LOCUS V044047 7333 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044047 VERSION V044047 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..2823 /label="PRC1(NM_003981)" /note="PRC1(NM_003981)" /gene="PRC1" CDS 2830..2895 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2933..3157 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3203..3631 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3645..3974 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4041..4835 /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 5009..5130 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5179..5195) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5203..5219 /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(5227..5257) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5272..5293 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5581..6166) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6337..7197) /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(7198..7302) /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 accatgagga gaagtgaggt gctggcggag gagtccatag tatgtctgca gaaagcccta 1021 aatcaccttc gggaaatatg ggagctaatt gggattccag aggaccagcg gttacaaaga 1081 actgaggtgg taaagaagca tatcaaggaa ctcctggata tgatgattgc tgaagaggaa 1141 agcctgaagg aaagactcat caaaagcata tccgtctgtc agaaagagct gaacactctg 1201 tgcagcgagt tacatgttga gccatttcag gaagaaggag agacgaccat cttgcaacta 1261 gaaaaagatt tgcgcaccca agtggaattg atgcgaaaac agaaaaagga gagaaaacag 1321 gaactgaagc tacttcaaga gcaagatcaa gaactgtgcg aaattctttg tatgccccac 1381 tatgatattg acagtgcctc agtgcccagc ttagaagagc tgaaccagtt caggcaacat 1441 gtgacaactt tgagggaaac aaaggcttct aggcgtgagg agtttgtcag tataaagaga 1501 cagatcatac tgtgtatgga agcattagac cacaccccag acacaagctt tgaaagagat 1561 gtggtgtgtg aagacgaaga tgccttttgt ttgtctttgg agaatattgc aacactacaa 1621 aagttgctac ggcagctgga aatgcagaaa tcacaaaatg aagcagtgtg tgaggggctg 1681 cgtactcaaa tccgagagct ctgggacagg ttgcaaatac ctgaagaaga aagagaagct 1741 gtggccacca ttatgtctgg gtcaaaggcc aaggtccgga aagcgctgca attagaagtg 1801 gatcggttgg aagaactgaa aatgcaaaac atgaagaaag tgattgaggc aattcgagtg 1861 gagctggttc agtactggga ccagtgcttt tatagccagg agcagagaca agcttttgcc 1921 cctttctgtg ctgaggacta cacagaaagt ctgctccagc tccacgatgc tgagattgtg 1981 cggttaaaaa actactatga agttcacaag gaactctttg aaggtgtcca gaagtgggaa 2041 gaaacctgga ggcttttctt agagtttgag agaaaagctt cagatccaaa tcgatttaca 2101 aaccgaggag gaaatcttct aaaagaagaa aaacaacgag ccaagctcca gaaaatgctg 2161 cccaagctgg aagaagagtt gaaggcacga attgaattgt gggaacagga acattcaaag 2221 gcatttatgg tgaatgggca gaaattcatg gagtatgtgg cagaacaatg ggagatgcat 2281 cgattggaga aagagagagc caagcaggaa agacaactga agaacaaaaa acagacagag 2341 acagagatgc tgtatggcag cgctcctcga acacctagca agcggcgagg actggctccc 2401 aatacaccgg gcaaagcacg taagctgaac actaccacca tgtccaatgc tacggccaat 2461 agtagcattc ggcctatctt tggagggaca gtctaccact cccccgtgtc tcgacttcct 2521 ccttctggca gcaagccagt cgctgcttcc acctgttcag ggaagaaaac accccgtact 2581 ggcaggcatg gagccaacaa ggagaacctg gagctcaacg gcagcatcct gagtggtggg 2641 taccctggct cggcccccct ccagcgcaac ttcagcatta attctgttgc cagcacctat 2701 tctgagtttg cgaaggatcc gtccctctct gacagttcca ctgttgggct tcagcgagaa 2761 ctttcaaagg cttccaaatc tgatgctact tctggaatcc tcaattcaac caacatccag 2821 tccctcgagg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag 2881 gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc gactgtgcct 2941 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt 3001 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg 3061 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac 3121 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc 3181 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 3241 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 3301 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 3361 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 3421 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 3481 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 3541 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat 3601 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt 3661 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3721 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3781 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 3841 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 3901 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 3961 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga 4021 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg 4081 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 4141 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt 4201 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg 4261 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 4321 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 4381 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 4441 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 4501 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc 4561 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc 4621 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 4681 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 4741 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 4801 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac 4861 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga 4921 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga 4981 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa 5041 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 5101 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta 5161 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 5221 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 5281 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 5341 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 5401 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 5461 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 5521 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 5581 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 5641 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 5701 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 5761 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 5821 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 5881 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 5941 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6001 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 6061 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt 6121 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 6181 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 6241 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 6301 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 6361 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 6421 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 6481 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 6541 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 6601 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 6661 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 6721 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 6781 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 6841 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 6901 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 6961 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 7021 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 7081 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 7141 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 7201 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 7261 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 7321 gccacctgac gtc //