LOCUS V038185 6082 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038185 VERSION V038185 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..1572 /label="RAB29(NM_003929)" /note="RAB29(NM_003929)" /gene="RAB29" CDS 1579..1644 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1682..1906 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1952..2380 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2394..2723 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2790..3584 /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 3758..3879 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3928..3944) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3952..3968 /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(3976..4006) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4021..4042 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4330..4915) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5086..5946) /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(5947..6051) /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 accatgggca gccgcgacca cctgttcaaa gtgctggtgg tgggggacgc cgcagtgggc 1021 aagacgtcgc tggtgcagcg atattcccag gacagcttca gcaaacacta caagtccacg 1081 gtgggagtgg attttgctct gaaggttctc cagtggtctg actacgagat agtgcggctt 1141 cagctgtggg atattgcagg gcaggagcgc ttcacctcta tgacacgatt gtattatcgg 1201 gatgcctctg cctgtgttat tatgtttgac gttaccaatg ccactacctt cagcaacagc 1261 cagaggtgga aacaggacct agacagcaag ctcacactac ccaatggaga gccggtgccc 1321 tgcctgctct tggccaacaa gtgtgatctg tccccttggg cagtgagccg ggaccagatt 1381 gaccggttca gtaaagagaa cggtttcaca ggttggacag aaacatcagt caaggagaac 1441 aaaaatatta atgaggctat gagagtcctc attgaaaaga tgatgagaaa ttccacagaa 1501 gatatcatgt ctttgtccac ccaaggggac tacatcaatc tacaaaccaa gtcctccagc 1561 tggtcctgct gcctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc 1621 gactacaagg atgacgatga caagtagtga gggcccgttt aaacccgctg atcagcctcg 1681 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 1741 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 1801 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 1861 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa 1921 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 1981 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 2041 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 2101 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 2161 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 2221 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 2281 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 2341 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 2401 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 2461 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 2521 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 2581 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 2641 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 2701 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 2761 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 2821 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 2881 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 2941 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 3001 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 3061 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 3121 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 3181 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 3241 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 3301 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 3361 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 3421 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 3481 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 3541 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 3601 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc 3661 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 3721 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa 3781 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 3841 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac 3901 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 3961 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 4021 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 4081 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 4141 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 4201 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 4261 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 4321 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 4381 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 4441 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 4501 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 4561 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 4621 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 4681 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 4741 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 4801 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 4861 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 4921 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 4981 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 5041 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 5101 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 5161 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 5221 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 5281 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 5341 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 5401 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 5461 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 5521 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 5581 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 5641 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 5701 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 5761 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 5821 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 5881 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 5941 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 6001 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 6061 ccgaaaagtg ccacctgacg tc //