LOCUS       sequence-194090-        6972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bongwoori with the CC1-Pos6 linker length and composition.(* super 
            ecliptic pHluorin A227D).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6972)
  AUTHORS   Lee S, Geiller T, Jung A, Nakajima R, Song YK, Baker BJ
  TITLE     Improving a genetically encoded voltage indicator by modifying the 
            cytoplasmic charge composition.
  JOURNAL   Sci Rep. 2017 Aug 15;7(1):8286. doi: 10.1038/s41598-017-08731-2.
  PUBMED    28811673
REFERENCE   2  (bases 1 to 6972)
  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: "Sci Rep."; 
            date: "2017-08-15"; volume: "7(1):8286. doi"; pages: " 
            10.1038/s41598-017-08731-2"
COMMENT     Sequence Label: Bongwoori-Pos6
FEATURES             Location/Qualifiers
     source          1..6972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..199
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(208..636)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(318..339)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     530..549
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    682..906
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     895..912
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(957..1673)
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLE
                     FVTADGITHGMDELYK"
     primer_bind     complement(1000..1020)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     1615..1643
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     complement(2424..2443)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2425..2443)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2488..2691)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2517..2537)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2692..3071
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3243..3262
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3337..3441
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3442..3510,3511..4302)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3660..3679)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4473..5061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4962..4981
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5215..5232
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5349..5370
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5385..5390,5391..5408,5409..5415)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5423..5439
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5428..5450
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5447..5463
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5447..5463
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5512..5633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5523..5542)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5577..5596
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5763..6788)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(5763..6788)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(join(6837..6972,1..194))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6851..6972,1..14)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6905..6924)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
       61 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
      121 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
      181 cctaactgac acacattcca cagaattaat tcgcgttaaa tttttgttaa atcagctcat
      241 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      301 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      361 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      421 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      481 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      541 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      601 cacccgccgc gcttaatgcg ccgctacagg gcgcgtgggg atacccccta gagccccagc
      661 tggttctttc cgcctcagaa gccatagagc ccaccgcatc cccagcatgc ctgctattgt
      721 cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc
      781 tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac
      841 cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag
      901 gcacagtcga ggctgatcag cgggtttaaa cgggccctct agactcgagt tctagatcat
      961 ttgtatagtt catccatgcc atgtgtaatc ccatcagctg ttacaaactc aagaaggacc
     1021 atgtggtctc tcttttcgtt gggatctttc gaaagagtag aagttgtaaa caggtaatgg
     1081 ttgtctggta aaaggacggg cccatcgcca attggagtat tttgttgata atggtctgct
     1141 agttgaacgc ctccatcttc aatgttgtgt ctaattttga agttagcttt gattccattc
     1201 ttttgtttgt ctgccatgat gtacacctgg tgatcgttat agttgtattc caatttgtgt
     1261 ccaagaatgt ttccatcttc tttaaaatca atacctttta actcgattct attaacaagg
     1321 gtatcacctt caaacttgac ttcagcacgt gtcttgtagt tcccgtcatc tttgaaaaat
     1381 atagttcttt cctgtacata accttcgggc atggcactct tgaaaaagtc atgccgtttc
     1441 atatgatctg ggtatcttga aaagcattga acaccataag ttaaagtagt gacaagtgtt
     1501 ggccatggaa caggtagttt tccagtagtg caaataaatt taagggtaag ttttccgtat
     1561 gttgcatcac cttcaccctc tccactgaca gaaaatttgt gcccattaac atcaccatct
     1621 aattcaacaa gaattgggac aactccagtg aaaagttctt ctcctttact cataggcctc
     1681 cccatccttc tgtgggatct aaatattctt gctaagataa ccacacgcag caatctggcc
     1741 aacacaacca actgccccaa accatcggct cctgtttcgg taccgtattc atctaacaca
     1801 gtgtaaaata tcgtgacgac gaatgtgaca acgataatca aaccatcagc cacctcccaa
     1861 gggttggtga agaaaagctt gggcccgtag gcaaatatcc ttaatcctaa atccagcatg
     1921 aaataacaag aaagagccaa gtccatcccg tcataaaagg attgtgaaga ttcactttta
     1981 tcgataagac tgagatcaat gatcatgagg atgatgtcca agaaaattag gaagactcca
     2041 aagactcgca tccctagatg atcaatcact gctcgaatgc gaaactggac gcgacctaca
     2101 ccagtagtag gtatctccat tctcccatcg tccactccat gttcattctc gccccaccat
     2161 agacctgaaa tctcgggtat atctattcgt tcctcctgtc ttgtctctgg tttttcactc
     2221 ggtccttggt ctacatcatt ccaatgaact ttctttaccg attcactttt tcttggcgca
     2281 gctttcggcg gagcagtgac gtcaccattt atcgtgacgt caacgacgtt ccgcatgact
     2341 gcaccgtcaa cgccaactaa atcagctgga ggactaaaat ctgaaccgtc gaatccctcc
     2401 atggtgctag ccagcttggg tctccctata gtgagtcgta ttaatttcga taagccagta
     2461 agcagtgggt tctctagtta gccagagagc tctgcttata tagacctccc accgtacacg
     2521 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
     2581 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     2641 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
     2701 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     2761 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     2821 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     2881 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     2941 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     3001 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa
     3061 taatcaatgt caacgcgtat atctggcccg tacatcgcga agcagcgcaa aacgcctaac
     3121 cctaagcaga ttcttcatgc aattgtcggt caagccttgc cttgttgtag cttaaatttt
     3181 gctcgcgcac tactcagcga cctccaacac acaagcaggg agcagatact ggcttaacta
     3241 tgcggcatca gagcagattg tactgagagt gcaccatagg ggatcgggag atctcccgat
     3301 ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3361 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3421 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3481 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3541 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3601 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3661 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3721 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3781 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3841 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3901 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3961 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4021 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4081 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4141 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4201 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4261 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4321 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4381 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4441 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4501 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4561 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4621 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4681 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4741 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4801 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4861 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4921 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4981 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5041 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5101 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5161 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5221 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     5281 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     5341 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     5401 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     5461 gaccatgatt acgccaagct ctagctagag gtcgacggta tacagacatg ataagataca
     5521 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     5581 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttggggtgg
     5641 gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa
     5701 acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtagcacg
     5761 tgctattcct ttgccctcgg acgagtgctg gggcgtcggt ttccactatc ggcgagtact
     5821 tctacacagc catcggtcca gacggccgcg cttctgcggg cgatttgtgt acgcccgaca
     5881 gtcccggctc cggatcggac gattgcgtcg catcgaccct gcgcccaagc tgcatcatcg
     5941 aaattgccgt caaccaagct ctgatagagt tggtcaagac caatgcggag catatacgcc
     6001 cggagccgcg gcgatcctgc aagctccgga tgcctccgct cgaagtagcg cgtctgctgc
     6061 tccatacaag ccaaccacgg cctccagaag aagatgttgg cgacctcgta ttgggaatcc
     6121 ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc ggccattgtc cgtcaggaca
     6181 ttgttggagc cgaaatccgc gtgcacgagg tgccggactt cggggcagtc ctcggcccaa
     6241 agcatcagct catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc catcacagtt
     6301 tgccagtgat acacatgggg atcagcaatc gcgcatatga aatcacgcca tgtagtgtat
     6361 tgaccgattc cttgcggtcc gaatgggccg aacccgctcg tctggctaag atcggccgca
     6421 gcgatcgcat ccatggcctc cgcgaccggc tgcagaacag cgggcagttc ggtttcaggc
     6481 aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc aataggtcag gctctcgctg
     6541 aattccccaa tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa gtgccgataa
     6601 acataacgat ctttgtagaa accatcggcg cagctattta cccgcaggac atatccacgc
     6661 cctcctacat cgaagctgaa agcacgagat tcttcgccct ccgagagctg catcaggtcg
     6721 gagacgctgt cgaacttttc gatcagaaac ttctcgacag acgtcgcggt gagttcaggc
     6781 tttttcatca cgtgctgatc agatccgaaa atggatatac aagctcccgg gagctttttg
     6841 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggcagag
     6901 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     6961 gaactgggcg ga
//
