LOCUS V028311 6326 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028311 VERSION V028311 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..1816 /label="FBXO16(NM_172366)" /note="FBXO16(NM_172366)" /gene="FBXO16" CDS 1823..1888 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1926..2150 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2196..2624 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2638..2967 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3034..3828 /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 4002..4123 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4172..4188) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4196..4212 /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(4220..4250) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4265..4286 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4574..5159) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5330..6190) /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(6191..6295) /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 atgatggcat ttgcacctcc 961 aaaaaacaca gatggtccca aaatgcagac aaagatgagc acctggacac ccctaaacca 1021 tcagctattg aatgaccggg tatttgaaga aagaagagcc ctgcttggca aatggtttga 1081 caaatggaca gactctcaaa gaagaagaat cctcacaggc ctgttggagc gctgctcgct 1141 gtcccagcaa aagttctgct gtcgaaagct tcaagagaaa attccagcag aagccctgga 1201 ctttacaacc aagcttccaa gggtgttatc tttatacatc ttttctttcc tggaccctcg 1261 gagcctttgt cgttgtgcac aggtgtgctg gcattggaag aaccttgctg agctggacca 1321 gctctggatg ctgaaatgtt tacggtttaa ctggtacatc aatttctctc caactccctt 1381 tgagcagggg atctggaaga agcactatat tcaaatggtg aaagaacttc atattaccaa 1441 gcctaagaca cccccaaagg atggatttgt aatcgctgac gttcaactag ttacaagcaa 1501 ttctccagag gaaaaacagt cccctttatc agcttttcgg tcctcttcct ctttaagaaa 1561 gaagaataac tcaggggaga aagcacttcc accctggcga tcttctgata agcacccaac 1621 agatatcatt cgttttaatt acctagacaa ccgtgacccc atggagactg tccagcaagg 1681 aagaagaaaa agaaaccaaa tgaccccaga cttcagccga cagtcacatg ataagaaaaa 1741 taaattgcag gacagaacta ggctaagaaa agcacaatca atgatgtcga ggagaaatcc 1801 cttcccacta tgtcccctcg aggactacaa agaccatgac ggtgattata aagatcatga 1861 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc 1921 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 1981 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 2041 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 2101 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 2161 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 2221 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 2281 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 2341 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 2401 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 2461 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 2521 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 2581 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 2641 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 2701 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 2761 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 2821 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 2881 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 2941 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 3001 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 3061 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 3121 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 3181 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 3241 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 3301 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 3361 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 3421 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 3481 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 3541 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 3601 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 3661 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 3721 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 3781 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 3841 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 3901 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 3961 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 4021 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 4081 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 4141 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 4201 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 4261 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 4321 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 4381 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 4441 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 4501 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4561 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 4621 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 4681 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 4741 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 4801 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 4861 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 4921 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 4981 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 5041 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 5101 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 5161 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 5221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 5281 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 5341 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 5401 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 5461 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 5521 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 5581 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 5641 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 5701 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 5761 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 5821 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 5881 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 5941 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 6001 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 6061 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 6121 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 6181 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 6241 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 6301 ttccccgaaa agtgccacct gacgtc //