LOCUS V035490 7507 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035490 VERSION V035490 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..2997 /label="DHX58(NM_024119)" /note="DHX58(NM_024119)" /gene="DHX58" CDS 3004..3069 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3107..3331 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3377..3805 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3819..4148 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4215..5009 /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 5183..5304 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5353..5369) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5377..5393 /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(5401..5431) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5446..5467 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5755..6340) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6511..7371) /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(7372..7476) /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 accatggagc ttcggtccta ccaatgggag gtgatcatgc ctgccctgga gggcaagaat 1021 atcatcatct ggctgcccac gggtgccggg aagacccggg cggctgctta tgtggccaag 1081 cggcacctag agactgtgga tggagccaag gtggttgtat tggtcaacag ggtgcacctg 1141 gtgacccagc atggtgaaga gttcaggcgc atgctggatg gacgctggac cgtgacaacc 1201 ctgagtgggg acatgggacc acgtgctggc tttggccacc tggcccggtg ccatgacctg 1261 ctcatctgca cagcagagct tctgcagatg gcactgacca gccccgagga ggaggagcac 1321 gtggagctca ctgtcttctc cctgatcgtg gtggatgagt gccaccacac gcacaaggac 1381 accgtctaca acgtcatcat gagccagtac ctagaactta aactccagag ggcacagccg 1441 ctaccccagg tgctgggtct cacagcctcc ccaggcactg gcggggcctc caaactcgat 1501 ggggccatca accacgtcct gcagctctgt gccaacttgg acacgtggtg catcatgtca 1561 ccccagaact gctgccccca gctgcaggag cacagccaac agccttgcaa acagtacaac 1621 ctctgccaca ggcgcagcca ggatccgttt ggggacttgc tgaagaagct catggaccaa 1681 atccatgacc acctggagat gcctgagttg agccggaaat ttgggacgca aatgtatgag 1741 cagcaggtgg tgaagctgag tgaggctgcg gctttggctg ggcttcagga gcaacgggtg 1801 tatgcgcttc acctgaggcg ctacaatgac gcgctgctca tccatgacac cgtccgcgcc 1861 gtggatgcct tggctgcgct gcaggatttc tatcacaggg agcacgtcac taaaacccag 1921 atcctgtgtg ccgagcgccg gctgctggcc ctgttcgatg accgcaagaa tgagctggcc 1981 cacttggcaa ctcatggccc agagaatcca aaactggaga tgctggaaaa gatcctgcaa 2041 aggcagttca gtagctctaa cagccctcgg ggtatcatct tcacccgcac ccgccaaagc 2101 gcacactccc tcctgctctg gctccagcag cagcagggcc tgcagactgt ggacatccgg 2161 gcccagctac tgattggggc tgggaacagc agccagagca cccacatgac ccagagggac 2221 cagcaagaag tgatccagaa gttccaagat ggaaccctga accttctggt ggccacgagt 2281 gtggcggagg aggggctgga catcccacat tgcaatgtgg tggtgcgtta tgggctcttg 2341 accaatgaaa tctccatggt ccaggccagg ggccgtgccc gggccgatca gagtgtatac 2401 gcgtttgtag caactgaagg tagccgggag ctgaagcggg agctgatcaa cgaggcgctg 2461 gagacgctga tggagcaggc agtggctgct gtgcagaaaa tggaccaggc cgagtaccag 2521 gccaagatcc gggatctgca gcaggcagcc ttgaccaagc gggcggccca ggcagcccag 2581 cgggagaacc agcggcagca gttcccagtg gagcacgtgc agctactctg catcaactgc 2641 atggtggctg tgggccatgg cagcgacctg cggaaggtgg agggcaccca ccatgtcaat 2701 gtgaacccca acttctcgaa ctactataat gtctccaggg atcctgtggt catcaacaaa 2761 gtcttcaagg actggaagcc tgggggtgtc atcagctgca ggaactgtgg ggaggtctgg 2821 ggtctgcaga tgatctacaa gtcagtgaag ctgccagtgc tcaaagtccg cagcatgctg 2881 ctggagaccc ctcaggggcg gatccaggcc aaaaagtggt cccgcgtgcc cttctccgtg 2941 cctgactttg acttcctgca gcattgtgcc gagaacttgt cggacctctc cctggacctc 3001 gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 3061 gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt 3121 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 3181 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat 3241 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc 3301 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc 3361 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 3421 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 3481 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 3541 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 3601 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 3661 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 3721 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 3781 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 3841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3901 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3961 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 4021 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 4081 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 4141 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat 4201 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 4261 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 4321 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 4381 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 4441 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 4501 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 4561 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 4621 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 4681 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 4741 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 4801 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 4861 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 4921 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 4981 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca 5041 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 5101 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 5161 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag 5221 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 5281 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt 5341 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 5401 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 5461 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 5521 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 5581 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 5641 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 5701 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 5761 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 5821 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 5881 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 5941 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 6001 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 6061 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 6121 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 6181 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 6241 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt 6301 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 6361 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 6421 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 6481 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 6541 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 6601 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 6661 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 6721 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 6781 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 6841 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 6901 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 6961 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 7021 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 7081 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 7141 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 7201 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 7261 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 7321 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 7381 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 7441 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 7501 tgacgtc //