LOCUS Exported 7143 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS ArcLight-A242 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7143) 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 7143) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7143 /organism="synthetic DNA construct" /mol_type="other DNA" 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" promoter complement(1523..1540) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" 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 /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(3529..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(4642..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 6166..7143 /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 cgggagcata 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 aagaacagta 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 cgaccgccaa ttcaatatgg 6181 cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg 6241 cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg 6301 cggacttggc accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg 6361 gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat 6421 attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata 6481 taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt 6541 attggcccta tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc 6601 tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc atagccactc 6661 ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc 6721 ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc 6781 tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg 6841 cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg 6901 gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc 6961 agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc 7021 caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg 7081 acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg 7141 aac //