LOCUS V045118 6064 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045118 VERSION V045118 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..1815 /label="Emc2(NM_025736)" /note="Emc2(NM_025736)" /gene="Emc2" CDS 1828..1893 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1928..2152 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2198..2626 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2640..2969 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2981..3580 /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 3740..3861 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3910..3926) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3934..3950 /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(3958..3988) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4003..4024 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4312..4897) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5068..5928) /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(5929..6033) /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 caccatggcg aaggtcactg agcgttacga tgtgacttgg 961 gaagaaatgc gagataaaat gagaaaatgg cgagaagaaa actcaagaaa tagtgaacaa 1021 attatggaag ttggagaaga attaattaat gattatgctt ctaagcttgg ggatgacatt 1081 tggatcatat atgagcaggt catgattgca gccctagact atggtcggga tgacttggca 1141 ttgttttgtc ttcaggaatt aagaagacaa ttccctggta gtcacagagt taagcggtta 1201 acaggcatgc gatttgaagc tatggaaaga tatgacgatg ctatacaact gtatgatcgg 1261 attttgcaag aagatccaac taacactgct gccagaaagc gtaagattgc cattcgaaaa 1321 gcccagggga aaaccgtgga ggccatccga gagctgaatg agtatctgga gcagtttgtt 1381 ggagaccaag aagcctggca tgaacttgca gaactttata tcaatgaaca cgactatgcc 1441 aaagcagctt tttgcttaga ggagctgatg atgacaaatc cacataacca cttatactgt 1501 cagcagtatg cagaagtcaa atacacccaa ggtggacttg aaaacctcga gctttcaaga 1561 aagtattttg cacaggcact gaaactcaac aacaggaaca tgagagctct gtttgggctc 1621 tacatgtctg caagtcatat tgcttctaat ccaaaagcaa gtgcaaaaat gaaaaaagac 1681 aacatcaaat atgctagttg ggccgctaat caaataaaca gggcttatca gtttgcaggt 1741 cgaagtaaga aggaaaccaa atactctctt aaggcagttg aagacatgtt ggagacattg 1801 cagatcactc agtctctcga gaccggtgac tacaaagacc atgacggtga ttataaagat 1861 catgacatcg attacaagga tgacgatgac aagtgaccgg taagtttaaa ccgctgatca 1921 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 1981 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 2041 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 2101 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag 2161 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta 2221 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 2281 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 2341 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 2401 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt 2461 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 2521 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggc catttcggcc 2581 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg 2641 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 2701 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 2761 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 2821 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 2881 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag 2941 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg atgaccgagt acaagcccac 3001 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt 3061 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac 3121 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc 3181 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt 3241 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca 3301 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac 3361 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg 3421 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa 3481 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg 3541 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ttcgaaatga ccgaccaagc 3601 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 3661 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 3721 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa 3781 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 3841 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc 3901 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 3961 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 4021 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 4081 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 4141 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 4201 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 4261 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 4321 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 4381 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 4441 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 4501 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 4561 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 4621 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 4681 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 4741 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 4801 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg 4861 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 4921 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 4981 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 5041 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 5101 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 5161 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 5221 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 5281 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 5341 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 5401 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 5461 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 5521 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 5581 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 5641 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 5701 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 5761 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 5821 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 5881 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 5941 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 6001 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 6061 cgtc //