LOCUS V032787 7033 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032787 VERSION V032787 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 964..2523 /label="SARS2(NM_001145901)" /note="SARS2(NM_001145901)" /gene="SARS2" CDS 2530..2595 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2633..2857 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2903..3331 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3345..3674 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3741..4535 /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 4709..4830 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4879..4895) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4903..4919 /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(4927..4957) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4972..4993 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5281..5866) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6037..6897) /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(6898..7002) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatggctg cgtccatggc gcggcgcttg tggcctttgc tgactcgtcg ggggttccgg 1021 ccccggggag gctgcatctc caacgatagt ccaaggagaa gtttcactac agagaaacga 1081 aaccggaacc tcctgtacga gtatgcgcgc gagggctaca gcgcactccc tcagctggac 1141 atagagcggt tctgcgcatg cccagaagag gccgcacacg ccctggagct ccgcaagggg 1201 gagctgcgct cggcggacct gcccgcgatc atctcgacat ggcaggagct gaggcagctg 1261 caggagcaga tccggagcct ggaggaagag aaggcagctg tgactgaggc agtgcgggcc 1321 ctgctggcaa accaggacag tggtgaagtg cagcaggtgc ggctggatcc aggtgctggc 1381 tccatatttg gtcctacgtt cctcccattc ccaggccagc tttctctcct tgtggaggcc 1441 cagcttgagg agcagttcta cctgcaggcg ctgaagctgc ccaaccagac ccacccagac 1501 gtgcccgtcg gggatgagag ccaggctcga gtgctccaca tggtcggaga caagccagtt 1561 ttctccttcc aacctcgggg ccacctggaa attggcgaga aactcgacat catccgtcag 1621 aagcgcctgt cccacgtgtc tggccaccgg tcctattacc tgcgcggggc tggagccctc 1681 ctgcagcacg gcctggtcaa cttcacattc aacaagcttc tccgccgggg cttcaccccc 1741 atgacggtgc cagaccttct ccgcggagca gtgtttgaag gctgtgggat gacaccaaat 1801 gccaacccat cccaaattta caacatcgac cctgcccgct tcaaagatct caacctggct 1861 ggaacagcgg aggtggggct tgcaggctac ttcatggacc acaccgtggc cttcagggac 1921 ctgccagtca ggatggtttg ctccagcacc tgctaccggg cagagacaaa cacgggacag 1981 gaaccccggg ggctgtatcg agtacaccac ttcaccaagg tggagatgtt tggggtgaca 2041 ggccctgggc tggagcagag ctcacagctg ctggaggagt tcctgtccct tcagatggag 2101 atcttgacag agctgggctt gcacttccgg gtcctggata tgcccaccca agaactgggc 2161 ctccccgcct accgcaagtt tgacattgag gcctggatgc caggccgagg ccgctttgga 2221 gaggtcacca gtgcttccaa ctgcacagac ttccagagcc gccgcctcca catcatgttc 2281 cagaccgagg ctggggagct gcagtttgcc cacacggtga acgccaccgc ctgtgctgtc 2341 ccccgccttc tcatcgcgct cctggagagt aaccagcaga aggacggctc agtgctcgtg 2401 ccccctgccc tccagtccta cctcggcact gatcggatca cagcccctac ccacgtgcct 2461 ctccagtaca tcggccccaa ccagccccgg aagcctgggc tgcctggcca gcctgctgta 2521 agcctcgagg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag 2581 gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc gactgtgcct 2641 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt 2701 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg 2761 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac 2821 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc 2881 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 2941 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 3001 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 3061 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 3121 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 3181 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 3241 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat 3301 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt 3361 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3421 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3481 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 3541 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 3601 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 3661 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga 3721 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg 3781 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 3841 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt 3901 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg 3961 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 4021 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 4081 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 4141 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 4201 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc 4261 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc 4321 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 4381 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 4441 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 4501 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac 4561 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga 4621 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga 4681 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa 4741 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 4801 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta 4861 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 4921 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 4981 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 5041 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 5101 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 5161 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 5221 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 5281 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 5341 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 5401 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 5461 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 5521 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 5581 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 5641 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 5701 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 5761 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt 5821 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 5881 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 5941 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 6001 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 6061 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 6121 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 6181 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 6241 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 6301 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 6361 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 6421 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 6481 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 6541 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 6601 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 6661 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 6721 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 6781 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 6841 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 6901 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 6961 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 7021 gccacctgac gtc //