LOCUS V029285 6985 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029285 VERSION V029285 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..2517 /label="SARS2(NM_017827)" /note="SARS2(NM_017827)" /gene="SARS2" CDS 2524..2547 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" polyA_signal 2585..2809 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2855..3283 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3297..3626 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3693..4487 /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 4661..4782 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4831..4847) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4855..4871 /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(4879..4909) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4924..4945 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5233..5818) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5989..6849) /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(6850..6954) /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 cagcaggacc ccaagtacca gggtctgcgg 1381 gcacgtggcc gggagatccg gaaggagctt gttcacctgt accccaggga ggcccagctt 1441 gaggagcagt tctacctgca ggcgctgaag ctgcccaacc agacccaccc agacgtgccc 1501 gtcggggatg agagccaggc tcgagtgctc cacatggtcg gagacaagcc agttttctcc 1561 ttccaacctc ggggccacct ggaaattggc gagaaactcg acatcatccg tcagaagcgc 1621 ctgtcccacg tgtctggcca ccggtcctat tacctgcgcg gggctggagc cctcctgcag 1681 cacggcctgg tcaacttcac attcaacaag cttctccgcc ggggcttcac ccccatgacg 1741 gtgccagacc ttctccgcgg agcagtgttt gaaggctgtg ggatgacacc aaatgccaac 1801 ccatcccaaa tttacaacat cgaccctgcc cgcttcaaag atctcaacct ggctggaaca 1861 gcggaggtgg ggcttgcagg ctacttcatg gaccacaccg tggccttcag ggacctgcca 1921 gtcaggatgg tttgctccag cacctgctac cgggcagaga caaacacggg acaggaaccc 1981 cgggggctgt atcgagtaca ccacttcacc aaggtggaga tgtttggggt gacaggccct 2041 gggctggagc agagctcaca gctgctggag gagttcctgt cccttcagat ggagatcttg 2101 acagagctgg gcttgcactt ccgggtcctg gatatgccca cccaagaact gggcctcccc 2161 gcctaccgca agtttgacat tgaggcctgg atgccaggcc gaggccgctt tggagaggtc 2221 accagtgctt ccaactgcac agacttccag agccgccgcc tccacatcat gttccagacc 2281 gaggctgggg agctgcagtt tgcccacacg gtgaacgcca ccgcctgtgc tgtcccccgc 2341 cttctcatcg cgctcctgga gagtaaccag cagaaggacg gctcagtgct cgtgccccct 2401 gccctccagt cctacctcgg cactgatcgg atcacagccc ctacccacgt gcctctccag 2461 tacatcggcc ccaaccagcc ccggaagcct gggctgcctg gccagcctgc tgtaagcctc 2521 gaggattaca aggatgacga cgataagtag tgagggcccg tttaaacccg ctgatcagcc 2581 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 2641 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 2701 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 2761 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 2821 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 2881 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 2941 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 3001 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 3061 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 3121 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 3181 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 3241 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 3301 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3361 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 3421 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 3481 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 3541 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 3601 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat 3661 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 3721 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 3781 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 3841 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc 3901 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 3961 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 4021 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 4081 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 4141 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 4201 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg 4261 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 4321 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 4381 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 4441 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 4501 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc 4561 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 4621 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 4681 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 4741 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 4801 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 4861 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 4921 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 4981 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 5041 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 5101 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 5161 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 5221 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 5281 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 5341 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 5401 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 5461 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 5521 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 5581 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 5641 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 5701 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 5761 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 5821 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 5881 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 5941 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 6001 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 6061 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 6121 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 6181 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 6241 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 6301 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 6361 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 6421 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 6481 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 6541 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 6601 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 6661 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 6721 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 6781 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 6841 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 6901 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 6961 tccccgaaaa gtgccacctg acgtc //