LOCUS       sequence-46726-e        7167 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7167)
  AUTHORS   Jin L, Han Z, Platisa J, Wooltorton JR, Cohen LB, Pieribone VA
  TITLE     Single action potentials and subthreshold electrical events imaged 
            in neurons with a fluorescent protein voltage probe.
  JOURNAL   Neuron. 2012 Sep 6;75(5):779-85.
  PUBMED    22958819
REFERENCE   2  (bases 1 to 7167)
  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: "Neuron."; 
            date: "2012-09-6"; volume: "75(5)"; pages: "779-85"
COMMENT     Sequence Label: ArcLight-S249
FEATURES             Location/Qualifiers
     source          1..7167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             820..1536
                     /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(850..878)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1473..1493
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    1566..1700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1805..1825)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1810..2139
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1990..2125
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2052..2071
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2188..3213
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2188..3213
                     /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    3343..3464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3502..3518)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3502..3518)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3515..3537)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3526..3542
                     /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(3550..3556,3557..3574,3575..3580))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3595..3616
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3733..3750)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3904..4492)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3984..4003)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4663..5454,5455..5523))
                     /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     5286..5305
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5524..5628)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5654..6109)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5791..5812)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6003..6022
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6183..7167
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagctag cttgatttag gtgacactat agaatacaag
       61 cttatggagg gattcgacgg ttcagatttt agtcctccag ctgatttagt tggcgttgac
      121 ggtgcagtca tgcggaacgt cgttgacgtc acgataaatg gtgacgtcac tgctccgccg
      181 aaagctgcgc caagaaaaag tgaatcggta aagaaagttc attggaatga tgtagaccaa
      241 ggaccgagtg aaaaaccaga gacaagacag gaggaacgaa tagatatacc cgagatttca
      301 ggtctatggt ggggcgagaa tgaacatgga gtggacgatg ggagaatgga gatacctact
      361 actggtgtag gtcgcgtcca gtttcgtgtc cgagcagtga ttgatcatct agggatgcga
      421 gtctttggag tcttcctaat tttcttggac atcatcctca tgatcattga tctcagtctt
      481 ccaggaaaaa gtgaatcttc acaatccttt tatgacggga tggctttggc tctttcttgt
      541 tatttcatgc tggatttagg attaaggata tttgcctacg ggcccaagaa tttcttcacc
      601 aacccctggg aggttgctga tggtttgatt atcgttgtca cattcgtcgt cacgatattt
      661 tacactgtgt tagatgaata cgtgcaagaa acaggagccg atggtttggg gcaattggtt
      721 gtgttggccc gtttgctgcg tgtggttcga ttagcaagaa tattttattc ccaccaacaa
      781 atgaaggctt caagcagaag aacaatatca ggggatccca tgagtaaagg agaagaactt
      841 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
      901 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
      961 tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactacttt aacttatggt
     1021 gttcaatgct tttcaagata cccagatcat atgaaacggc atgacttttt caagagtgcc
     1081 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     1141 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     1201 attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataacgat
     1261 caccaggtgt acatcatggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     1321 agacacaaca ttgaagatgg aggcgttcaa ctagcagacc attatcaaca aaatactcca
     1381 attggcgatg ggcccgtcct tttaccagac aaccattacc tgtttacaac ttctactctt
     1441 tcgaaagatc ccaacgaaaa gagagaccac atggtccttc ttgagtttgt aacagctgat
     1501 gggattacac atggcatgga tgaactatac aaatgatcta gaactatagt gagtcgtatt
     1561 acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1621 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1681 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     1741 gggaggtgtg ggaggttttt taattgatat ccgcggcgcc aatgcattgg gcccggtacc
     1801 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1861 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     1921 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     1981 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2041 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2101 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2161 tccattttcg gatctgatca gcacgtgatg aaaaagcctg aactcaccgc gacgtctgtc
     2221 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     2281 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     2341 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
     2401 ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc
     2461 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
     2521 ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc
     2581 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     2641 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     2701 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     2761 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     2821 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     2881 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     2941 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt
     3001 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     3061 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     3121 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     3181 cgccccagca ctcgtccgag ggcaaaggaa tagcacgtgc tacgagattt cgattccacc
     3241 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     3301 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     3361 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3421 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     3481 tacgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3541 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3601 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3661 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3721 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3781 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3841 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3901 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3961 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4021 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4081 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4141 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4201 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4261 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4321 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     4381 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4441 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4501 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4561 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4621 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4681 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4741 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4801 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4861 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4921 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4981 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5041 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5101 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5161 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5221 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5281 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5341 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5401 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5461 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5521 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5581 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5641 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     5701 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5761 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5821 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5881 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5941 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     6001 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     6061 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     6121 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     6181 tcgaccatag ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc
     6241 tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc
     6301 tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggact
     6361 tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc
     6421 ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat
     6481 tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc
     6541 tattggccag gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt
     6601 tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct
     6661 tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt
     6721 cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca
     6781 attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat
     6841 tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt
     6901 caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa
     6961 gtacattact attggaagga cgccagggta cattggcagt actcccattg acgtcaatgg
     7021 cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca
     7081 atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc
     7141 tatataagca atgctcgttt agggaac
//
