LOCUS       sequence-351350-        5166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of green fluorescent potassium indicator 
            mNG-Hv-Kbp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5166)
  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 5166)
  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-Hv-Kbp
FEATURES             Location/Qualifiers
     source          1..5166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..290
                     /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     213..232
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             333..350
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1572..1589)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1742..1828
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1920..1947
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1966..2057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2058..2126,2127..2918)
                     /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(2276..2295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3089..3677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3578..3597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3831..3848
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3863..4003
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3929..3948)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4089..4111
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4267..5145)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg
       61 gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg
      121 gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca
      181 cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt
      241 agcggatcct acctgacgct ttttatcgca actctctact gtttctccat acccgttttt
      301 tgggctaaca ggaggaatta accatgggga gccaccatca ccatcaccat ggcagatcta
      361 tggtctcaaa gggagaggag gaaaacatgg ctagtctgcc tgctacccac gaactgcata
      421 tttttggctc cataaatggc atcgatttcg acatggtagg ccaaggcacc ggtaatccaa
      481 acgacggcta tgaggagctg aacctgaagt ccaccatggg cgatctgcag ttcagtccct
      541 ggatcctggt gccgcacatt gggtacgggt ttcaccagta tcttccctat ccagacggga
      601 tgtcaccatt tcaggctgca atggtcgacg ggtccggata ccaagtgcac cgcactatgc
      661 agtttgagga cggtgcctca ctgaccgtga attacagata cacctacgaa ggatcacata
      721 tcaaagggga agctcaggtg gagggcactg ggttcccagc tgacggccct gtgatgacaa
      781 attccctgac agccgaggcc cacatgggca tctgggactt cgtgaagagc gccggcagga
      841 agatcggcat cggcagcgag gaggccgccc ccgccgccga cgacctgaag aaggagctgg
      901 acagctacaa gctgggcacc gacgacgtga aggtggaggt ggtgggcgac aaggccgtga
      961 tcaccggcaa cgtggccgac cagagcatcc tggagaaggc catcgtggcc gtgggcaaca
     1021 ccctgggcat cagcaaggtg gagagcaacg tggaggtgcc cgacgccaag gagcccgtgt
     1081 tccacaccgt gaagaagggc gacaccctgt gggccatcgc cagcaagcag tacggcaacg
     1141 gcagcaagta caacgccatc ttcgaggcca acaagcccat gctgagccac cccgacaaga
     1201 tctaccccgg ccaggtgctg aggatccccg acctgggcat gtacttcgct gactggtgcg
     1261 tatccaagaa gacttgcccc aatgataaga ctatcgtctc cacatttaag tgggctttta
     1321 tcaccgacaa cggcaaacgc taccggtcta ccgctcgcac cacatacaca ttcgccaagc
     1381 caatggccgc aaattacctc aaaaatcagc cgatgtatgt ctttcggaaa acagagctga
     1441 aacattctaa gaccgaactg aattttaaag agtggcaaaa ggcctttacg gacgtgatgg
     1501 gcatggacga gctgtataag taataagaat tcgaagcttg gctgttttgg tggatgagag
     1561 aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat
     1621 ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa
     1681 cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     1741 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1801 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     1861 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     1921 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
     1981 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2041 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2101 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2161 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2221 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2281 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     2341 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2401 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2461 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2521 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2581 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2641 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2701 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2761 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2821 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2881 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2941 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3001 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3061 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3121 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3181 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3241 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3301 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3361 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3421 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3481 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3541 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3601 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3661 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3721 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3781 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3841 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3901 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3961 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     4021 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4081 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4141 gcagcagatc aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct gtcaaatgga
     4201 cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt
     4261 taccaattat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa
     4321 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc
     4381 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt
     4441 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg
     4501 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc
     4561 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc
     4621 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag
     4681 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg
     4741 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt
     4801 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac
     4861 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg
     4921 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac
     4981 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat
     5041 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt
     5101 aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc
     5161 gccatt
//
