LOCUS V035302 5716 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035302 VERSION V035302 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 925..1467 /label="ATF3(NM_001674)" /note="ATF3(NM_001674)" /gene="ATF3" CDS 1480..1545 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1580..1804 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1850..2278 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2292..2621 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2633..3232 /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 3392..3513 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3562..3578) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3586..3602 /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(3610..3640) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3655..3676 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(3964..4549) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4720..5580) /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(5581..5685) /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 aagctttcta gagaattcgc caccatgatg cttcaacacc caggccaggt ctctgcctcg 961 gaagtgagtg cttctgccat cgtcccctgc ctgtcccctc ctgggtcact ggtgtttgag 1021 gattttgcta acctgacgcc ctttgtcaag gaagagctga ggtttgccat ccagaacaag 1081 cacctctgcc accggatgtc ctctgcgctg gaatcagtca ctgtcagcga cagacccctc 1141 ggggtgtcca tcacaaaagc cgaggtagcc cctgaagaag atgaaaggaa aaagaggcga 1201 cgagaaagaa ataagattgc agctgcaaag tgccgaaaca agaagaagga gaagacggag 1261 tgcctgcaga aagagtcgga gaagctggaa agtgtgaatg ctgaactgaa ggctcagatt 1321 gaggagctca agaacgagaa gcagcatttg atatacatgc tcaaccttca tcggcccacg 1381 tgtattgtcc gggctcagaa tgggaggact ccagaagatg agagaaacct ctttatccaa 1441 cagataaaag aaggaacatt gcagagcctc gagaccggtg actacaaaga ccatgacggt 1501 gattataaag atcatgacat cgattacaag gatgacgatg acaagtgacc ggtaagttta 1561 aaccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc 1621 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag 1681 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag 1741 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct 1801 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt 1861 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 1921 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 1981 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 2041 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga 2101 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 2161 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg 2221 gccatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa 2281 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa 2341 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc 2401 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc 2461 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 2521 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga 2581 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggatgaccga 2641 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct 2701 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat 2761 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 2821 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 2881 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 2941 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 3001 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 3061 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 3121 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 3181 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gattcgaaat 3241 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta 3301 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 3361 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta 3421 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 3481 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag 3541 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 3601 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 3661 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 3721 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 3781 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 3841 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 3901 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 3961 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 4021 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 4081 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 4141 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 4201 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 4261 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 4321 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 4381 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 4441 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 4501 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 4561 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 4621 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 4681 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 4741 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 4801 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 4861 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 4921 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 4981 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 5041 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 5101 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 5161 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 5221 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 5281 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 5341 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 5401 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 5461 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 5521 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 5581 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 5641 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 5701 agtgccacct gacgtc //