LOCUS       sequence-351347-        5175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of green fluorescent potassium indicator 
            mNG-C-Kbp.
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-C-Kbp
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        40..324
                     /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     247..266
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             367..384
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1615..1632)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1785..1871
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1963..1990
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2009..2100
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2101..2169,2170..2961)
                     /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(2319..2338)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3132..3720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3621..3640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3874..3891
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3906..4046
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3972..3991)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4132..4154
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(4310..5175,1..13))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat tgtccatatt
       61 gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc aaaccggtaa
      121 ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta
      181 acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac
      241 tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga cgctttttat
      301 cgcaactctc tactgtttct ccatacccgt tttttgggct aacaggagga attaaccatg
      361 gggagccacc atcaccatca ccatggcaga tctatggtct caaagggaga ggaggaaaac
      421 atggctagtc tgcctgctac ccacgaactg catatttttg gctccataaa tggcatcgat
      481 ttcgacatgg taggccaagg caccggtaat ccaaacgacg gctatgagga gctgaacctg
      541 aagtccacca tgggcgatct gcagttcagt ccctggatcc tggtgccgca cattgggtac
      601 gggtttcacc agtatcttcc ctatccagac gggatgtcac catttcaggc tgcaatggtc
      661 gacgggtccg gataccaagt gcaccgcact atgcagtttg aggacggtgc ctcactgacc
      721 gtgaattaca gatacaccta cgaaggatca catatcaaag gggaagctca ggtggagggc
      781 actgggttcc cagctgacgg ccctgtgatg acaaattccc tgacagccga ggcccacatg
      841 ggcctgttca acttcgtgaa ggaggccggc gagaagctgt gggacaccgt gaccggcaac
      901 gccaccgccg aggaccagag cgccaagctg aaggaccacc tgaacaagac cggcctgccc
      961 ggcatcgaca aggtggacgt gcaggtggtg gacggcaagg ccgtggtgac cggcgacgcc
     1021 gtgagccagg agctgaagga gaagatcctg gtggccgtgg gcaacgtggc cggcatcagc
     1081 ggcgtggagg acaaggtgag cgtggagcag gccgccgccg agagcaggtt ctacaccgtg
     1141 aagaagggcg acaccctgag cgccgtgagc aaggaggtgt acggcaacgc caacctgtac
     1201 aacaagatct tcgaggccaa caagcccatg ctgagcagcc ccgacaagat ctaccccggc
     1261 caggtgctga ggatccccga gatgtacttc gctgactggt gcgtatccaa gaagacttgc
     1321 cccaatgata agactatcgt ctccacattt aagtgggctt ttatcaccga caacggcaaa
     1381 cgctaccggt ctaccgctcg caccacatac acattcgcca agccaatggc cgcaaattac
     1441 ctcaaaaatc agccgatgta tgtctttcgg aaaacagagc tgaaacattc taagaccgaa
     1501 ctgaatttta aagagtggca aaaggccttt acggacgtga tgggcatgga cgagctgtat
     1561 aagtaataag aattcgaagc ttggctgttt tggtggatga gagaagattt tcagcctgat
     1621 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
     1681 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
     1741 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     1801 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     1861 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     1921 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
     1981 atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg
     2041 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2101 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     2161 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     2221 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     2281 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     2341 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     2401 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     2461 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     2521 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     2581 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2641 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     2701 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2761 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     2821 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2881 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2941 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     3001 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3061 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3121 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3181 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3241 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     3301 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3361 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3421 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3481 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3541 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3601 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3661 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3721 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3781 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3841 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3901 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     3961 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     4021 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     4081 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4141 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc
     4201 gcgcgaaggc gaagcggcat gcataatgtg cctgtcaaat ggacgaagca gggattctgc
     4261 aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact
     4321 tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc
     4381 cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac
     4441 cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt
     4501 ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac
     4561 gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca
     4621 ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg
     4681 actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca
     4741 gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga
     4801 aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc
     4861 agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt
     4921 cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat
     4981 cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg
     5041 tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac
     5101 cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca
     5161 gcaggggatc atttt
//
