LOCUS V041750 7945 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041750 VERSION V041750 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..3435 /label="DDX20(NM_007204)" /note="DDX20(NM_007204)" /gene="DDX20" CDS 3442..3507 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3545..3769 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3815..4243 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4257..4586 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4653..5447 /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 5621..5742 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5791..5807) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5815..5831 /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(5839..5869) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5884..5905 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6193..6778) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6949..7809) /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(7810..7914) /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 accatggcgg cggcatttga agcctcggga gccttagcag cagtggcgac tgctatgccg 1021 gctgagcatg tggccgtgca ggtcccggcc ccagagccaa cacccgggcc tgtgaggatc 1081 ctgcggaccg ctcaggatct cagcagcccg cggacccgca cgggggatgt gctgttggcg 1141 gagccggccg acttcgagtc actgctgctt tcgcggccgg tgctggaggg gctgcgggcg 1201 gccggcttcg agaggccctc gccggtgcag ctcaaggcca tcccgttggg gcgctgcggg 1261 ctcgatttaa ttgttcaagc taaatctggc accgggaaaa cctgtgtgtt ctccaccata 1321 gctttggact ctcttgttct tgaaaactta agtacccaga ttttgatctt ggctcctaca 1381 agagaaattg ctgtacagat acattctgtt attacagcca ttggaataaa aatggaaggc 1441 ttagagtgtc atgtctttat tggagggacc ccattatcac aagacaaaac cagacttaaa 1501 aagtgtcata ttgctgttgg atctcctggc agaattaagc aactcataga acttgactac 1561 ttgaacccag gcagtatacg cctctttatt cttgatgaag cagataagct tttagaagaa 1621 ggcagcttcc aggagcaaat aaattggatt tattcttcct tgcctgccag taaacagatg 1681 ctggcagtat cagctactta tcccgaattt ttggctaatg ctttgacaaa gtacatgaga 1741 gatcccactt ttgtaagact gaattccagt gatccaagtc tcataggttt gaagcagtat 1801 tacaaagttg tcaattcata ccctttggca cataaggttt ttgaggaaaa gactcagcat 1861 ttacaggaac tgttcagcag aattccattt aatcaagctt tagtcttttc taatttgcac 1921 agcagagcac aacatttggc tgatatcctt tcttctaaag gctttcctgc tgagtgcatt 1981 tcaggcaata tgaatcagaa tcagcgtctt gatgctatgg ctaaactgaa gcactttcat 2041 tgcagagtcc tcatttccac agatttgact tctcgtggga ttgatgctga gaaggtgaat 2101 ctggttgtaa atctggatgt accattggat tgggagacat acatgcatcg gattgggaga 2161 gctggccgtt ttggtacatt ggggctgaca gtgacctact gttgccgggg agaggaagaa 2221 aatatgatga tgagaattgc ccagaaatgt aatatcaacc ttctcccttt accagatccc 2281 attccttctg gtctgatgga agaatgtgtg gattgggatg tggaagttaa agctgctgtg 2341 catacatatg gtatagcaag tgtacctaac caacccttaa aaaagcaaat tcagaaaata 2401 gagagaaccc ttcaaattca gaaagctcat ggtgaccaca tggcttcctc tagaaataat 2461 tctgtatctg gactatcagt caaatcaaaa aataatacca aacaaaagct tcctgtgaaa 2521 agccactcag aatgtggaat catagaaaaa gcaacgtcac caaaagaact gggctgtgac 2581 aggcaatccg aagagcaaat gaagaattct gttcagactc ccgttgaaaa ctccaccaac 2641 agtcagcacc aggtcaaaga agctttacct gtgtcactcc cccagattcc ttgtctgtct 2701 tcctttaaaa tccatcagcc atacacgttg acttttgctg aattggtaga ggattatgaa 2761 cattatatta aagaggggtt agagaaacct gtggaaatca tcaggcacta cacaggccct 2821 ggggatcaga ctgtgaatcc tcaaaatggt tttgtgagaa ataaagttat tgaacagaga 2881 gtccctgtgt tggcaagtag tagccaatct ggagactctg agagtgacag tgattcttac 2941 agctcaagaa cctcttccca gagcaaagga aataagtcat acttggaagg ctcttctgat 3001 aatcagctga aagactctga atctacgcct gtggatgatc gtatttcttt ggaacaacca 3061 ccaaatggaa gtgacacccc caatccagag aaatatcaag aatcacctgg aatccagatg 3121 aagacaagac ttaaagaggg ggctagccag agagctaagc agagccggag aaacctaccc 3181 aggcggtctt ccttcagatt gcagactgaa gcccaggaag atgattggta tgactgtcat 3241 agggaaatac gtctgagttt ttctgatacc tatcaggatt atgaggagta ctggagagct 3301 tactacaggg catggcaaga atattatgct gccgcttctc attcatatta ttggaatgct 3361 cagagacatc caagttggat ggcagcttat cacatgaata ccatttatct acaagaaatg 3421 atgcatagta accagctcga ggactacaaa gaccatgacg gtgattataa agatcatgac 3481 atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg ctgatcagcc 3541 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 3601 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 3661 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 3721 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 3781 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 3841 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 3901 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 3961 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 4021 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 4081 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 4141 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 4201 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 4261 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 4321 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 4381 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 4441 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 4501 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 4561 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat 4621 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 4681 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 4741 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 4801 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc 4861 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 4921 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 4981 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 5041 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 5101 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 5161 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg 5221 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 5281 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 5341 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 5401 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 5461 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc 5521 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 5581 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 5641 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 5701 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 5761 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 5821 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 5881 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 5941 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 6001 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 6061 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 6121 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 6181 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 6241 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 6301 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 6361 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 6421 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 6481 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 6541 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 6601 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 6661 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 6721 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 6781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 6841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 6901 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 6961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7021 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 7081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 7141 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 7201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 7261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 7321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 7381 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 7441 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 7501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 7561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 7621 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 7681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 7741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 7801 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 7861 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 7921 tccccgaaaa gtgccacctg acgtc //