcca ccatcaccat caccatggca
      361 gatctatggt ctcaaaggga gaggaggaaa acatggctag tctgcctgct acccacgaac
      421 tgcatatttt tggctccata aatggcatcg atttcgacat ggtaggccaa ggcaccggta
      481 atccaaacga cggctatgag gagctgaacc tgaagtccac catgggcgat ctgcagttca
      541 gtccctggat cctggtgccg cacattgggt acgggtttca ccagtatctt ccctatccag
      601 acgggatgtc accatttcag gctgcaatgg tcgacgggtc cggataccaa gtgcaccgca
      661 ctatgcagtt tgaggacggt gcctcactga ccgtgaatta cagatacacc tacgaaggat
      721 cacatatcaa aggggaagct caggtggagg gcactgggtt cccagctgac ggccctgtga
      781 tgacaaattc cctgacagcc gaggcccacg gcctgttcaa cttcgtgaag aacgccggcg
      841 agaagctgtg ggatgccgtg acaggccagc acgacaagga cgaccaggcc aagaaggtgc
      901 aggaacacct gaacaagacc ggcatccccg acgccgacaa agtgaacatc cagatcgccg
      961 acggcaaggc cacagtgaca ggcgacggac tgagccagga agccaaagaa aagatcctgg
     1021 tggccgtggg caacatcagc ggaatcgcca gcgtggacga tcaagtgaaa accgccaccc
     1081 ctgccaccgc cagccagttc tacacagtga agtccggcga caccctgagc gccatcagca
     1141 aacaggtgta cggcaacgcc aacctgtaca acaagatttt cgaggccaac aagcccatgc
     1201 tgaagtcccc cgacaaaatc taccccggcc aggtgctgag aatccccgag gaaatgtact
     1261 tcgctgactg gtgcgtatcc aagaagactt gccccaatga taagactatc gtctccacat
     1321 ttaagtgggc ttttatcacc gacaacggca aacgctaccg gtctaccgct cgcaccacat
     1381 acacattcgc caagccaatg gccgcaaatt acctcaaaaa tcagccgatg tatgtctttc
     1441 ggaaaacaga gctgaaacat tctaagaccg aactgaattt taaagagtgg caaaaggcct
     1501 ttacggacgt gatgggcatg gacgagctgt ataagtaata agaattcgaa gcttggctgt
     1561 tttggtggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1621 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     1681 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     1741 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1801 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     1861 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     1921 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     1981 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2041 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2101 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2161 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2221 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2281 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     2341 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2401 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2461 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2521 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2581 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2641 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2701 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2761 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2821 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2881 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2941 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3001 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3061 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3121 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3181 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3241 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3301 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3361 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3421 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3481 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3541 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3601 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3661 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3721 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3781 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3841 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     3901 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     3961 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     4021 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4081 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4141 accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcataatg
     4201 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
     4261 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
     4321 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
     4381 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
     4441 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
     4501 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
     4561 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
     4621 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
     4681 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
     4741 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
     4801 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
     4861 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
     4921 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
     4981 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     5041 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     5101 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat
     5161 acttttcata ctccc
//
