LOCUS       sequence-194603-        6972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CC1-Pos6 is a modified version of CC1-M240 that has 4 arginines in 
            its linker. (* 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: CC1-Pos6
FEATURES             Location/Qualifiers
     source          1..6972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(51..71)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        56..385
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      236..371
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     298..317
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             434..1459
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             434..1459
                     /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"
     polyA_signal    1589..1710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1626..1645)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1680..1699
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1759..1775)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1759..1775)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1772..1794)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1783..1799
                     /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)."
     promoter        complement(join(1807..1813,1814..1831,1832..1837))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1852..1873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1990..2007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2161..2749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2241..2260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2920..3711,3712..3780))
                     /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     3543..3562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3781..3885)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3960..3979)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4151..4530
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4531..4734
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4685..4705
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4779..4798
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4779..4797
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5549..6265
                     /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(5579..5607)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6202..6222
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6310..6327)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6316..6540
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6586..6972,1..42)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6673..6692)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6883..6904
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
       61 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
      121 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
      181 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
      241 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
      301 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
      361 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
      421 tgatcagcac gtgatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
      481 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
      541 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
      601 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
      661 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
      721 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
      781 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
      841 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
      901 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
      961 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     1021 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     1081 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     1141 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     1201 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     1261 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     1321 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     1381 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     1441 tccgagggca aaggaatagc acgtgctacg agatttcgat tccaccgccg ccttctatga
     1501 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     1561 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     1621 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1681 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     1741 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1801 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1861 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1921 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1981 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2041 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2101 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2161 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2221 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2281 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2341 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2401 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2461 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2521 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2581 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2641 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2701 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2761 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2821 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2881 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2941 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3001 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3061 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3121 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3181 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3241 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3301 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3361 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3421 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3481 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3541 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3601 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3661 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3721 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3781 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3841 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3901 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     3961 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     4021 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     4081 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     4141 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     4201 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     4261 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     4321 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     4381 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     4441 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     4501 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     4561 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     4621 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     4681 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
     4741 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     4801 acccaagctg gctagcacca tggagggatt cgacggttca gattttagtc ctccagctga
     4861 tttagttggc gttgacggtg cagtcatgcg gaacgtcgtt gacgtcacga taaatggtga
     4921 cgtcactgct ccgccgaaag ctgcgccaag aaaaagtgaa tcggtaaaga aagttcattg
     4981 gaatgatgta gaccaaggac cgagtgaaaa accagagaca agacaggagg aacgaataga
     5041 tatacccgag atttcaggtc tatggtgggg cgagaatgaa catggagtgg acgatgggag
     5101 aatggagata cctactactg gtgtaggtcg cgtccagttt cggatccgag cagtgattga
     5161 tcatctaggg atgcgagtct ttggagtctt cctaattttc ttggacatca tcctcatgat
     5221 cattgatctc agtcttatcg ataaaagtga atcttcacaa tccttttatg acgggatggc
     5281 tttggctctt tcttgttatt tcatgctgga tttaggatta aggatatttg cctacgggcc
     5341 caagcttttc ttcaccaacc cttgggaggt ggctgatggt ttgattatcg ttgtcacatt
     5401 cgtcgtcacg atattttaca ctgtgttaga tgaatacggt accgaaacag gagccgatgg
     5461 tttggggcgg ttggttgtgt tggcccgttt gctgcgtgtg gttcggttag caagaatatt
     5521 tagatcccac agaaggatgg ggaggcctat gagtaaagga gaagaacttt tcactggagt
     5581 tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg
     5641 agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg
     5701 aaaactacct gttccatggc caacacttgt cactacttta acttatggtg ttcaatgctt
     5761 ttcaagatac ccagatcata tgaaacggca tgactttttc aagagtgcca tgcccgaagg
     5821 ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga
     5881 agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa
     5941 agaagatgga aacattcttg gacacaaatt ggaatacaac tataacgatc accaggtgta
     6001 catcatggca gacaaacaaa agaatggaat caaagctaac ttcaaaatta gacacaacat
     6061 tgaagatgga ggcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg
     6121 gcccgtcctt ttaccagaca accattacct gtttacaact tctactcttt cgaaagatcc
     6181 caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgatg ggattacaca
     6241 tggcatggat gaactataca aatgatctag aactcgagtc tagagggccc gtttaaaccc
     6301 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     6361 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     6421 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     6481 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     6541 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     6601 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     6661 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     6721 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     6781 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     6841 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     6901 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     6961 tttcggccta tt
//
