LOCUS V023239 6248 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023239 VERSION V023239 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..1738 /label="RASD2(NM_014310)" /note="RASD2(NM_014310)" /gene="RASD2" CDS 1745..1810 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1848..2072 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2118..2546 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2560..2889 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2956..3750 /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 3924..4045 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4094..4110) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4118..4134 /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(4142..4172) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4187..4208 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4496..5081) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5252..6112) /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(6113..6217) /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 atgatgaaga ctttgtccag 961 cgggaactgc acgctcagtg tgcccgccaa aaactcatac cgcatggtgg tgctgggtgc 1021 ctctcgggtg ggcaagagct ccatcgtgtc tcgcttcctc aatggccgct ttgaggacca 1081 gtacacaccc accatcgagg acttccaccg taaggtatac aacatccgcg gcgacatgta 1141 ccagctcgac atcctggata cctctggcaa ccaccccttc cccgccatgc gcaggctgtc 1201 catcctcaca ggggatgtct tcatcctggt gttcagcctg gataaccggg agtccttcga 1261 tgaggtcaag cgccttcaga agcagatcct ggaggtcaag tcctgcctga agaacaagac 1321 caaggaggcg gcggagctgc ccatggtcat ctgtggcaac aagaacgacc acggcgagct 1381 gtgccgccag gtgcccacca ccgaggccga gctgctggtg tcgggcgacg agaactgcgc 1441 ctacttcgag gtgtcggcca agaagaacac caacgtggac gagatgttct acgtgctctt 1501 cagcatggcc aagctgccac acgagatgag ccccgccctg catcgcaaga tctccgtgca 1561 gtacggtgac gccttccacc ccaggccctt ctgcatgcgc cgcgtcaagg agatggacgc 1621 ctatggcatg gtctcgccct tcgcccgccg ccccagcgtc aacagtgacc tcaagtacat 1681 caaggccaag gtccttcggg aaggccaggc ccgtgagagg gacaagtgca ccatccagct 1741 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga 1801 cgatgacaag tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag 1861 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac 1921 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 1981 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag 2041 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg 2101 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 2161 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 2221 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 2281 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 2341 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 2401 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 2461 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 2521 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga 2581 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 2641 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 2701 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 2761 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 2821 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 2881 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga 2941 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 3001 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 3061 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 3121 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 3181 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 3241 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 3301 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 3361 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 3421 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 3481 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 3541 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 3601 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 3661 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 3721 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc 3781 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 3841 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 3901 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa 3961 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 4021 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct 4081 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 4141 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 4201 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 4261 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 4321 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 4381 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 4441 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 4501 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 4561 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 4621 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 4681 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 4741 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 4801 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 4861 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 4921 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 4981 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg 5041 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 5101 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 5161 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 5221 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 5281 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 5341 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 5401 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 5461 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 5521 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 5581 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 5641 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 5701 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 5761 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 5821 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 5881 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 5941 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 6001 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 6061 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 6121 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 6181 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 6241 ctgacgtc //