LOCUS V020179 7849 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020179 VERSION V020179 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 913..3600 /label="N4BP1(NM_153029)" /note="N4BP1(NM_153029)" /gene="N4BP1" CDS 3613..3678 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3713..3937 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3983..4411 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4425..4754 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4766..5365 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" polyA_signal 5525..5646 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5695..5711) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5719..5735 /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(5743..5773) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5788..5809 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6097..6682) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6853..7713) /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(7714..7818) /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 aagcttgcca ccatggcggc ccgggcggtg ctggacgagt tcactgcgcc agctgagaag 961 gcggagctgc tggagcagag ccgcggccgt atcgagggcc tgtttggcgt gagcctagcc 1021 gtgctcggcg cgctaggggc tgaggagccg ctgcccgcgc gcatctggct gcagctctgc 1081 ggggcgcagg aggcggtgca cagcgccaag gaatatatta aaggaatctg tgaacctgaa 1141 ctagaagaaa gagaatgtta ccccaaggac atgcactgca tttttgttgg ggcagagagc 1201 ctgtttctga aaagcttgat tcaggatact tgtgctgacc tctgcattct ggacattggc 1261 cttcttggca tcagaggaag tgctgaggct gtggtcatgg ctaggagtca cattcaacaa 1321 tttgtaaagc tctttgaaaa taaagagaac ctacccagta gtcagaaaga atcagaggtg 1381 aaaagggaat tcaaacaatt tgttgaagcc catgcagaca attacacaat ggatttgttg 1441 attttgccca cttccttgaa aaaagaactt ttgacactca cacaaggtga ggagaatctc 1501 tttgaaacag gagatgatga ggttattgaa atgagagatt ctcaacaaac agagtttaca 1561 cagaatgctg ccacagggct gaatatttct agagatgaaa ctgttttgca ggaagaggca 1621 agaaataaag ctgggactcc tgtttctgag cttacaaaac aaatggacac agtcctttct 1681 agctcaccag atgtgctttt tgatccaata aatggtctaa ccccagatga agaggcactt 1741 tccaatgaga gaatttgtca gaaaaggaga ttttctgatt ctgaagaaag gcatacgaag 1801 aagcagtttt ctttggaaaa tgttcaggag ggggagattt tacacgatgc taagacattg 1861 gctggaaatg taatagctga cctatctgat tcttctgctg attctgaaaa tttaagtcca 1921 gatataaaag aaactactga ggaaatggaa tacaacatcc tcgtaaactt ttttaaaacc 1981 atgggctact cccaagaaat tgttgaaaag gtcattaagg tgtatggacc atctactgaa 2041 ccattattgc tcttagagga aattgaaaaa gaaaataaaa gattccaaga agacagagaa 2101 ttttcagctg gtacagtgta tccagagacc aacaaaacca aaaataaagg tgtttatagc 2161 agcacaaatg agcttacaac tgattccact ccaaagaaaa cacaggctca cacacagcaa 2221 aatatggtag aaaaattttc tcagttacca ttcaaagtgg aagctaaacc atgtacctca 2281 aattgtagaa ttaatacttt cagaacagtg ccaatagaac agaaacatga agtctggggt 2341 tcaaaccaga actacatttg taacacagac cctgaaactg atggcctttc accttctgtt 2401 gcctctccaa gtcccaaaga agtcaatttt gtttcaaggg gagcttcaag tcaccagccc 2461 agagttccac tttttcctga aaatggttta caccagcagc cagaaccctt gcttccaaat 2521 aatatgaaat ctgcctgtga aaaacgttta ggatgttgta gttctcctca ttctaagcca 2581 aattgctcaa ccctttctcc accaatgcca ctgccccagc tgttaccttc ggttactgat 2641 gcaaggtcgg caggaccttc tgatcatatt gattcctcag ttactggggt tcaaaggttt 2701 cgagatactc taaaaatacc ctacaagctg gaattaaaaa atgaaccagg gagaacggat 2761 ttgaaacaca ttgttataga tgggagcaat gttgcaatta cccatggtct gaaaaagttc 2821 ttttcttgtc gtggaattgc aattgcagtt gaatattttt ggaagcttgg caacagaaac 2881 atcactgtat ttgtccctca gtggagaaca aggcgtgatc ctaatgtcac agaacagcac 2941 ttcttaaccc agctccagga gctcggaata ttatctttaa ctcctgcccg gatggtcttt 3001 ggagaaagaa ttgcttctca taatgacagg tttctactac acttagcgga caaaactggt 3061 ggcataattg tgacaaatga taatttcaga gaatttgtga atgagtcagt ctcttggaga 3121 gaaattatta caaaaaggct gctgcagtac acgttcgtgg gggacatatt tatggttccc 3181 gatgatcctc tgggaagaag tggacctcga ttagaggagt ttcttcagaa ggaagtctgt 3241 cttagagata tgcagcccct actcagtgcc ctgccaaatg tgggcatgtt tgaccccagc 3301 ttcagagtcc ctggcaccca ggcagccagc accagccacc agcctccgac ccggattcag 3361 ggagcccctt caagccactg gctccctcag cagccccact tcccactgct gccagccctc 3421 cccagtctcc agcagaacct gcccatgcca gctcagagat cttctgcaga aaccaacgag 3481 ctgagggaag cccttctgaa gatcttccct gactcagagc aaagactgaa aatagaccag 3541 atcttggtgg cccacccata catgaaagat ctaaatgcgc tttctgccat ggtgttggat 3601 ctcgagaccg gtgactacaa agaccatgac ggtgattata aagatcatga catcgattac 3661 aaggatgacg atgacaagtg accggtaagt ttaaaccgct gatcagcctc gactgtgcct 3721 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt 3781 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg 3841 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac 3901 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc 3961 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 4021 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 4081 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 4141 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 4201 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 4261 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 4321 tcggtctatt cttttgattt ataagggatt ttggccattt cggcctattg gttaaaaaat 4381 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt 4441 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 4501 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 4561 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 4621 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 4681 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 4741 taggcttttg caaaaagctc ccgggatgac cgagtacaag cccacggtgc gcctcgccac 4801 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc 4861 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact 4921 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc 4981 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg 5041 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct 5101 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc 5161 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga 5221 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga 5281 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg 5341 catgacccgc aagcccggtg cctgattcga aatgaccgac caagcgacgc ccaacctgcc 5401 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 5461 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 5521 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 5581 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 5641 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 5701 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 5761 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 5821 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 5881 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 5941 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 6001 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 6061 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 6121 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 6181 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 6241 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 6301 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 6361 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 6421 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 6481 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 6541 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 6601 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac 6661 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 6721 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 6781 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 6841 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 6901 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 6961 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 7021 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 7081 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 7141 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 7201 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 7261 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 7321 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 7381 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 7441 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 7501 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 7561 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 7621 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 7681 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 7741 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 7801 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc //