LOCUS V031488 7235 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031488 VERSION V031488 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 683..1915 /label="EIF4A3(NM_014740)" /note="EIF4A3(NM_014740)" /gene="EIF4A3" CDS 1922..1987 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2011..2599 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2665..2713 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2915..3343 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3357..3686 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3753..4547 /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 4723..4844 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4893..4909) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4917..4933 /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(4941..4971) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4986..5007 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5295..5883) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6054..6914) /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(6915..7019) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccgcca ccatggcgac cacggccacg atggcgacct cgggctcggc 721 gcgaaagcgg ctgctcaaag aggaagacat gactaaagtg gaattcgaga ccagcgagga 781 ggtggatgtg acccccacgt tcgacaccat gggcctgcgg gaggacctgc tgcggggcat 841 ctacgcttac ggttttgaaa aaccatcagc aatccagcaa cgagcaatca agcagatcat 901 caaagggaga gatgtcatcg cacagtctca gtccggcaca ggaaaaacag ccaccttcag 961 tatctcagtc ctccagtgtt tggatattca ggttcgtgaa actcaagctt tgatcttggc 1021 tcccacaaga gagttggctg tgcagatcca gaaggggctg cttgctctcg gtgactacat 1081 gaatgtccag tgccatgcct gcattggagg caccaatgtt ggcgaggaca tcaggaagct 1141 ggattacgga cagcatgttg tcgcgggcac tccagggcgt gtttttgata tgattcgtcg 1201 cagaagccta aggacacgtg ctatcaaaat gttggttttg gatgaagctg atgaaatgtt 1261 gaataaaggt ttcaaagagc agatttacga tgtatacagg tacctgcctc cagccacaca 1321 ggtggttctc atcagtgcca cgctgccaca cgagattctg gagatgacca acaagttcat 1381 gaccgaccca atccgcatct tggtgaaacg tgatgaattg actctggaag gcatcaagca 1441 atttttcgtg gcagtggaga gggaagagtg gaaatttgac actctgtgtg acctctacga 1501 cacactgacc atcactcagg cggtcatctt ctgcaacacc aaaagaaagg tggactggct 1561 gacggagaaa atgagggaag ccaacttcac tgtatcctca atgcatggag acatgcccca 1621 gaaagagcgg gagtccatca tgaaggagtt ccggtcgggc gccagccgag tgcttatttc 1681 tacagatgtc tgggccaggg ggttggatgt ccctcaggtg tccctcatca ttaactatga 1741 tctccctaat aacagagaat tgtacataca cagaattggg agatcaggtc gatacggccg 1801 gaagggtgtg gccattaact ttgtaaagaa tgacgacatc cgcatcctca gagatatcga 1861 gcagtactat tccactcaga ttgatgagat gccgatgaac gttgctgatc ttatcctcga 1921 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga 1981 tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa 2041 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta 2101 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa 2161 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg 2221 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc 2281 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc 2341 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg 2401 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg 2461 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg 2521 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc 2581 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa 2641 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt 2701 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca 2761 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca 2821 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca 2881 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 2941 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 3001 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 3061 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 3121 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 3181 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 3241 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat 3301 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 3361 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3421 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 3481 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 3541 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 3601 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 3661 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat 3721 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 3781 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 3841 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 3901 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc 3961 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 4021 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 4081 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 4141 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 4201 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 4261 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg 4321 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 4381 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 4441 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 4501 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 4561 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 4621 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 4681 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct 4741 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 4801 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg 4861 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 4921 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 4981 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 5041 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 5101 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 5161 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 5221 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 5281 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 5341 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 5401 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 5461 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg 5521 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 5581 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 5641 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 5701 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg 5761 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 5821 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 5881 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 5941 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 6001 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 6061 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 6121 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 6181 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 6241 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 6301 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 6361 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 6421 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 6481 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 6541 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 6601 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 6661 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 6721 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 6781 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 6841 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 6901 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 6961 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 7021 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat 7081 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg 7141 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag 7201 gcttgaccga caattgcatg aagaatctgc ttagg //