LOCUS V023811 5747 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023811 VERSION V023811 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 941..1237 /label="APOC3(NM_000040)" /note="APOC3(NM_000040)" /gene="APOC3" CDS 1244..1309 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1347..1571 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1617..2045 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2059..2388 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2455..3249 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 3423..3544 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3593..3609) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3617..3633 /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(3641..3671) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3686..3707 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(3995..4580) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4751..5611) /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(5612..5716) /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 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgcagcccc gggtactcct 961 tgttgttgcc ctcctggcgc tcctggcctc tgcccgagct tcagaggccg aggatgcctc 1021 ccttctcagc ttcatgcagg gttacatgaa gcacgccacc aagaccgcca aggatgcact 1081 gagcagcgtg caggagtccc aggtggccca gcaggccagg ggctgggtga ccgatggctt 1141 cagttccctg aaagactact ggagcaccgt taaggacaag ttctctgagt tctgggattt 1201 ggaccctgag gtcagaccaa cttcagccgt ggctgccctc gaggactaca aagaccatga 1261 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc 1321 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 1381 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 1441 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 1501 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 1561 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc 1621 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 1681 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 1741 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 1801 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 1861 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 1921 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 1981 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 2041 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 2101 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 2161 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 2221 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 2281 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta 2341 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga 2401 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt 2461 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 2521 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 2581 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac 2641 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 2701 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 2761 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 2821 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 2881 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 2941 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag 3001 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 3061 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 3121 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 3181 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 3241 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 3301 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 3361 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 3421 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 3481 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 3541 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 3601 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 3661 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 3721 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 3781 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 3841 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 3901 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 3961 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4021 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4081 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 4141 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 4201 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 4261 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 4321 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 4381 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 4441 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 4501 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc 4561 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 4621 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 4681 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 4741 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 4801 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 4861 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 4921 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 4981 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 5041 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 5101 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 5161 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 5221 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 5281 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 5341 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 5401 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 5461 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 5521 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 5581 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 5641 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 5701 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //