LOCUS V033684 7384 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033684 VERSION V033684 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 919..3135 /label="SUZ12(NM_015355)" /note="SUZ12(NM_015355)" /gene="SUZ12" CDS 3148..3213 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3248..3472 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3518..3946 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3960..4289 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4301..4900 /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 5060..5181 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5230..5246) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5254..5270 /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(5278..5308) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5323..5344 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5632..6217) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6388..7248) /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(7249..7353) /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 gagccaccat ggcgcctcag aagcacggcg gtgggggagg gggcggctcg 961 gggcccagcg cggggtccgg gggaggcggc ttcgggggtt cggcggcggt ggcggcggcg 1021 acggcttcgg gcggcaaatc cggcggcggg agctgtggag ggggtggcag ttactcggcc 1081 tcctcctcct cctccgcggc ggcagcggcg ggggctgcgg tgttaccggt gaagaagccg 1141 aaaatggagc acgtccaggc tgaccacgag cttttcctcc aggcctttga gaagccaaca 1201 cagatctata gatttcttcg aactcggaat ctcatagcac caatattttt gcacagaact 1261 cttacttaca tgtctcatcg aaactccaga acaaacatca aaaggaaaac atttaaagtt 1321 gatgatatgt tatcaaaagt agagaaaatg aaaggagagc aagaatctca tagcttgtca 1381 gctcatttgc agcttacgtt tactggtttc ttccacaaaa atgataagcc atcaccaaac 1441 tcagaaaatg aacaaaattc tgttaccctg gaagtcctgc ttgtgaaagt ttgccacaaa 1501 aaaagaaagg atgtaagttg tccaataagg caagttccca caggtaaaaa gcaggtgcct 1561 ttgaatcctg acctcaatca aacaaaaccc ggaaatttcc cgtcccttgc agtttccagt 1621 aatgaatttg aacctagtaa cagccatatg gtgaagtctt actcgttgct atttagagtg 1681 actcgtccag gaagaagaga gtttaatgga atgattaatg gagaaaccaa tgaaaatatt 1741 gatgtcaatg aagagcttcc agccagaaga aaacgaaatc gtgaggatgg ggaaaagaca 1801 tttgttgcac aaatgacagt atttgataaa aacaggcgct tacagctttt agatggggaa 1861 tatgaagtag ccatgcagga aatggaagaa tgtccaataa gcaagaaaag agcaacatgg 1921 gagactattc ttgatgggaa gaggctgcct ccattcgaaa cattttctca gggacctacg 1981 ttgcagttca ctcttcgttg gacaggagag accaatgata aatctacggc tcctattgcc 2041 aaacctcttg ccactagaaa ttcagagagt ctccatcagg aaaacaagcc tggttcagtt 2101 aaacctactc aaactattgc tgttaaagaa tcattgacta cagatctaca aacaagaaaa 2161 gaaaaggata ctccaaatga aaaccgacaa aaattaagaa tattttatca gtttctctat 2221 aacaacaata caaggcaaca aactgaagca agagatgacc tgcattgccc ttggtgtact 2281 ctgaactgcc gcaaacttta tagtttactc aagcatctta aactctgcca tagcagattt 2341 atcttcaact atgtttatca tccaaaaggt gctaggatag atgtttctat caatgagtgt 2401 tatgatggct cctatgcagg aaatcctcag gatattcatc gccaacctgg atttgctttt 2461 agtcgcaacg gaccagttaa gagaacacct atcacacata ttcttgtgtg caggccaaaa 2521 cgaacaaaag caagcatgtc tgaatttctt gaatctgaag atggggaagt agaacagcaa 2581 agaacatata gtagtggcca caatcgtctg tatttccata gtgatacctg cttacctctc 2641 cgtccacaag aaatggaagt agatagtgaa gatgaaaagg atcctgaatg gctaagagaa 2701 aaaaccatta cacaaattga agagttttct gatgttaatg aaggagagaa agaagtgatg 2761 aaactctgga atctccatgt catgaagcat gggtttattg ctgacaatca aatgaatcat 2821 gcctgtatgc tgtttgtaga aaattatgga cagaaaataa ttaagaagaa tttatgtcga 2881 aacttcatgc ttcatctagt cagcatgcat gactttaatc ttattagcat aatgtcaata 2941 gataaagctg ttaccaagct ccgtgaaatg cagcaaaaat tagaaaaggg ggaatctgct 3001 tcccctgcaa acgaagaaat aactgaagaa caaaatggga cagcaaatgg atttagtgaa 3061 attaactcaa aagagaaagc tttggaaaca gatagtgtct caggggtttc aaaacagagc 3121 aaaaaacaaa aactcctcga gaccggtgac tacaaagacc atgacggtga ttataaagat 3181 catgacatcg attacaagga tgacgatgac aagtgaccgg taagtttaaa ccgctgatca 3241 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 3301 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 3361 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 3421 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag 3481 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta 3541 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 3601 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 3661 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 3721 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt 3781 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 3841 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggc catttcggcc 3901 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg 3961 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 4021 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 4081 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 4141 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 4201 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag 4261 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg atgaccgagt acaagcccac 4321 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt 4381 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac 4441 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc 4501 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt 4561 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca 4621 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac 4681 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg 4741 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa 4801 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg 4861 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ttcgaaatga ccgaccaagc 4921 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 4981 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 5041 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa 5101 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 5161 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc 5221 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 5281 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 5341 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 5401 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 5461 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 5521 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 5581 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 5641 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 5701 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 5761 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 5821 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 5881 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 5941 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 6001 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 6061 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 6121 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg 6181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 6241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 6301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 6361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 6421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 6481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 6541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 6601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 6661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 6721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 6781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 6841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 6901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 6961 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 7021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 7081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 7141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 7201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 7261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 7321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 7381 cgtc //