LOCUS V034379 6673 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V034379 VERSION V034379 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..2424 /label="PLAG1(NM_002655)" /note="PLAG1(NM_002655)" /gene="PLAG1" CDS 2437..2502 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2537..2761 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2807..3235 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3249..3578 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3590..4189 /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 4349..4470 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4519..4535) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4543..4559 /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(4567..4597) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4612..4633 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4921..5506) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5677..6537) /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(6538..6642) /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 caccatggcc actgtcattc ctggtgattt gtcagaagta 961 agagataccc agaaagtccc ttcagggaaa cgtaagcgtg gtgaaaccaa accaagaaaa 1021 aactttcctt gccaactgtg tgacaaggcc tttaacagtg ttgagaaatt aaaggttcac 1081 tcctactctc acacaggaga gaggccctac aagtgcatac aacaagactg caccaaggcc 1141 tttgtttcta agtacaaatt acaaaggcac atggctactc attctcctga gaaaacccac 1201 aagtgtaatt attgtgagaa aatgtttcac cggaaagatc atctgaagaa tcacctccat 1261 acacacgacc ctaacaaaga gacgtttaag tgcgaagaat gtggcaagaa ctacaatacc 1321 aagcttggat ttaaacgtca cttggccttg catgccgcaa caagtggtga cctcacctgt 1381 aaggtatgtt tgcaaacttt tgaaagcacg ggagtgcttc tggagcacct taaatctcat 1441 gcaggcaagt cgtctggtgg ggttaaagaa aaaaagcacc agtgcgaaca ttgtgatcgc 1501 cggttctaca cccgaaagga tgtccggaga cacatggtgg tgcacactgg aagaaaggac 1561 ttcctctgtc agtattgtgc acagagattt gggcgaaagg atcacctgac tcgacatatg 1621 aagaagagtc acaatcaaga gcttctgaag gtcaaaacag aaccagtgga tttccttgac 1681 ccatttacct gcaatgtgtc tgtgcctata aaagacgagc tccttccggt gatgtcctta 1741 ccttccagtg aactgttatc aaagccattc acaaacactt tgcagttaaa cctctacaac 1801 actccatttc agtccatgca gagctcggga tctgcccacc aaatgatcac aactttacct 1861 ttgggaatga catgcccaat agatatggac actgttcatc cctctcacca cctttctttc 1921 aaatatccgt tcagttctac ctcatatgca atttctattc ctgaaaaaga acagccatta 1981 aagggggaaa ttgagagtta cctgatggag ttacaaggtg gcgtgccctc ttcatcccaa 2041 gattctcaag catcgtcatc atctaagcta gggttggatc ctcagattgg gtccctagat 2101 gatggtgcag gagacctctc cctatccaaa agctctatct ccatcagtga ccccctaaac 2161 acaccagcat tggatttttc tcagttgttt aatttcatac ctttaaatgg tcctccctat 2221 aatcctctat cagtggggag ccttggaatg agctattccc aggaagaagc acattcttct 2281 gtttcccagc tccccccaca aacacaggat cttcaggatc ctgcaaacac tatagggctt 2341 gggtctctgc actcactgtc agcagctttc accagcagtt taagcacaag taccaccctc 2401 ccacgtttcc atcaagcttt tcagctcgag accggtgact acaaagacca tgacggtgat 2461 tataaagatc atgacatcga ttacaaggat gacgatgaca agtgaccggt aagtttaaac 2521 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 2581 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 2641 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 2701 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 2761 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 2821 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 2881 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 2941 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 3001 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 3061 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 3121 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttggcc 3181 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 3241 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 3301 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 3361 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3421 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 3481 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 3541 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga tgaccgagta 3601 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc 3661 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga 3721 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt 3781 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc 3841 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc 3901 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt 3961 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt 4021 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc 4081 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt 4141 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgat tcgaaatgac 4201 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga 4261 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga 4321 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa 4381 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 4441 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta 4501 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 4561 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 4621 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 4681 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 4741 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 4801 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 4861 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 4921 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 4981 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 5041 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 5101 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 5161 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 5221 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 5281 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 5341 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 5401 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt 5461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 5521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 5581 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 5641 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 5701 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 5761 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 5821 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 5881 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 5941 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 6001 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 6061 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 6121 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 6181 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 6241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 6301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 6361 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 6421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 6481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 6541 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 6601 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 6661 gccacctgac gtc //