LOCUS V044109 7266 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044109 VERSION V044109 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" promoter 678..696 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 756..1946 /label="Dnaja1(NM_001164671)" /note="Dnaja1(NM_001164671)" /gene="Dnaja1" CDS 1953..2018 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2042..2630 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2696..2744 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2946..3374 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3388..3717 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3784..4578 /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 4754..4875 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4924..4940) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4948..4964 /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(4972..5002) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5017..5038 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5326..5914) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6085..6945) /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(6946..7050) /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 gctctctggc taactagaga acccactgct 661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta 721 aacttaagct tggtaccgag ctcgaattcg ccaccatggt gaaagaaacc acttactacg 781 atgttttggg ggtaaaaccc aatgccaccc aggaagaatt gaaaaaggca tatagaaaat 841 tggccttgaa gtaccaccct gataagaatc caaatgaagg ggaaaagttt aaacagattt 901 ctcaagctta tgaagttctt gctgattcca aaaaaaggga actatatgat aaaggagggg 961 agcaggcgat taaagagggc ggagcaggtg gtggttttgg ctcacccatg gatatctttg 1021 atatgttctt tggaggagga ggaagaatgc aaagagaaag gagaggtaaa aatgttgttc 1081 atcagctctc agtgacctta gaagacttat ataatggtgc aacaagaaaa ctggctctgc 1141 aaaagaatgt gatttgtgac aaatgtgaag gccgaggtgg taagaaagga gcagtagagt 1201 gctgtcccaa ctgccggggg acaggtatgc agataaggat tcatcagatt ggaccaggaa 1261 tggttcagca aattcagtca gtgtgcatgg agtgccaggg tcatggagaa cgcatcagtc 1321 caaaagacag atgtaaaagc tgcaatggaa gaaaaatagt tcgagagaag aaaattttag 1381 aagttcatat tgataaaggc atgaaagatg gtcagaagat aacattccac ggtgaaggag 1441 accaagaacc aggactggag ccaggagata ttatcattgt gttagatcag aaggaccatg 1501 ctgtttttac aaggcgagga gaagaccttt tcatgtgtat ggacatacag ctggttgaag 1561 cattgtgcgg cttccaaaag ccaatatcta ctcttgacaa ccgaaccata gtcatcacct 1621 ctcatccagg tcagattgtc aagcatgggg atataaaatg tgtgctaaat gaaggtatgc 1681 caatataccg tcggccatat gaaaagggac gtctaatcat tgagtttaag gtaaactttc 1741 ctgaaaatgg ctttctctct cctgataaac tctctttgct ggaaaaactc cttcctgaaa 1801 ggaaggaagt agaagagact gatgaaatgg atcaggtaga actggtggac tttgatccaa 1861 atcaggaaag acggcgtcat tataatggag aagcgtatga ggatgatgaa catcacccca 1921 gaggtggcgt tcagtgtcag acctctggat ccgactacaa agaccatgac ggtgattata 1981 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga 2041 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 2101 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 2161 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 2221 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 2281 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 2341 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 2401 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct 2461 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 2521 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 2581 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg 2641 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca 2701 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg 2761 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg 2821 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc 2881 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta 2941 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 3001 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 3061 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg 3121 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 3181 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 3241 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 3301 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 3361 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 3421 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 3481 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 3541 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 3601 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 3661 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 3721 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 3781 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 3841 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 3901 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 3961 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 4021 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 4081 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 4141 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 4201 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 4261 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 4321 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 4381 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 4441 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 4501 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 4561 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc 4621 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg 4681 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt 4741 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 4801 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 4861 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc 4921 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 4981 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 5041 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 5101 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 5161 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 5221 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 5281 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 5341 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 5401 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 5461 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 5521 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 5581 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 5641 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 5701 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 5761 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 5821 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 5881 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5941 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 6001 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 6061 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 6121 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 6181 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 6241 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 6301 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 6361 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 6421 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 6481 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 6541 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 6601 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 6661 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 6721 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 6781 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 6841 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 6901 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6961 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 7021 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 7081 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat 7141 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc 7201 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg 7261 cttagg //