LOCUS V024868 6401 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024868 VERSION V024868 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..1891 /label="APOE(NM_000041)" /note="APOE(NM_000041)" /gene="APOE" CDS 1898..1963 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2001..2225 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2271..2699 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2713..3042 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3109..3903 /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 4077..4198 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4247..4263) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4271..4287 /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(4295..4325) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4340..4361 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4649..5234) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5405..6265) /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(6266..6370) /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 atgaaggttc tgtgggctgc 961 gttgctggtc acattcctgg caggatgcca ggccaaggtg gagcaagcgg tggagacaga 1021 gccggagccc gagctgcgcc agcagaccga gtggcagagc ggccagcgct gggaactggc 1081 actgggtcgc ttttgggatt acctgcgctg ggtgcagaca ctgtctgagc aggtgcagga 1141 ggagctgctc agctcccagg tcacccagga actgagggcg ctgatggacg agaccatgaa 1201 ggagttgaag gcctacaaat cggaactgga ggaacaactg accccggtgg cggaggagac 1261 gcgggcacgg ctgtccaagg agctgcaggc ggcgcaggcc cggctgggcg cggacatgga 1321 ggacgtgtgc ggccgcctgg tgcagtaccg cggcgaggtg caggccatgc tcggccagag 1381 caccgaggag ctgcgggtgc gcctcgcctc ccacctgcgc aagctgcgta agcggctcct 1441 ccgcgatgcc gatgacctgc agaagcgcct ggcagtgtac caggccgggg cccgcgaggg 1501 cgccgagcgc ggcctcagcg ccatccgcga gcgcctgggg cccctggtgg aacagggccg 1561 cgtgcgggcc gccactgtgg gctccctggc cggccagccg ctacaggagc gggcccaggc 1621 ctggggcgag cggctgcgcg cgcggatgga ggagatgggc agccggaccc gcgaccgcct 1681 ggacgaggtg aaggagcagg tggcggaggt gcgcgccaag ctggaggagc aggcccagca 1741 gatacgcctg caggccgagg ccttccaggc ccgcctcaag agctggttcg agcccctggt 1801 ggaagacatg cagcgccagt gggccgggct ggtggagaag gtgcaggctg ccgtgggcac 1861 cagcgccgcc cctgtgccca gcgacaatca cctcgaggac tacaaagacc atgacggtga 1921 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta 1981 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc 2041 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga 2101 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca 2161 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc 2221 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg 2281 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 2341 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 2401 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 2461 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg 2521 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 2581 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt 2641 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta 2701 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 2761 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 2821 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 2881 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 2941 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag 3001 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt 3061 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa 3121 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg 3181 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc 3241 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca 3301 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc 3361 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca 3421 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat 3481 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca 3541 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg 3601 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc 3661 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct 3721 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct 3781 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac 3841 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc 3901 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag 3961 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 4021 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact 4081 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 4141 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 4201 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc 4261 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 4321 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc 4381 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg 4441 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct 4501 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 4561 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 4621 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 4681 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 4741 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 4801 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 4861 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 4921 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 4981 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 5041 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 5101 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 5161 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa 5221 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 5281 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 5341 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 5401 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 5461 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 5521 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 5581 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 5641 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 5701 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 5761 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 5821 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 5881 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 5941 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 6001 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 6061 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 6121 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 6181 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 6241 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 6301 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 6361 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c //