LOCUS       sequence-216972-        5231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of genetically encoded green fluorescent 
            potassium ion indicator GINKO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5231)
  AUTHORS   Shen Y, Wu SY, Rancic V, Aggarwal A, Qian Y, Miyashita SI, Ballanyi 
            K, Campbell RE, Dong M
  TITLE     Genetically encoded fluorescent indicators for imaging intracellular
            potassium ion concentration.
  JOURNAL   Commun Biol. 2019 Jan 14;2:18. doi: 10.1038/s42003-018-0269-2. 
            eCollection 2019.
  PUBMED    30652129
REFERENCE   2  (bases 1 to 5231)
  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: "Commun 
            Biol."; date: "2019-01-14"; volume: "2:18. doi"; pages: " 
            10.1038/s42003-018-0269-2. eCollection 2019"
COMMENT     Sequence Label: pBAD-GINKO1
FEATURES             Location/Qualifiers
     source          1..5231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             14..31
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             35..67
                     /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(71..79,80..94)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(155..176)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(416..435)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1210..1231
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1315..1332)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1485..1571
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1663..1690
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1709..1800
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1801..1869,1870..2661)
                     /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(2019..2038)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2832..3420
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3321..3340
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3574..3591
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3606..3746
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3672..3691)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3832..3854
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4010..4888)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4915..5199
                     /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     5122..5141
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 catggggggt tctcatcatc atcatcatca tggtatggct agcatgactg gtggacagca
       61 aatgggtcgg gatctgtacg acgatgacga taaggatccg agctcgagca tggtgagcaa
      121 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      181 cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg gcaagctgac
      241 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
      301 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
      361 cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct tcaaggacga
      421 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
      481 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
      541 caacctgccg gacggcctgt tcaacttcgt gaaggacgcc ggcgagaagc tgtgggatgc
      601 cgtgacaggc cagcacgaca aggacgacca ggccaagaag gtgcaggaac acctgaacaa
      661 gaccggcatc cccgacgccg acaaagtgaa catccagatc gccgacggca aggccacagt
      721 gacaggcgac ggactgagcc aggaagccaa agaaaagatc ctggtggccg tgggcaacat
      781 cagcggaatc gccagcgtgg acgatcaagt gaaaaccgcc acccctgcca ccgccagcca
      841 gttctacaca gtgaagtccg gcgacaccct gagcgccatc agcaaacagg tgtacggcaa
      901 cgccaacctg tacaacaaga ttttcgaggc caacaagccc atgctgaagt cccccgacaa
      961 aatctacccc ggccaggtgc tgagaatccc cgaggagctg gagaacgtct atatcaaggc
     1021 cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg
     1081 cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1141 gctgcccgac aaccactacc tgagcgtgca gtccaaactt tcgaaagacc ccaacgagaa
     1201 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1261 cgagctgtac aagtaaaagc ttggctgttt tggcggatga gagaagattt tcagcctgat
     1321 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
     1381 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
     1441 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     1501 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     1561 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     1621 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
     1681 atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg
     1741 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     1801 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1861 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     1921 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     1981 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     2041 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     2101 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     2161 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     2221 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     2281 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2341 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     2401 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2461 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     2521 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2581 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2641 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     2701 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     2761 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     2821 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaacaa
     2881 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2941 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     3001 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3061 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3121 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3181 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3241 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3301 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3361 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3421 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3481 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3541 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3601 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     3661 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     3721 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     3781 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3841 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc
     3901 gcgcgaaggc gaagcggcat gcataatgtg cctgtcaaat ggacgaagca gggattctgc
     3961 aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact
     4021 tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc
     4081 cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac
     4141 cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt
     4201 ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac
     4261 gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca
     4321 ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg
     4381 actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca
     4441 gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga
     4501 aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc
     4561 agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt
     4621 cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat
     4681 cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg
     4741 tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac
     4801 cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca
     4861 gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa
     4921 ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc
     4981 taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca
     5041 tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt
     5101 tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta gcggatccta
     5161 cctgacgctt tttatcgcaa ctctctactg tttctccata cccgtttttt gggctaacag
     5221 gaggaattaa c
//
