LOCUS       sequence-348390-        5230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of genetically encoded green fluorescent 
            potassium ion indicator GINKO2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5230)
  AUTHORS   Wu SY, Wen Y, Serre NBC, Laursen CCH, Dietz AG, Taylor BR, Drobizhev
            M, Molina RS, Aggarwal A, Rancic V, Becker M, Ballanyi K, Podgorski 
            K, Hirase H, Nedergaard M, Fendrych M, Lemieux MJ, Eberl DF, Kay AR,
            Campbell RE, Shen Y
  TITLE     A sensitive and specific genetically-encoded potassium ion biosensor
            for in vivo applications across the tree of life.
  JOURNAL   PLoS Biol. 2022 Sep 6;20(9):e3001772. doi: 
            10.1371/journal.pbio.3001772. eCollection 2022 Sep.
  PUBMED    36067248
REFERENCE   2  (bases 1 to 5230)
  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"; journalName: "PLoS 
            Biol."; date: "2022-09-6"; volume: "20(9):e3001772. doi"; pages: " 
            10.1371/journal.pbio.3001772. eCollection 2022 Sep"
COMMENT     Sequence Label: pBAD-GINKO2
FEATURES             Location/Qualifiers
     source          1..5230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        104..388
                     /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     311..330
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             434..451
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             455..487
                     /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(491..499,500..514)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(575..596)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(836..855)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1630..1651
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1734..1751)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1904..1990
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2082..2109
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2128..2219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2220..2288,2289..3080)
                     /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(2438..2457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3251..3839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3740..3759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3993..4010
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4025..4165
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4091..4110)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4251..4273
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(4429..5230,1..77))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca
       61 ttttgcgctt cagccatact tttcatactc ccgccattca gagaagaaac caattgtcca
      121 tattgcatca gacattgccg tcactgcgtc ttttactggc tcttctcgct aaccaaaccg
      181 gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat gacaaaaacg
      241 cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt gcacggcgtc
      301 acactttgct atgccatagc atttttatcc ataagattag cggatcctac ctgacgcttt
      361 ttatcgcaac tctctactgt ttctccatac ccgttttttg ggctaacagg aggaattaac
      421 catggggggt tctcatcatc atcatcatca tggtatggct agcatgactg gtggacagca
      481 aatgggtcgg gatctgtacg acgatgacga taaggatccg agctcgagca tggtgagcaa
      541 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      601 cggccacaag ttcagcgtgt acggcgaggg tgagggcgat gccacctacg gcaagctgac
      661 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccactc tcgtgaccac
      721 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
      781 cttcaagtcc gccatgcccg aaggctacac ccaggagcgc accatcttct tcgaggacga
      841 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
      901 cgagctggag ggcatcgact tcaaggagga cggcaacatt ctggggcaca agctggagta
      961 caacctgccg gacggcctgt tcaacttcgt gaaggacgcc ggcgagaagc tgtgggatgc
     1021 cgtgacaggc cagcacgaca aggacgacca ggccaagaag gtgtgggaac acctgcacaa
     1081 gaccggcatc cccgacgccg acaaagtgaa catccaggtc gccgacggca aggccacagt
     1141 ggcaggcgac ggactgagcc aggaagccaa agaaaagatc ctggtggccg tgggcaacat
     1201 cagcggaatc gccagcgtgg acgaccaagt gaaaaccgcc acccctgcca ccgccagcca
     1261 gttctacaca gtgaagtccg gcgacaccct gagcgccatc agcaatcagg tgtacggcaa
     1321 cgccgacctg tacaacaaga ttttcgaggc caacaagccc atgctgaagt cccccgacaa
     1381 aatctacccc ggccaggtgc tgagaatccc ctatgctgct aataacgtct atatcaaggc
     1441 cgacaaggag aagaacggca tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg
     1501 cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1561 gctgcccgac aaccactacc tgagcgtgca gtccagactt tctaaagacc ccaacgagaa
     1621 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1681 cgagctgtac atgtaaagct tggctgtttt ggcggatgag agaagatttt cagcctgata
     1741 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc
     1801 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt
     1861 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
     1921 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     1981 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     2041 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
     2101 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt
     2161 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2221 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2281 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2341 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2401 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2461 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2521 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2581 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2641 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2701 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2761 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2821 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2881 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2941 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3001 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3061 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3121 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3181 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3241 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3301 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3361 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3421 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3481 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3541 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3601 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3661 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3721 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3781 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3841 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3901 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3961 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4021 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     4081 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     4141 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     4201 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4261 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg
     4321 cgcgaaggcg aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca
     4381 aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt
     4441 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
     4501 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
     4561 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
     4621 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
     4681 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
     4741 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
     4801 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
     4861 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
     4921 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
     4981 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
     5041 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
     5101 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
     5161 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
     5221 gttggcctca
//
