LOCUS V019146 10394 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019146 VERSION V019146 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 683..5074 /label="Grin2a(NM_012573)" /note="Grin2a(NM_012573)" /gene="Grin2a" CDS 5081..5146 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 5170..5758 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 5824..5872 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 6074..6502 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 6516..6845 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 6912..7706 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 7882..8003 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(8052..8068) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8076..8092 /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(8100..8130) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8145..8166 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8454..9042) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9213..10073) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(10074..10178) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccgcca ccatgggcag attgggctac tggaccttgc tggtattgcc 721 ggcccttctg gtctggcgcg atccggcgca gaacgcggcg gcggagaagg gtcctccagc 781 gctgaacatt gcggtgctgc tgggtcacag ccacgacgtg acagaacgcg aacttcgaaa 841 tctgtggggc ccagagcagg caaccggctt gcccctggat gtgaacgtgg tggcgttatt 901 gatgaaccgc actgacccta agagcctcat cacgcatgtg tgcgacctca tgtccggggc 961 gcgcatccac ggcttggtgt ttggagatga cacggaccag gaggctgtgg cccagatgct 1021 ggattttatc tcctcacaga cttttatccc catcttgggc attcatgggg gtgcatctat 1081 gatcatggct gacaaggatc cgacatccac gttcttccag tttggagcct ccatccagca 1141 gcaagccaca gttatgctga agatcatgca ggactacgac tggcacgtct tctccctggt 1201 caccaccatc ttccctggct accgagactt catcagcttt atcaagacaa cagtggacaa 1261 cagctttgtg ggctgggata tgcagaacgt gatcacactg gacacctcct tcgaggacgc 1321 caagacgcag gtccagctga agaagatcca ttcttctgtc atcctgctct actgctccaa 1381 agacgaggct gtcctcatcc tgagcgaggc tcgctccctc ggcctcactg gctatgattt 1441 cttctggatt gtccccagtt tggtgtctgg gaacacagag ctcatcccca aagagtttcc 1501 atcaggtctc atttcagtct cttatgacga ctgggactac agcctggagg caagagtgag 1561 agacggtctt gggatcttaa ccactgccgc atcctccatg ttggagaagt tctcctacat 1621 tcctgaggcc aaggccagct gctatgggca ggcagagaag ccagagaccc cgctacacac 1681 cctgcaccaa ttcatggtca atgtgacttg ggatggcaag gacttgtcct tcactgagga 1741 aggttaccag gtgcacccca ggcttgtggt gatcgtgctg aacaaggacc gggagtggga 1801 aaaggtgggc aagtgggaga accagacgct gagcctgagg cacgctgtgt ggccaaggta 1861 caagtccttt tctgactgcg agccagatga caaccacctc agcattgtca ccttggagga 1921 agcccccttc gtcatcgtag aggacataga ccccctgact gagacctgtg tgaggaacac 1981 ggtgccctgt cggaagttcg tcaagatcaa caattcaacc aacgaaggga tgaatgtgaa 2041 gaaatgctgc aaggggttct gcatcgacat cctcaagaag ctgtccagaa ctgtgaagtt 2101 cacctatgac ctctacctgg tgaccaatgg gaagcatggg aaaaaggtta acaatgtgtg 2161 gaatggaatg ataggtgaag tggtctatca acgagcagtc atggctgtgg gctcactcac 2221 catcaatgag gagcgttcgg aagtggtgga cttctcggtg cccttcgtgg agacaggaat 2281 cagcgtcatg gtctccagga gtaatggcac tgtctcccct tctgctttcc tcgaaccctt 2341 cagtgcctcc gtctgggtga tgatgtttgt gatgctgctc atcgtctcag ccattgctgt 2401 cttcgttttt gaatacttca gtcctgttgg atacaacaga aacttagcca aagggaaagc 2461 tccccacggg ccttctttta ctattggaaa agctatatgg ctcctctggg gcctggtctt 2521 caacaattct gtgcctgtcc agaatcctaa aggcacaacc agcaagatca tggtgtcagt 2581 gtgggccttc tttgctgtca tcttcctggc cagttacaca gccaacctgg ctgccttcat 2641 gatccaggag gagtttgtgg accaagtgac tggcctcagt gacaagaagt tccagagacc 2701 tcatgactat tctccacctt tccgatttgg gacggtaccc aatggaagta cagagaggaa 2761 tattcgtaac aactacccgt atatgcacca gtacatgacc agattcaacc agaggggagt 2821 ggaggatgcc ttggtcagct tgaaaaccgg gaagttggac gctttcatct atgacgcagc 2881 cgtcttgaac tacaaggccg ggagggatga aggctgtaaa ctggtgacca ttgggagcgg 2941 gtacatcttt gctaccacag gctatggaat tgcgctgcag aagggctcac cctggaagag 3001 gcagattgac ctcgctctgc tccagtttgt tggtgatggt gagatggagg agctggagac 3061 actgtggctt acgggcatct gccacaacga gaagaatgag gtgatgagta gccagctgga 3121 catcgataac atggcgggcg tgttctacat gctggctgca gccatggccc tcagcctcat 3181 caccttcatc tgggagcacc tcttctactg gaagctgcgc ttctgcttca caggcgtgtg 3241 ctctgaccgg cccgggctgc tcttctccat cagcaggggc atctatagtt gcatccatgg 3301 ggtacacatt gaagaaaaga agaaatctcc agatttcaat ctgactggat cacagagcaa 3361 catgctaaag cttcttcggt cagctaaaaa catctccaat atgtccaaca tgaactcctc 3421 aagaatggac tcacctaaaa gagctactga cttcattcaa agagggtcac ttattgtgga 3481 catggtttca gacaagggaa atttgatata ttcagacaac agatcctttc aagggaagga 3541 cagtatattt ggagacaaca tgaatgaact ccaaacattt gtggccaaca ggcacaagga 3601 taatctcagt aactatgtgt ttcaaggaca gcatcctctc actctcaatg agtccaaccc 3661 taacacagta gaggtggctg tcagcactga atccaaaggg aactcccgac cccggcagct 3721 ttggaagaaa tccatggagt ctctacgcca ggattctcta aaccagaacc cagtctccca 3781 gagggatgag aagactgcag agaatcggac ccactcgcta aagagtccta ggtatcttcc 3841 agaagaggta gcccactctg acatttcaga aacctcaagc cgggccacat gccacaggga 3901 gccagataac aataagaacc acaagaccaa ggataacttc aaacggtcaa tggcctctaa 3961 gtatcccaag gactgtagcg atgttgaccg cacctacatg aaaaccaaag caagttctcc 4021 cagggataag atctatacca ttgatggtga gaaggagccc agcttccact tagatcctcc 4081 tcagtttgtt gagaatataa ccctgcctga gaatgtgggc ttcccagata cctaccaaga 4141 tcacaatgag aacttccgca agggggactc cacactgccc atgaacagga acccattaca 4201 taatgaagac gggcttccca acaatgacca atataaactc tatgccaagc actttacctt 4261 gaaagacaag ggttccccac acagtgaggg cagtgatcga taccggcaga actccacaca 4321 ttgcagaagc tgcctttcga atctgcccac ctactcaggc cactttacca tgaggtctcc 4381 tttcaagtgt gatgcctgtc tgcggatggg gaatctctat gacattgatg aagaccagat 4441 gcttcaggag acaggtaacc cagctactcg ggaggaggtc taccagcagg actggtcaca 4501 gaacaacgcc ctccagttcc agaagaacaa gctaaggatt aaccgacagc actcctatga 4561 taacattctg gacaaaccca gagagataga ccttagcagg ccctcccgga gcataagcct 4621 caaggacagg gaacggctac tggagggcaa cttgtatggg agcctgttca gtgtcccctc 4681 aagcaaactc ttggggaaca aaagctccct tttcccccaa ggtctggagg acagcaagag 4741 gagcaagtct ctcttgccag accacgcctc cgataatcct ttcctccaca cgtatgggga 4801 tgaccaacgc ttagttatcg ggagatgtcc ctcggaccct tacaaacact cattgccatc 4861 acaggcggta aatgacagct atcttcggtc atccttgagg tcaacagcat catattgctc 4921 cagggacagt cggggccaca gtgatgtgta tatttcagag catgttatgc cttatgctgc 4981 aaataagaat accatgtact ctacccccag ggttttaaat tcctgcagca atagacgagt 5041 gtacaagaaa atgcctagta tcgaatctga tgttctcgag gactacaaag accatgacgg 5101 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga 5161 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 5221 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 5281 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 5341 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 5401 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 5461 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 5521 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca 5581 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 5641 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 5701 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 5761 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta 5821 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc 5881 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa 5941 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg 6001 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct 6061 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 6121 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 6181 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta 6241 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 6301 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 6361 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 6421 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt 6481 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 6541 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 6601 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 6661 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 6721 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 6781 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 6841 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 6901 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 6961 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 7021 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 7081 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 7141 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 7201 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 7261 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 7321 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 7381 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 7441 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 7501 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 7561 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 7621 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 7681 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa 7741 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 7801 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 7861 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 7921 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 7981 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 8041 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 8101 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 8161 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 8221 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 8281 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 8341 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 8401 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 8461 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 8521 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 8581 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 8641 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 8701 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 8761 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 8821 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 8881 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 8941 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 9001 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 9061 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 9121 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 9181 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 9241 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 9301 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 9361 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 9421 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 9481 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 9541 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 9601 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 9661 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 9721 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 9781 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 9841 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 9901 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 9961 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 10021 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 10081 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 10141 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 10201 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct 10261 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 10321 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 10381 agaatctgct tagg //