LOCUS       sequence-46724-e        7146 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 7146)
  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 7146)
  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-A242
FEATURES             Location/Qualifiers
     source          1..7146
                     /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             799..1515
                     /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(829..857)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1452..1472
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    1545..1679
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1784..1804)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1789..2118
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1969..2104
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2031..2050
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2167..3192
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2167..3192
                     /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    3322..3443
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3481..3497)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3481..3497)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3494..3516)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3505..3521
                     /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(3529..3535,3536..3553,3554..3559))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3574..3595
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3712..3729)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3883..4471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3963..3982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4642..5433,5434..5502))
                     /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     5265..5284
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5503..5607)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5633..6088)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5770..5791)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5982..6001
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6162..7146
                     /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 atgaaggctg gggatcccat gagtaaagga gaagaacttt tcactggagt tgtcccaatt
      841 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa
      901 ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct
      961 gttccatggc caacacttgt cactacttta acttatggtg ttcaatgctt ttcaagatac
     1021 ccagatcata tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
     1081 gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt
     1141 gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga
     1201 aacattcttg gacacaaatt ggaatacaac tataacgatc accaggtgta catcatggca
     1261 gacaaacaaa agaatggaat caaagctaac ttcaaaatta gacacaacat tgaagatgga
     1321 ggcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg gcccgtcctt
     1381 ttaccagaca accattacct gtttacaact tctactcttt cgaaagatcc caacgaaaag
     1441 agagaccaca tggtccttct tgagtttgta acagctgatg ggattacaca tggcatggat
     1501 gaactataca aatgatctag aactatagtg agtcgtatta cgtagatcca gacatgataa
     1561 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1621 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     1681 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     1741 aattgatatc cgcggcgcca atgcattggg cccggtaccc tgtggaatgt gtgtcagtta
     1801 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     1861 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     1921 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     1981 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2041 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2101 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
     2161 cacgtgatga aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     2221 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     2281 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     2341 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     2401 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     2461 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     2521 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     2581 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     2641 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     2701 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     2761 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     2821 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     2881 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     2941 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     3001 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     3061 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     3121 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     3181 gcaaaggaat agcacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3241 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3301 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     3361 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3421 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt acgcgcttgg cgtaatcatg
     3481 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3541 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3601 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3721 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3781 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4321 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4561 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4621 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4681 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4741 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4801 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4861 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4921 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4981 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5041 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5101 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5161 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5221 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5281 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5341 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5401 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5461 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5521 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5581 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     5641 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5701 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5761 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5821 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     5881 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     5941 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     6001 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     6061 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
     6121 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgaccatagc caattcaata
     6181 tggcgtatat ggactcatgc caattcaata tggtggatct ggacctgtgc caattcaata
     6241 tggcgtatat ggactcgtgc caattcaata tggtggatct ggaccccagc caattcaata
     6301 tggcggactt ggcaccatgc caattcaata tggcggactt ggcactgtgc caactgggga
     6361 ggggtctact tggcacggtg ccaagtttga ggaggggtct tggccctgtg ccaagtccgc
     6421 catattgaat tggcatggtg ccaataatgg cggccatatt ggctatatgc caggatcaat
     6481 atataggcaa tatccaatat ggccctatgc caatatggct attggccagg ttcaatacta
     6541 tgtattggcc ctatgccata tagtattcca tatatgggtt ttcctattga cgtagatagc
     6601 ccctcccaat gggcggtccc atataccata tatggggctt cctaataccg cccatagcca
     6661 ctcccccatt gacgtcaatg gtctctatat atggtctttc ctattgacgt catatgggcg
     6721 gtcctattga cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct
     6781 ggctcaatgc ccattgacgt caataggacc acccaccatt gacgtcaatg ggatggctca
     6841 ttgcccattc atatccgttc tcacgccccc tattgacgtc aatgacggta aatggcccac
     6901 ttggcagtac atcaatatct attaatagta acttggcaag tacattacta ttggaaggac
     6961 gccagggtac attggcagta ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc
     7021 tgccaagtac atccccattg acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca
     7081 ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct atataagcaa tgctcgttta
     7141 gggaac
//
