LOCUS V039959 8755 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039959 VERSION V039959 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 964..4245 /label="AP3B1(NM_003664)" /note="AP3B1(NM_003664)" /gene="AP3B1" CDS 4252..4317 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 4355..4579 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4625..5053 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5067..5396 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5463..6257 /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 6431..6552 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6601..6617) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6625..6641 /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(6649..6679) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6694..6715 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7003..7588) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7759..8619) /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(8620..8724) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatgtcca gcaatagttt tccttacaat gagcagtccg gaggagggga ggcgacggag 1021 ctgggtcagg aggcgacctc aaccatttcc ccctcggggg ccttcggcct ctttagcagc 1081 gatttgaaga agaatgaaga tctaaagcaa atgttagaga gcaacaaaga ttctgctaaa 1141 ctggatgcta tgaagcggat tgttgggatg attgcaaaag ggaaaaatgc atctgaactg 1201 tttcctgctg ttgtgaagaa tgtggccagt aaaaatattg agatcaagaa gttggtatat 1261 gtttacctgg ttcgatatgc tgaagaacag caggatcttg cactcctgtc cataagcact 1321 tttcagcgag ctctgaagga cccaaaccaa ctaattcgtg caagcgcttt gagagttctg 1381 tcaagtatta gagtgccaat tattgtacct atcatgatgc ttgctattaa ggaagcttct 1441 gctgacttat caccatatgt taggaagaat gcagcccatg caatacaaaa attatacagc 1501 cttgatccag agcagaagga aatgttaatt gaagtaattg aaaaacttct gaaagataaa 1561 agcacattgg tagctggcag tgttgtgatg gcttttgaag aagtatgccc ggacagaata 1621 gatctgattc ataaaaatta ccgcaagcta tgtaacttac tagtggatgt tgaagagtgg 1681 gggcaggttg tcataatcca catgctaact cgatatgctc ggacacagtt tgtcagccct 1741 tggaaagagg gtgatgaatt agaagacaat ggaaagaatt tctacgaatc tgatgatgat 1801 cagaaggaaa agactgacaa aaagaagaag ccgtatacta tggatccaga tcatagactc 1861 ttaattagaa atacaaagcc tttgcttcag agcaggaatg ctgcggtggt tatggcagtt 1921 gctcagctgt attggcacat atcaccaaaa tctgaagctg gcataatttc taaatcacta 1981 gtgcgtttac ttcgtagcaa tagggaggtg cagtatattg tcctacaaaa tatagcaact 2041 atgtcaattc aaagaaaggg gatgtttgaa ccttatctga agagtttcta tgttaggtca 2101 actgatccaa ctatgatcaa gacactgaag cttgaaattt tgacaaactt ggcaaatgaa 2161 gccaacatat caactcttct tcgagaattt cagacctatg tgaaaagcca ggataaacaa 2221 tttgcagcag ccactattca gactataggc agatgtgcaa ccaacatctt ggaagtcact 2281 gacacgtgcc tcaatggctt ggtctgtctg ctgtccaaca gggatgaaat agttgttgct 2341 gaaagtgtgg ttgttataaa gaaattactg caaatgcaac ctgcacaaca tggtgaaatt 2401 attaaacata tggccaaact cctggacagt atcactgttc ctgttgctag agcaagtatt 2461 ctttggctaa ttggagaaaa ctgtgaacga gttcctaaaa ttgcccctga tgttttgagg 2521 aagatggcta aaagcttcac tagtgaagat gatctggtaa aactgcagat attaaatctg 2581 ggagcaaaat tgtatttaac caactccaaa cagacaaaat tgcttaccca gtacatatta 2641 aatctcggca agtatgatca aaactacgac atcagagacc gtacaagatt tattaggcag 2701 cttattgttc cgaatgtaaa gagtggagct ttaagtaaat atgccaaaaa aatattccta 2761 gcacaaaagc ctgcaccact gcttgagtct ccttttaaag atagagatca tttccagctt 2821 ggcaccttat ctcatactct caacattaaa gctactgggt acctggaatt atctaattgg 2881 ccagaggtgg cgcccgaccc atcagttcga aatgtagaag taatagagtt ggcaaaagaa 2941 tggaccccag caggaaaagc aaagcaagag aattctgcta agaagtttta ttctgaatct 3001 gaggaagagg aggactcttc tgatagtagc agtgacagtg agagtgaatc tggaagtgaa 3061 agtggagaac aaggcgaaag tggggaggaa ggagacagca atgaggacag cagtgaggac 3121 tcctccagtg agcaggacag tgagagtgga cgggagtcag gcctagaaaa caaaagaaca 3181 gccaagagga actcaaaagc caaaggaaaa agtgattctg aagatgggga gaaggaaaat 3241 gaaaaatcta aaacttcaga ttcttcaaat gacgaatcta gttcaataga agacagttct 3301 tccgattctg aatcagagtc agaacctgaa agtgaatctg aatccagaag agtcactaag 3361 gagaaagaaa agaaaacaaa gcaagataga actcctctta ccaaagatgt ttcacttcta 3421 gatctggatg attttaaccc agtatccact ccagttgcac ttcccacacc agctctttct 3481 ccaagtttga tggctgatct tgaaggttta cacttgtcaa cttcctcttc agtcatcagt 3541 gtcagtactc ctgcatttgt accaacgaaa actcacgtgc tgcttcatcg aatgagtgga 3601 aaaggactag ctgcccatta tttctttcca agacagcctt gcatttttgg tgataagatg 3661 gtctctatac aaataacact gaataacact actgatcgaa agatagaaaa tatccacata 3721 ggggaaaaaa aacttcctat aggcatgaaa atgcatgttt ttaatccaat agactctctt 3781 gagcctgagg gatccattac agtttcaatg ggtattgact tttgtgattc tactcagact 3841 gccagtttcc agttgtgtac caaggatgat tgcttcaatg ttaatattca gccacctgtt 3901 ggagaactgc ttttacctgt ggccatgtca gagaaagatt ttaagaaaga gcaaggagtg 3961 ctaacaggaa tgaatgaaac ttctgctgta atcattgctg caccacagaa tttcactccc 4021 tctgtgatct ttcagaaggt tgtaaatgta gccaatgtag gtgcagtccc ttctggccag 4081 gataatatac acaggtttgc agctaaaact gtgcacagtg ggtcattgat gctagtcaca 4141 gtggaactga aggaaggctc tacagcccag cttatcataa acactgagaa aactgtgatt 4201 ggctctgttc tgctgcggga actgaagcct gtcctgtctc aggggctcga ggactacaaa 4261 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 4321 tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct 4381 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 4441 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 4501 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 4561 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat 4621 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 4681 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 4741 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 4801 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 4861 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 4921 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 4981 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 5041 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 5101 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 5161 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 5221 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 5281 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 5341 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 5401 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 5461 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 5521 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 5581 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 5641 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 5701 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 5761 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 5821 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 5881 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 5941 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 6001 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 6061 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 6121 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 6181 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 6241 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 6301 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 6361 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 6421 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 6481 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 6541 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg 6601 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6661 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6721 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6781 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6841 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6901 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6961 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7021 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7081 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7141 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7201 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7261 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7321 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7381 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7441 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7501 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca 7561 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 7621 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 7681 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 7741 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 7801 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 7861 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 7921 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 7981 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 8041 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 8101 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 8161 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 8221 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 8281 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 8341 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 8401 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 8461 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 8521 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 8581 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 8641 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 8701 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //