LOCUS V046561 8935 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046561 VERSION V046561 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..4425 /label="PDZD8(NM_173791)" /note="PDZD8(NM_173791)" /gene="PDZD8" CDS 4432..4497 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 4535..4759 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4805..5233 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5247..5576 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5643..6437 /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 6611..6732 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6781..6797) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6805..6821 /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(6829..6859) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6874..6895 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7183..7768) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7939..8799) /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(8800..8904) /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 accatggggc tgctgctcat gatcctggcg tcggccgtgc tgggttcctt cctcacgctc 1021 ctcgcccagt tcttcctgct gtaccgcaga cagcccgagc cgccggcgga cgaggccgcc 1081 cgcgcgggcg agggcttccg ctacatcaag ccagtgccgg gcctgctcct aagggagtac 1141 ctttatggcg gcggccggga tgaggagccc tccggagcgg cccctgaggg cggcgcgacc 1201 cccaccgcgg cccccgagac ccccgccccg ccgacgcggg agacttgcta cttcctcaac 1261 gccaccatcc tattcctgtt ccgggagttg cgggacaccg cgctgacccg ccgctgggtc 1321 accaagaaga tcaaggtgga gttcgaggag ctgctgcaga ccaagacggc cgggcgcctg 1381 ctggaggggc tgagcctgcg ggacgtgttc ctgggcgaga cggtgccctt catcaagacc 1441 atccggctcg tgcggccagt cgtgccctcg gccaccgggg agcccgatgg ccctgaaggg 1501 gaggcgctgc ccgccgcctg ccccgaggag ctggccttcg aggcggaggt ggagtacaac 1561 gggggcttcc acctggccat cgacgtggac ctggtcttcg gcaagtccgc ctacttgttt 1621 gtcaagctgt cccgcgtggt gggaaggctg cgcttggtct ttacgcgcgt gcccttcacc 1681 cactggttct tctccttcgt ggaagacccg ctgatcgact tcgaggtgcg ctcccagttt 1741 gaagggcggc ccatgcccca gctcacctcc atcatcgtca accagctcaa gaagatcatc 1801 aagcgcaagc acaccctacc gaattacaag atcaggttta agccgttttt tccataccag 1861 accttgcaag gatttgaaga agatgaagag catatccata tacaacaatg ggcacttact 1921 gaaggccgtc ttaaagttac gttgttagaa tgtagcaggt tactcatttt tggatcctat 1981 gacagagagg caaatgttca ttgcacactt gagttaagca gtagtgtttg ggaagaaaaa 2041 cagaggagtt ctattaagac ggttgaatta ataaaaggaa atttacaaag tgttggactt 2101 acacttcgtc ttgtccagtc aactgatggg tatgctgggc acgtcatcat tgaaactgtg 2161 gctccaaact cgcctgctgc aattgcagat cttcagcggg gagatcgact tatcgccatt 2221 ggaggtgtga aaatcacatc aacactgcaa gtgttgaagc ttatcaagca ggctggtgac 2281 cgagtcctgg tgtactatga aaggcctgtt ggccagagta atcaaggtgc agtgctgcaa 2341 gataactttg gccagttgga agaaaacttt ttgtcaagct catgccaatc gggttatgaa 2401 gaggaagctg ccgggttgac agtagatact gaaagtagag agctggattc tgaatttgaa 2461 gacttggcaa gtgatgtcag agcacaaaat gagttcaaag atgaggcaca atcattaagt 2521 catagtccca aacgtgttcc aacaacactt tctattaaac cccttggagc tatatcacca 2581 gttttaaacc gtaaattagc tgtaggaagt cacccactac caccgaaaat tcagtccaaa 2641 gatggaaata aacctccacc cctaaaaact tctgagataa cagacccagc acaagtgtca 2701 aaaccaaccc aaggatctgc tttcaaacca cctgtgccac cacgaccaca agcgaaagtt 2761 cctttgcctt ccgccgatgc tccaaatcag gcagaaccag atgttctcgt tgaaaagcca 2821 gagaaggtgg tgccacctcc tcttgtagat aaatctgctg aaaagcaagc aaaaaatgtg 2881 gatgccatag acgatgcagc tgcacctaag caatttttag caaagcaaga agtggccaaa 2941 gatgtcactt cagaaacttc ctgccctact aaggacagtt cggacgaccg tcaaacatgg 3001 gaatcatcag aaattcttta tcgtaataag ctaggaaaat ggacaagaac cagagcatcc 3061 tgtttgtttg acatagaagc ctgtcacagg tacttaaaca ttgcattgtg gtgcagggat 3121 cctttcaagt tgggaggtct catctgtttg gggcatgtta gtttaaaact tgaagatgtg 3181 gctttaggat gcctagctac atcaaacacg gaataccttt ccaaattgag actggaagcc 3241 ccctcaccta aggctatagt cactagaacc gcactacgca atctgagtat gcaaaaggga 3301 ttcaatgaca aattttgcta tggtgacatt actattcact tcaaatattt gaaagaagga 3361 gaatcagacc accatgtagt tactaacgta gaaaaagaaa aagaacccca tttggttgaa 3421 gaagtttctg ttctccctaa agaggagcaa tttgttggac agatgggttt aacagaaaac 3481 aaacacagtt ttcaggatac tcagttccag aacccaacat ggtgtgacta ctgtaagaaa 3541 aaagtttgga ctaaagcagc ttcccagtgt atgttttgtg cttatgtttg ccataaaaaa 3601 tgtcaagaaa agtgtctagc tgagacttct gtttgtggag caactgatag gcgaatagac 3661 aggacactga aaaaccttag gctggaagga caggaaaccc tcttaggcct gcctcctcgt 3721 gttgatgctg aagctagcaa gtcagtcaat aaaacaacag gtttgacaag gcatattatc 3781 aatactagtt ctcgtttatt aaatttgcgt caagtctcta aaactcgcct ttctgaacca 3841 ggaaccgatc tcgtagaacc ttcaccaaaa cacacaccca acacgtcaga caacgaaggc 3901 agtgacacgg aggtctgtgg tccaaacagt ccttctaaac ggggaaacag cacaggaata 3961 aagttagtga gaaaagaggg tggtctggat gacagtgttt tcattgcagt taaagaaatt 4021 ggtcgtgatc tgtacagggg cttgcctaca gaggaaagga tccagaaact agagttcatg 4081 ttggataagc tacagaatga aattgatcag gagttggaac acaataattc ccttgttaga 4141 gaagaaaaag agacaactga tacaaggaaa aaatcacttc tttctgctgc cttagctaaa 4201 tcaggtgaaa ggctacaagc tctaacactt cttatgattc actacagagc aggcattgaa 4261 gatatagaaa ctttagaaag tctgtcttta gaccagcact ccaaaaaaat aagcaagtac 4321 acagatgata cagaagaaga ccttgataat gaaataagcc aactaataga ctctcagcca 4381 ttcagcagca tatcagatga cttatttggc ccatccgagt ctgtgctcga ggactacaaa 4441 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 4501 tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct 4561 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 4621 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 4681 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 4741 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat 4801 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 4861 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 4921 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 4981 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 5041 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 5101 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 5161 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 5221 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 5281 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 5341 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 5401 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 5461 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 5521 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 5581 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 5641 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 5701 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 5761 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 5821 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 5881 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 5941 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 6001 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 6061 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 6121 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 6181 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 6241 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 6301 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 6361 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 6421 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 6481 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 6541 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 6601 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 6661 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 6721 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg 6781 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6841 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6901 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6961 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 7021 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7081 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7141 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7201 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7261 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7321 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7381 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7441 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7501 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7561 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7621 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7681 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca 7741 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 7801 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 7861 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 7921 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 7981 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 8041 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 8101 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 8161 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 8221 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 8281 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 8341 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 8401 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 8461 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 8521 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 8581 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 8641 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 8701 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 8761 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 8821 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 8881 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //