LOCUS V042348 6364 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042348 VERSION V042348 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 964..1854 /label="MEMO1(NM_015955)" /note="MEMO1(NM_015955)" /gene="MEMO1" CDS 1861..1926 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1964..2188 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2234..2662 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2676..3005 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3072..3866 /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 4040..4161 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4210..4226) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4234..4250 /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(4258..4288) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4303..4324 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4612..5197) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5368..6228) /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(6229..6333) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatgtcca accgagtggt ctgccgagaa gccagtcacg ccgggagctg gtacacagcc 1021 tcaggaccgc agctgaatgc acagctagaa ggttggcttt cacaagtaca gtctacaaaa 1081 agacctgcta gagccattat tgccccccat gcaggatata cgtactgtgg gtcttgtgct 1141 gcccatgctt ataaacaagt ggatccgtct attacccgga gaattttcat ccttgggcct 1201 tctcatcatg tgcccctctc tcgatgtgca ctttccagtg tggatatata taggacacct 1261 ctgtatgacc ttcgtattga ccaaaagatt tacggagaac tgtggaagac aggaatgttt 1321 gaacgcatgt ctctgcagac agatgaagat gaacacagta ttgaaatgca tttgccttat 1381 acagctaaag ccatggaaag ccataaggat gagtttacca ttattcctgt actggttgga 1441 gctctgagtg agtcaaaaga acaggaattc ggaaaactct tcagtaaata tctagcggat 1501 cctagtaatc tctttgtggt ttcttctgat ttctgccatt ggggtcaaag gttccgttac 1561 agttactatg atgaatccca gggggagatt tatagatcca ttgaacatct agataaaatg 1621 ggtatgagta ttatagaaca attagaccct gtatctttta gcaattactt gaagaaatac 1681 cataatacta tatgtggaag acatcccatt ggggtgttat taaatgctat cacagagctc 1741 cagaagaatg gaatgaatat gagtttttcg tttttgaatt atgcccagtc gagccagtgt 1801 agaaactggc aagacagttc agtgagttat gcagctggag cactcacggt ccacctcgag 1861 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat 1921 gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc 1981 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc 2041 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct 2101 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 2161 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct 2221 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 2281 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 2341 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 2401 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 2461 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 2521 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 2581 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 2641 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 2701 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 2761 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 2821 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 2881 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 2941 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 3001 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 3061 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 3121 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 3181 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 3241 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 3301 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 3361 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 3421 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 3481 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 3541 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 3601 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 3661 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 3721 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 3781 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 3841 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 3901 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 3961 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 4021 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa 4081 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 4141 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc 4201 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 4261 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 4321 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 4381 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 4441 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 4501 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 4561 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 4621 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 4681 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 4741 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 4801 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 4861 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 4921 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 4981 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 5041 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 5101 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg 5161 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 5221 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 5281 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 5341 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 5401 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 5461 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 5521 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 5581 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 5641 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 5701 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 5761 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 5821 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 5881 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 5941 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 6001 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 6061 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 6121 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 6181 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 6241 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 6301 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 6361 cgtc //