LOCUS V019087 10454 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019087 VERSION V019087 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..5134 /label="GRIN2B(NM_000834)" /note="GRIN2B(NM_000834)" /gene="GRIN2B" CDS 5141..5206 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 5230..5818 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 5884..5932 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 6134..6562 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 6576..6905 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 6972..7766 /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 7942..8063 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(8112..8128) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8136..8152 /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(8160..8190) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8205..8226 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8514..9102) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9273..10133) /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(10134..10238) /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 ccatgaagcc cagagcggag tgctgttctc ccaagttctg 721 gttggtgttg gccgtcctgg ccgtgtcagg cagcagagct cgttctcaga agagcccccc 781 cagcattggc attgctgtca tcctcgtggg cacttccgac gaggtggcca tcaaggatgc 841 ccacgagaaa gatgatttcc accatctctc cgtggtaccc cgggtggaac tggtagccat 901 gaatgagacc gacccaaaga gcatcatcac ccgcatctgt gatctcatgt ctgaccggaa 961 gatccagggg gtggtgtttg ctgatgacac agaccaggaa gccatcgccc agatcctcga 1021 tttcatttca gcacagactc tcacccccat cctgggcatc cacgggggct cctctatgat 1081 aatggcagat aaggatgaat cctccatgtt cttccagttt ggcccatcaa ttgaacagca 1141 agcttccgta atgctcaaca tcatggaaga atatgactgg tacatctttt ctatcgtcac 1201 cacctatttc cctggctacc aggactttgt aaacaagatc cgcagcacca ttgagaatag 1261 ctttgtgggc tgggagctag aggaggtcct cctactggac atgtccctgg acgatggaga 1321 ttctaagatc cagaatcagc tcaagaaact tcaaagcccc atcattcttc tttactgtac 1381 caaggaagaa gccacctaca tctttgaagt ggccaactca gtagggctga ctggctatgg 1441 ctacacgtgg atcgtgccca gtctggtggc aggggataca gacacagtgc ctgcggagtt 1501 ccccactggg ctcatctctg tatcatatga tgaatgggac tatggcctcc ccgccagagt 1561 gagagatgga attgccataa tcaccactgc tgcttctgac atgctgtctg agcacagctt 1621 catccctgag cccaaaagca gttgttacaa cacccacgag aagagaatct accagtccaa 1681 tatgctaaat aggtatctga tcaatgtcac ttttgagggg aggaatttgt ccttcagtga 1741 agatggctac cagatgcacc cgaaactggt gataattctt ctgaacaagg agaggaagtg 1801 ggaaagggtg gggaagtgga aagacaagtc cctgcagatg aagtactatg tgtggccccg 1861 aatgtgtcca gagactgaag agcaggagga tgaccatctg agcattgtga ccctggagga 1921 ggcaccattt gtcattgtgg aaagtgtgga ccctctgagt ggaacctgca tgaggaacac 1981 agtcccctgc caaaaacgca tagtcactga gaataaaaca gacgaggagc cgggttacat 2041 caaaaaatgc tgcaaggggt tctgtattga catccttaag aaaatttcta aatctgtgaa 2101 gttcacctat gacctttacc tggttaccaa tggcaagcat gggaagaaaa tcaatggaac 2161 ctggaatggt atgattggag aggtggtcat gaagagggcc tacatggcag tgggctcact 2221 caccatcaat gaggaacgat cggaggtggt cgacttctct gtgcccttca tagagacagg 2281 catcagtgtc atggtgtcac gcagcaatgg gactgtctca ccttctgcct tcttagagcc 2341 attcagcgct gacgtatggg tgatgatgtt tgtgatgctg ctcatcgtct cagccgtggc 2401 tgtctttgtc tttgagtact tcagccctgt gggttataac aggtgcctcg ctgatggcag 2461 agagcctggt ggaccctctt tcaccatcgg caaagctatt tggttgctct ggggtctggt 2521 gtttaacaac tccgtacctg tgcagaaccc aaaggggacc acctccaaga tcatggtgtc 2581 agtgtgggcc ttctttgctg tcatcttcct ggccagctac actgccaact tagctgcctt 2641 catgatccaa gaggaatatg tggaccaggt ttctggcctg agcgacaaaa agttccagag 2701 acctaatgac ttctcacccc ctttccgctt tgggaccgtg cccaacggca gcacagagag 2761 aaatattcgc aataactatg cagaaatgca tgcctacatg ggaaagttca accagagggg 2821 tgtagatgat gcattgctct ccctgaaaac agggaaactg gatgccttca tctatgatgc 2881 agcagtgctg aactatatgg caggcagaga tgaaggctgc aagctggtga ccattggcag 2941 tgggaaggtc tttgcttcca ctggctatgg cattgccatc caaaaagatt ctgggtggaa 3001 gcgccaggtg gaccttgcta tcctgcagct ctttggagat ggggagatgg aagaactgga 3061 agctctctgg ctcactggca tttgtcacaa tgagaagaat gaggtcatga gcagccagct 3121 ggacattgac aacatggcag gggtcttcta catgttgggg gcggccatgg ctctcagcct 3181 catcaccttc atctgcgaac accttttcta ttggcagttc cgacattgct ttatgggtgt 3241 ctgttctggc aagcctggca tggtcttctc catcagcaga ggtatctaca gctgcatcca 3301 tggggtggcg atcgaggagc gccagtctgt aatgaactcc cccaccgcaa ccatgaacaa 3361 cacacactcc aacatcctgc gcctgctgcg cacggccaag aacatggcta acctgtctgg 3421 tgtgaatggc tcaccgcaga gcgccctgga cttcatccga cgggagtcat ccgtctatga 3481 catctcagag caccgccgca gcttcacgca ttctgactgc aaatcctaca acaacccgcc 3541 ctgtgaggag aacctcttca gtgactacat cagtgaggta gagagaacgt tcgggaacct 3601 gcagctgaag gacagcaacg tgtaccaaga tcactaccac catcaccacc ggccccatag 3661 tattggcagt gccagctcca tcgatgggct ctacgactgt gacaacccac ccttcaccac 3721 ccagtccagg tccatcagca agaagcccct ggacatcggc ctcccctcct ccaagcacag 3781 ccagctcagt gacctgtacg gcaaattctc cttcaagagc gaccgctaca gtggccacga 3841 cgacttgatc cgctccgatg tctctgacat ctcaacccac accgtcacct atgggaacat 3901 cgagggcaat gccgccaaga ggcgtaagca gcaatataag gacagcctga agaagcggcc 3961 tgcctcggcc aagtcccgca gggagtttga cgagatcgag ctggcctacc gtcgccgacc 4021 gccccgctcc cctgaccaca agcgctactt cagggacaag gaagggctac gggacttcta 4081 cctggaccag ttccgaacaa aggagaactc accccactgg gagcacgtag acctgaccga 4141 catctacaag gagcggagtg atgactttaa gcgcgactcc gtcagcggag gagggccctg 4201 taccaacagg tctcacatca agcacgggac gggcgacaaa cacggcgtgg tcagcggggt 4261 acctgcacct tgggagaaga acctgaccaa cgtggagtgg gaggaccggt ccgggggcaa 4321 cttctgccgc agctgtccct ccaagctgca caactactcc acgacggtga cgggtcagaa 4381 ctcgggcagg caggcgtgca tccggtgtga ggcttgcaag aaagcaggca acctgtatga 4441 catcagtgag gacaactccc tgcaggaact ggaccagccg gctgccccag tggcggtgac 4501 gtcaaacgcc tccaccacta agtaccctca gagcccgact aattccaagg cccagaagaa 4561 gaaccggaac aaactgcgcc ggcagcactc ctacgacacc ttcgtggacc tgcagaagga 4621 agaagccgcc ctggccccgc gcagcgtaag cctgaaagac aagggccgat tcatggatgg 4681 gagcccctac gcccacatgt ttgagatgtc agctggcgag agcacctttg ccaacaacaa 4741 gtcctcagtg cccactgccg gacatcacca ccacaacaac cccggcggcg ggtacatgct 4801 cagcaagtcg ctctaccctg accgggtcac gcaaaaccct ttcatcccca cttttgggga 4861 cgaccagtgc ttgctccatg gcagcaaatc ctacttcttc aggcagccca cggtggcggg 4921 ggcgtcgaaa gccaggccgg acttccgggc ccttgtcacc aacaagccgg tggtctcggc 4981 ccttcatggg gccgtgccag cccgtttcca gaaggacatc tgtataggga accagtccaa 5041 cccctgtgtg cctaacaaca aaaaccccag ggctttcaat ggctccagca atgggcatgt 5101 ttatgagaaa ctttctagta ttgagtctga tgtcctcgag gactacaaag accatgacgg 5161 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga 5221 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 5281 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 5341 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 5401 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 5461 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 5521 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 5581 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca 5641 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 5701 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 5761 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 5821 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta 5881 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc 5941 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa 6001 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg 6061 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct 6121 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 6181 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 6241 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta 6301 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 6361 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 6421 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 6481 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt 6541 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 6601 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 6661 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 6721 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 6781 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 6841 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 6901 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 6961 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 7021 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 7081 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 7141 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 7201 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 7261 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 7321 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 7381 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 7441 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 7501 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 7561 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 7621 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 7681 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 7741 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa 7801 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 7861 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 7921 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 7981 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 8041 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 8101 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 8161 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 8221 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 8281 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 8341 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 8401 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 8461 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 8521 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 8581 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 8641 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 8701 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 8761 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 8821 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 8881 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 8941 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 9001 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 9061 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 9121 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 9181 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 9241 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 9301 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 9361 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 9421 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 9481 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 9541 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 9601 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 9661 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 9721 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 9781 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 9841 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 9901 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 9961 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 10021 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 10081 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 10141 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 10201 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 10261 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct 10321 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 10381 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 10441 agaatctgct tagg //