LOCUS V024781 8066 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024781 VERSION V024781 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 941..3556 /label="GRM2(NM_000839)" /note="GRM2(NM_000839)" /gene="GRM2" CDS 3563..3628 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3666..3890 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3936..4364 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4378..4707 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4774..5568 /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 5742..5863 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5912..5928) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5936..5952 /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(5960..5990) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6005..6026 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6314..6899) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7070..7930) /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(7931..8035) /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 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgggatcgc tgcttgcgct 961 cctggcactg ctgctgctgt ggggtgctgt ggctgagggc ccagccaaga aggtgctgac 1021 cctggaggga gacttggtgc tgggtgggct gttcccagtg caccagaagg gcggcccagc 1081 agaggactgt ggtcctgtca atgagcaccg tggcatccag cgcctggagg ccatgctttt 1141 tgcactggac cgcatcaacc gtgacccgca cctgctgcct ggcgtgcgcc tgggtgcaca 1201 catcctcgac agttgctcca aggacacaca tgcgctggag caggcactgg actttgtgcg 1261 tgcctcactc agccgtggtg ctgatggctc acgccacatc tgccccgacg gctcttatgc 1321 gacccatggt gatgctccca ctgccatcac tggtgttatt ggcggttcct acagtgatgt 1381 ctccatccag gtggccaacc tcttgaggct atttcagatc ccacagatta gctacgcctc 1441 taccagtgcc aagctgagtg acaagtcccg ctatgactac tttgcccgca cagtgcctcc 1501 tgacttcttc caagccaagg ccatggctga gattctccgc ttcttcaact ggacctatgt 1561 gtccactgtg gcgtctgagg gcgactatgg cgagacaggc attgaggcct ttgagctaga 1621 ggctcgtgcc cgcaacatct gtgtggccac ctcggagaaa gtgggccgtg ccatgagccg 1681 cgcggccttt gagggtgtgg tgcgagccct gctgcagaag cccagtgccc gcgtggctgt 1741 cctgttcacc cgttctgagg atgcccggga gctgcttgct gccagccagc gcctcaatgc 1801 cagcttcacc tgggtggcca gtgatggttg gggggccctg gagagtgtgg tggcaggcag 1861 tgagggggct gctgagggtg ctatcaccat cgagctggcc tcctacccca tcagtgactt 1921 tgcctcctac ttccagagcc tggacccttg gaacaacagc cggaacccct ggttccgtga 1981 attctgggag cagaggttcc gctgcagctt ccggcagcga gactgcgcag cccactctct 2041 ccgggctgtg ccctttgagc aggagtccaa gatcatgttt gtggtcaatg cagtgtacgc 2101 catggcccat gcgctccaca acatgcaccg tgccctctgc cccaacacca cccggctctg 2161 tgacgcgatg cggccagtta acgggcgccg cctctacaag gactttgtgc tcaacgtcaa 2221 gtttgatgcc ccctttcgcc cagctgacac ccacaatgag gtccgctttg accgctttgg 2281 tgatggtatt ggccgctaca acatcttcac ctatctgcgt gcaggcagtg ggcgctatcg 2341 ctaccagaag gtgggctact gggcagaagg cttgactctg gacaccagcc tcatcccatg 2401 ggcctcaccc tcagccggcc ccctgcccgc ctctcgctgc agtgagccct gcctccagaa 2461 tgaggtgaag agtgtgcagc cgggcgaagt ctgctgctgg ctctgcattc cgtgccagcc 2521 ctatgagtac cgattggacg aattcacttg cgctgattgt ggcctgggct actggcccaa 2581 tgccagcctg actggctgct tcgaactgcc ccaggagtac atccgctggg gcgatgcctg 2641 ggctgtggga cctgtcacca tcgcctgcct cggtgccctg gccaccctct ttgtgctggg 2701 tgtctttgtg cggcacaatg ccacaccagt ggtcaaggcc tcaggtcggg agctctgcta 2761 catcctgctg ggtggtgtct tcctctgcta ctgcatgacc ttcatcttca ttgccaagcc 2821 atccacggca gtgtgtacct tacggcgtct tggtttgggc actgccttct ctgtctgcta 2881 ctcagccctg ctcaccaaga ccaaccgcat tgcacgcatc ttcggtgggg cccgggaggg 2941 tgcccagcgg ccacgcttca tcagtcctgc ctcacaggtg gccatctgcc tggcacttat 3001 ctcgggccag ctgctcatcg tggtcgcctg gctggtggtg gaggcaccgg gcacaggcaa 3061 ggagacagcc cccgaacggc gggaggtggt gacactgcgc tgcaaccacc gcgatgcaag 3121 tatgttgggc tcgctggcct acaatgtgct cctcatcgcg ctctgcacgc tttatgcctt 3181 caagactcgc aagtgccccg aaaacttcaa cgaggccaag ttcattggct tcaccatgta 3241 caccacctgc atcatctggc tggcattcct gcccatcttc tatgtcacct ccagtgacta 3301 ccgggtacag accaccacca tgtgcgtgtc agtcagcctc agcggctccg tggtgcttgg 3361 ctgcctcttt gcgcccaagc tgcacatcat cctcttccag ccgcagaaga acgtggttag 3421 ccaccgggca cccaccagcc gctttggcag tgctgctgcc agggccagct ccagccttgg 3481 ccaagggtct ggctcccagt ttgtccccac tgtttgcaat ggccgtgagg tggtggactc 3541 gacaacgtca tcgcttctcg aggactacaa agaccatgac ggtgattata aagatcatga 3601 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc 3661 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 3721 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 3781 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 3841 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 3901 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 3961 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 4021 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 4081 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 4141 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 4201 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 4261 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 4321 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 4381 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 4441 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 4501 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 4561 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 4621 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 4681 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 4741 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 4801 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 4861 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 4921 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 4981 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 5041 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 5101 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 5161 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 5221 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 5281 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 5341 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 5401 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 5461 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 5521 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 5581 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 5641 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 5701 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 5761 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 5821 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 5881 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 5941 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 6001 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 6061 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 6121 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 6181 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 6241 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 6301 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 6361 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 6421 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 6481 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 6541 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 6601 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 6661 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 6721 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 6781 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 6841 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 6901 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 6961 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 7021 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 7081 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 7141 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 7201 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 7261 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 7321 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 7381 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 7441 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 7501 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 7561 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 7621 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 7681 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 7741 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 7801 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 7861 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 7921 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 7981 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 8041 ttccccgaaa agtgccacct gacgtc //