LOCUS V021340 7886 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021340 VERSION V021340 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..2533 /label="Scg2(NM_009129)" /note="Scg2(NM_009129)" /gene="Scg2" CDS 2573..2638 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2662..3250 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3316..3364 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3566..3994 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4008..4337 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4404..5198 /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 5374..5495 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5544..5560) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5568..5584 /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(5592..5622) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5637..5658 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5946..6534) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6705..7565) /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(7566..7670) /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 ccatggctgg agctaaggcg taccgacttg gagcagttct 721 gcttcttatc cacttaattt tcctcatctc tggagccgaa gcagcttcct tccagcgaaa 781 ccagctgctt cagaaagaac cagacctcag attggagaat gtccaaaagt ttcctagtcc 841 agaaatgatc agggctttgg agtacataga aaagctcagg cagcaagctc acagagaaga 901 aagcagccca gactacaatc cctaccaagg cgtctctgtt cctcttcaac tcaaagaaaa 961 cggagaagaa agccacttgg cagagagctc aagggatgca ctgagtgaag acgagtggat 1021 gcggataata ctcgaggctc tgaggcaggc tgaaaatgag ccgccatctg cccccaaaga 1081 gaacaagccc tatgccttga atctggagaa gaacttccca gtggacacgc ctgatgacta 1141 tgagactcaa cagtggcctg agaggaaact caagcacatg cggttccctc tcatgtatga 1201 agagaattcc agagaaaacc ccttcaaacg cacaaatgaa atagtcgagg aacaatacac 1261 accccaaagt cttgctaccc tggagtctgt gttccaagag cttgggaaac tgacagggcc 1321 aagcaaccag aagcgtgaga gggttgacga ggaacaaaag ctgtacacag atgatgaaga 1381 cgacgtgtac aagaccaaca acattgccta tgaagatgtc gtggggggag aagactggag 1441 ccccatagag gagaaaatag agactcaaac ccaggaagag gtgagagaca gcaaagagaa 1501 cacagaaaaa aatgaacaaa tcaatgaaga gatgaaacgt tcagggcagt tggggctccc 1561 agatgaagaa aaccggagag agagtaaaga ccaactctca gaggatgcct ccaaagttat 1621 cacctacctg agaaggttag tgaatgctgt gggcagtggg aggtcacaga gtgggccaaa 1681 tggggacagg gcagcccggc ttcttcagaa gccccttgat tctcagtcta tttatcagct 1741 gattgaaatc tccaggaatt tgcagatacc ccctgaagat ttaattgaga tgctcaaagc 1801 tggagagaag ccaaatgggt tggtggagcc agagcaggat ctggagcttg ctgttgacct 1861 agatgacatc ccagaggctg acctagaccg tccagacatg tttcaaagta agatgctctc 1921 caagggtggg tatcccaagg cacctggtcg tggtatggta gaggccttgc ctgatgggct 1981 gagtgtcgag gacattttaa atgttttagg gatggagaat gtagtaaatc agaagtcccc 2041 atattttccc aaccaatata gccaagacaa ggctctgatg aggctccctt atggtcctgg 2101 gaaatctaga gccaaccaga ttcccaaagt agcctggatc cctgatgttg aaagcagaca 2161 agcaccttat gaaaatctga atgaccaaga attgggagag tacttagcca ggatgctagt 2221 taagtaccct gagctcctga ataccaacca gctgaagaga gtgcccagtc cagtctcctc 2281 agaggatgac ctccaagaag aagagcagct cgagcaggcc atcaaggaac atctggggcc 2341 aggaagctcc caggaaatgg agagactggc caaggtgagc aaaaggatcc ccgtaggatc 2401 cctgaagaat gaggacaccc caaacagaca gtacctggat gaagatatgc tcctgaaagt 2461 gctggagtac ctcaaccaag agcaggcaga gcaggggagg gagcatcttg ccaagcgggc 2521 catggaaaac atggaattct gcagatatcc agcacagtgg cggccgctcg aggactacaa 2581 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 2641 gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact 2701 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 2761 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 2821 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 2881 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 2941 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 3001 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg 3061 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 3121 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 3181 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 3241 gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag 3301 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt 3361 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc 3421 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt 3481 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc 3541 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 3601 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 3661 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 3721 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 3781 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 3841 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 3901 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa 3961 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 4021 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 4081 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 4141 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 4201 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 4261 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 4321 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 4381 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 4441 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 4501 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 4561 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 4621 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 4681 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 4741 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 4801 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 4861 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 4921 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 4981 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 5041 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 5101 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 5161 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa 5221 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc 5281 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc 5341 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt 5401 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 5461 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 5521 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 5581 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 5641 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 5701 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 5761 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 5821 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 5881 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 5941 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 6001 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 6061 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 6121 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 6181 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 6241 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 6301 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 6361 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 6421 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 6481 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 6541 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 6601 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 6661 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 6721 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 6781 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 6841 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 6901 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 6961 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 7021 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 7081 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 7141 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 7201 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 7261 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 7321 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 7381 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 7441 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 7501 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 7561 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 7621 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 7681 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc 7741 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt 7801 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg 7861 acaattgcat gaagaatctg cttagg //