LOCUS Exported 7717 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3.1 FLIPE-600n SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7717) AUTHORS Okumoto S, Looger LL, Micheva KD, Reimer RJ, Smith SJ, Frommer WB TITLE Detection of glutamate release from neurons by genetically encoded surface-displayed FRET nanosensors. JOURNAL Proc Natl Acad Sci U S A. 2005 Jun 14. 102(24):8740-5. PUBMED 15939876 REFERENCE 2 (bases 1 to 7717) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7717 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 990..1706 /codon_start=1 /product="enhanced CFP" /label=ECFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2565..3284 /codon_start=1 /product="yellow fluorescent protein (YFP) with fast and efficient maturation (Nagai et al., 2002)" /label=Venus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3311..3328) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3317..3541 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3587..4015 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3674..3693) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3884..3905 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4024..4044) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4029..4358 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4209..4344 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4271..4290 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4425..5219 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4479..4498) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5089..5108 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5393..5514 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5430..5449) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5484..5503 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5563..5579) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5563..5579) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5576..5598) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5587..5603 /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(5611..5641) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5656..5677 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5794..5811) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5965..6550) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6045..6064) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6721..7581) /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 7344..7363 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7582..7686) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcc 961 accacactgg actagtggat ccgggccgca tggtgagcaa gggcgaggag ctgttcaccg 1021 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt 1081 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca 1141 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt 1201 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg 1261 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg 1321 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact 1381 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacatc agccacaacg 1441 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca 1501 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 1561 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag 1621 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca 1681 ctctcggcat ggacgagctg tacaagggtg gtaccggagg cgccgcaggc agcacgctgg 1741 acaaaatcgc caaaaacggt gtgattgtcg tcggtcaccg tgaatcttca gtgcctttct 1801 cttattacga caatcagcaa aaagtggtgg gttactcgca ggattactcc aacgccattg 1861 ttgaagcagt gaaaaagaaa ctcaacaaac cggacttgca ggtaaaactg attccgatta 1921 cctcacaaaa ccgtattcca ctgctgcaaa acggcacttt cgattttgaa tgtggttcta 1981 ccaccaacaa cgtcgaacgc caaaaacagg cggctttctc tgacactatt ttcgtggtcg 2041 gtacgcgcct gttgaccaaa aagggtggcg atatcaaaga ttttgccaac ctgaaagaca 2101 aagccgtagt cgtcacttcc ggcactacct ctgaagtttt gctcaacaaa ctgaatgaag 2161 agcaaaaaat gaatatgcgc atcatcagcg ccaaagatca cggtgactct ttccgcaccc 2221 tggaaagcgg tcgtgccgtt gcctttatga tggatgacgc tctgctggcc ggtgaacgtg 2281 cgaaagcgaa gaaaccagac aactgggaaa tcgtcggcaa gccgcagtct caggaggcct 2341 acggttgtat gttgcgtaaa gatgatccgc agttcaaaaa gctgatggat gacaccatcg 2401 ctcaggtgca gacctccggt gaagcggaaa aatggtttga taagtggttc aaaaatccaa 2461 ttccgccgaa aaacctgaac atgaatttcg aactgtcaga cgaaatgaaa gcactgttca 2521 aagaaccgaa tgacaaggca ctgaacggcg ccggtaccgg tggaatggtg agcaagggcg 2581 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc 2641 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga 2701 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg 2761 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca 2821 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca 2881 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc 2941 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact 3001 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact 3061 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga 3121 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt 3181 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga 3241 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaaagctt aagtttaaac 3301 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 3361 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 3421 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 3481 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 3541 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 3601 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 3661 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 3721 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 3781 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 3841 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 3901 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 3961 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 4021 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 4081 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 4141 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 4201 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 4261 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 4321 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 4381 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 4441 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 4501 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 4561 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 4621 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 4681 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 4741 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 4801 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 4861 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 4921 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 4981 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 5041 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 5101 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 5161 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 5221 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 5281 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 5341 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 5401 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 5461 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 5521 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 5581 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 5641 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 5701 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 5761 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 5821 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 5881 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 5941 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 6001 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 6061 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 6121 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 6181 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 6241 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 6301 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 6361 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 6421 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 6481 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 6541 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 6601 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 6661 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 6721 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 6781 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 6841 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 6901 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 6961 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 7021 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 7081 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 7141 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 7201 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 7261 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 7321 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 7381 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 7441 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 7501 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 7561 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 7621 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 7681 tccgcgcaca tttccccgaa aagtgccacc tgacgtc //