LOCUS V032538 6116 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032538 VERSION V032538 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..1606 /label="CHMP4A(NM_014169)" /note="CHMP4A(NM_014169)" /gene="CHMP4A" CDS 1613..1678 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1716..1940 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1986..2414 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2428..2757 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2824..3618 /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 3792..3913 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3962..3978) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3986..4002 /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(4010..4040) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4055..4076 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4364..4949) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5120..5980) /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(5981..6085) /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 atgagtggtc tcggcaggct 961 cttcgggaag gggaagaagg agaaagggcc aacccctgaa gaagcaatac agaaactgaa 1021 ggagacagag aagatactga tcaagaaaca ggaatttttg gagcagaaga ttcaacagga 1081 gctacaaaca gccaagaagt atgggaccaa gaataagaga gctgccctac aggctttgcg 1141 gaggaagaaa agattcgaac agcagctggc acaaactgac gggacattat ccaccctgga 1201 gtttcagcgt gaggccattg agaatgccac taccaatgca gaagtccttc gtaccatgga 1261 gcttgctgcc caaagcatga agaaggccta ccaggacatg gacattgaca aggtagatga 1321 actgatgact gacatcacgg aacaacagga ggtggcccag cagatctcag atgccatttc 1381 tcggcctatg ggctttggag atgatgtgga tgaggatgaa ctgctggagg agctagagga 1441 gctggagcag gaggaattgg cccaggagtt gttaaatgtg ggcgacaagg aagaagaacc 1501 ctcagtcaaa ttgcctagtg taccttctac tcatctgccg gcagggccag ctcccaaagt 1561 ggatgaagat gaagaagcac taaagcagtt ggctgagtgg gtatccctcg aggactacaa 1621 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 1681 gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 1741 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 1801 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 1861 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 1921 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta 1981 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 2041 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 2101 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 2161 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 2221 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 2281 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 2341 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 2401 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 2461 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 2521 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 2581 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 2641 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 2701 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 2761 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 2821 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 2881 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 2941 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 3001 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 3061 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 3121 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 3181 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 3241 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 3301 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 3361 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 3421 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 3481 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 3541 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 3601 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 3661 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 3721 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 3781 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 3841 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 3901 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 3961 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 4021 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 4081 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 4141 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 4201 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 4261 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 4321 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 4381 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 4441 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 4501 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 4561 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 4621 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 4681 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 4741 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 4801 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 4861 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc 4921 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 4981 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 5041 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 5101 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 5161 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 5221 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 5281 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 5341 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 5401 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 5461 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 5521 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 5581 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 5641 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 5701 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 5761 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 5821 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 5881 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 5941 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 6001 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 6061 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc //