LOCUS V038589 6526 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038589 VERSION V038589 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" regulatory 901..910 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 910..933 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 943..2046 /label="Fbxo28(NM_175127)" /note="Fbxo28(NM_175127)" /gene="Fbxo28" polyA_signal 2126..2350 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2396..2824 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2838..3167 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3234..4028 /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 4202..4323 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4372..4388) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4396..4412 /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(4420..4450) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4465..4486 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4774..5359) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5530..6390) /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(6391..6495) /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 gccaccatgg attacaagga tgacgacgat aagactaagc ttatggcggc ggcgtctgag 961 gagcggatgg ctgaggaagg aggcggcggc cacggcgacg gcggctcctg ttcggctgcc 1021 ggctccgctc agcgacagcc cccggcgccc ccgtcgcagg ccccgccgcc cgggtcccag 1081 gcacccgccg cgcccgcgct ggcaccggac cacctgcccc aaaacaacac actggtggcg 1141 ctgcccatcg tagccatcga gaacatcctc agctttatgt cctacgacga aattagccag 1201 ctccgcctgg tttgtaagag aatggacctg gtctgtcaga gaatgttgaa tcaggggttt 1261 ctgaaagtgg agaggttcca taacctgtgt cagaagcaag ttaaagcaca gctaccgagg 1321 agagagtcag aaagaagaaa ccattcttta gctcgtcatg cagacatcct tgccgctgtg 1381 gaaacgaggt tgtcactgtt gaatatgacc ttcatgaagt atgtggattc caacctctgt 1441 tgcttcatcc caggaaaggt gattgatgag atttaccgtg tactgagata tgtcaattct 1501 accagagccc ctcagcgagc tcacgaggtg ctgcaggagt tacgggatat ctcctccatg 1561 gcgatggagt actttgatga gaagattgtt ccaattttga agaggaaatt gccaggatca 1621 gatgtgtcag gaagactcat gggctctcct ccagttcccg ggccatccgc agccctgaca 1681 acaatgcagc tcttctccaa gcaaaacccg tccagacaag aggttaccaa actgcagcag 1741 caggtcagaa ccaacggcgc tggcgtgact gtcctcaggc gtgagatttc tgagcttcgc 1801 accaaagtgc aagagcagca gaaacagctt caagaccaag accagaaact gctcgagcag 1861 acccagatca taggcgaaca gaacgcacgg ctggcagagc tggaacgcaa gctccgggaa 1921 gtcatggaaa gtgcagtagg gacctcctca gggtctgggc agagtgagga gtcccctcgg 1981 aaacgacgga aggcaacgga agccatagac tctcttagga aatccaaacg acttcggaat 2041 aggaagtagg aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc 2101 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc 2161 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa 2221 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg 2281 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg 2341 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg 2401 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 2461 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 2521 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 2581 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 2641 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 2701 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 2761 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 2821 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 2881 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 2941 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 3001 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3061 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 3121 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 3181 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 3241 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 3301 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 3361 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 3421 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 3481 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 3541 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 3601 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 3661 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 3721 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 3781 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 3841 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 3901 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 3961 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 4021 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 4081 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 4141 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc 4201 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 4261 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 4321 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 4381 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 4441 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 4501 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 4561 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 4621 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 4681 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 4741 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 4801 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 4861 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 4921 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 4981 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 5041 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 5101 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 5161 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 5221 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 5281 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg 5341 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 5401 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 5461 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 5521 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 5581 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 5641 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 5701 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 5761 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 5821 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 5881 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 5941 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 6001 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 6061 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 6121 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 6181 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 6241 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 6301 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 6361 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 6421 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 6481 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc //