LOCUS       sequence-351349-        5166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of green fluorescent potassium indicator 
            mNG-Pa-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-Pa-Kbp
FEATURES             Location/Qualifiers
     source          1..5166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        65..349
                     /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     272..291
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             392..409
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1631..1648)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1801..1887
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1979..2006
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2025..2116
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2117..2185,2186..2977)
                     /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(2335..2354)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3148..3736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3637..3656
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3890..3907
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3922..4062
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3988..4007)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4148..4170
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(4326..5166,1..38))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc
       61 agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg
      121 ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg
      181 accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac
      241 attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta
      301 gcggatccta cctgacgctt tttatcgcaa ctctctactg tttctccata cccgtttttt
      361 gggctaacag gaggaattaa ccatggggag ccaccatcac catcaccatg gcagatctat
      421 ggtctcaaag ggagaggagg aaaacatggc tagtctgcct gctacccacg aactgcatat
      481 ttttggctcc ataaatggca tcgatttcga catggtaggc caaggcaccg gtaatccaaa
      541 cgacggctat gaggagctga acctgaagtc caccatgggc gatctgcagt tcagtccctg
      601 gatcctggtg ccgcacattg ggtacgggtt tcaccagtat cttccctatc cagacgggat
      661 gtcaccattt caggctgcaa tggtcgacgg gtccggatac caagtgcacc gcactatgca
      721 gtttgaggac ggtgcctcac tgaccgtgaa ttacagatac acctacgaag gatcacatat
      781 caaaggggaa gctcaggtgg agggcactgg gttcccagct gacggccctg tgatgacaaa
      841 ttccctgaca gccgaggccc acatgggcat cttcgccttc gtgaaggagg ccggcgagaa
      901 gctgtgggac accctgaccg gccacgaggc ccaggccgcc gagagcctga aggagcacgt
      961 ggccaaggtg ggcctgggca accccaacat ccaggtgagc gtggagggcg acaaggtgat
     1021 cgccagcggc gaggtggcca gccaggagga gaaggagaag atcctgctgg ccctgggcaa
     1081 cgtggccggc gtgagcagcg tggaggacaa gatcaccgtg gcccaggccg cccccgaggc
     1141 caggttcgtg accgtgaaga agggcgacac cctgagcgcc atcgccaagg ccgagtacgg
     1201 caacgccaac gcctacatga agatcttcga ggccaacaag cccatgctga gccaccccga
     1261 caagatctac cccggccagg tgctgaggat ccccgagatg tacttcgctg actggtgcgt
     1321 atccaagaag acttgcccca atgataagac tatcgtctcc acatttaagt gggcttttat
     1381 caccgacaac ggcaaacgct accggtctac cgctcgcacc acatacacat tcgccaagcc
     1441 aatggccgca aattacctca aaaatcagcc gatgtatgtc tttcggaaaa cagagctgaa
     1501 acattctaag accgaactga attttaaaga gtggcaaaag gcctttacgg acgtgatggg
     1561 catggacgag ctgtataagt aataagaatt cgaagcttgg ctgttttggt ggatgagaga
     1621 agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt
     1681 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
     1741 gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat
     1801 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     1861 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
     1921 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
     1981 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt tatttttcta
     2041 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2101 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2161 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2221 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2281 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2341 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2401 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2461 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2521 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2581 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2641 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     2701 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2761 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2821 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2881 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2941 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3001 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3061 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3121 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3181 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3241 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     3301 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3361 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3421 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3481 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3541 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3601 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3661 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3721 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3781 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3841 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3901 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3961 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4021 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     4081 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4141 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     4201 cagcagatca attcgcgcgc gaaggcgaag cggcatgcat aatgtgcctg tcaaatggac
     4261 gaagcaggga ttctgcaaac cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt
     4321 accaattatg acaacttgac ggctacatca ttcacttttt cttcacaacc ggcacggaac
     4381 tcgctcgggc tggccccggt gcatttttta aatacccgcg agaaatagag ttgatcgtca
     4441 aaaccaacat tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa aagcagcttc
     4501 gcctggctga tacgttggtc ctcgcgccag cttaagacgc taatccctaa ctgctggcgg
     4561 aaaagatgtg acagacgcga cggcgacaag caaacatgct gtgcgacgct ggcgatatca
     4621 aaattgctgt ctgccaggtg atcgctgatg tactgacaag cctcgcgtac ccgattatcc
     4681 atcggtggat ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc
     4741 agatttatcg ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc
     4801 ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg
     4861 gcaaatattg acggccagtt aagccattca tgccagtagg cgcgcggacg aaagtaaacc
     4921 cactggtgat accattcgcg agcctccgga tgacgaccgt agtgatgaat ctctcctggc
     4981 gggaacagca aaatatcacc cggtcggcaa acaaattctc gtccctgatt tttcaccacc
     5041 ccctgaccgc gaatggtgag attgagaata taacctttca ttcccagcgg tcggtcgata
     5101 aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag atgggcatta
     5161 aacgag
//
