LOCUS V023586 7076 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023586 VERSION V023586 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 941..2566 /label="ZNF280A(NM_080740)" /note="ZNF280A(NM_080740)" /gene="ZNF280A" CDS 2573..2638 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2676..2900 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2946..3374 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3388..3717 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3784..4578 /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 4752..4873 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4922..4938) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4946..4962 /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(4970..5000) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5015..5036 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5324..5909) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6080..6940) /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(6941..7045) /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 atccgccacc atgggagata tctttttgtg 961 taagaaagtg gaatcaccaa agaagaattt gagagaatcc aaacaaaggg aggaggatga 1021 tgaagatcca gatctgatct atgttggggt ggagcatgta catagagatg ctgaagttct 1081 ctttgtcggg atgatttcaa attcaaaacc agtcgtttca aacattttga acagagtcac 1141 cccaggctca aattcaagaa gaaagaaagg ccacttccgt caatatcctg ctcacgtgtc 1201 gcagcctgca aatcatgtga cctctatggc aaaagccatc atgccggttt ctctgtctga 1261 ggggcgatcg acagatagtc ctgtcactat gaagtcttca tctgaacctg gttataaaat 1321 gagctcacca caagttgttt ctcccagttc ctcagactcg ctccccccag ggactcagtg 1381 tctagttgga gctatggtct ctggaggagg cagaaatgag agttctcctg attcaaagcg 1441 actttccact tcagatataa acagcagaga ttccaaaagg gttaaactca gggatggaat 1501 cccaggggta ccttctttag ctgtggtccc ttcagatatg tcttctacaa taagcacaaa 1561 tacaccctca caggggatct gcaactcatc aaaccatgtt cagaatggag taacatttcc 1621 ttggcctgat gctaatggaa aggcacattt caatcttaca gatccagaga gagcaagtga 1681 gtctgccctg gcaatgacag acatttcaag tctagcaagt caaaacaaga cctttgatcc 1741 caagaaagaa aatcccatcg tgttacttag cgacttttac tatggacagc ataaaggaga 1801 tgggcagccg gaacagaaga ctcacaccac ctttaaatgc ctcagctgcg tgaaagttct 1861 aaaaaatatt aagtttatga atcacatgaa gcatcatttg gaatttgaga agcagaggaa 1921 cgacagctgg gaagaccaca ccacctgcca gcactgccac cggcagtttc ccactccctt 1981 ccagctacag tgtcacattg atagtgtaca catcgccatg gggccctctg ctgtctgtaa 2041 aatctgtgaa ttgtcatttg aaacagatca ggtcctctta caacacatga aggaccatca 2101 taagcctggc gaaatgcctt atgtgtgcca ggtttgccat tacagatcgt cggtctttgc 2161 tgatgtggaa acacatttta gaacgtgcca tgaaaacaca aagaatttgc tttgtctgtt 2221 ttgtctcaaa cttttcaaaa ctgcaatacc atacatgaat cattgttgga ggcacagcag 2281 aaggagggtc cttcagtgtt ccaagtgccg gctacagttt ttgacgttga aggaggaaat 2341 agagcacaaa accaaggacc atcaaacatt taaaaagccg gagcaactgc aagggttgcc 2401 tagtgaaaca aaagttatta ttcaaacttc agttcagcca ggatcaagtg gtatggcttc 2461 cgttattgtt agcaacactg accctcagtc ttctcctgta aaaactaaaa agaagacggc 2521 tatgaacact agagattcca gactcccttg cagcaaggat tctagcctcg aggactacaa 2581 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 2641 gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 2701 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 2761 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 2821 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 2881 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta 2941 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 3001 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 3061 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 3121 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 3181 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 3241 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 3301 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 3361 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 3421 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 3481 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 3541 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 3601 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 3661 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 3721 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 3781 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 3841 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 3901 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 3961 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 4021 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 4081 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 4141 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 4201 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 4261 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 4321 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 4381 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 4441 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 4501 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 4561 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 4621 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 4681 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 4741 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 4801 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 4861 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 4921 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 4981 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 5041 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 5101 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 5161 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 5221 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 5281 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 5341 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 5401 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 5461 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 5521 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 5581 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 5641 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 5701 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 5761 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 5821 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc 5881 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 5941 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 6001 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 6061 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 6121 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 6181 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 6241 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 6301 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 6361 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 6421 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 6481 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 6541 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 6601 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 6661 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 6721 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 6781 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 6841 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 6901 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 6961 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 7021 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc //