LOCUS V024517 6014 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024517 VERSION V024517 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..1504 /label="IFNA2(NM_000605)" /note="IFNA2(NM_000605)" /gene="IFNA2" CDS 1511..1576 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1614..1838 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1884..2312 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2326..2655 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2722..3516 /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 3690..3811 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3860..3876) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3884..3900 /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(3908..3938) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3953..3974 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4262..4847) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5018..5878) /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(5879..5983) /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 atggccttga cctttgcttt 961 actggtggcc ctcctggtgc tcagctgcaa gtcaagctgc tctgtgggct gtgatctgcc 1021 tcaaacccac agcctgggta gcaggaggac cttgatgctc ctggcacaga tgaggagaat 1081 ctctcttttc tcctgcttga aggacagaca tgactttgga tttccccagg aggagtttgg 1141 caaccagttc caaaaggctg aaaccatccc tgtcctccat gagatgatcc agcagatctt 1201 caatctcttc agcacaaagg actcatctgc tgcttgggat gagaccctcc tagacaaatt 1261 ctacactgaa ctctaccagc agctgaatga cctggaagcc tgtgtgatac agggggtggg 1321 ggtgacagag actcccctga tgaaggagga ctccattctg gctgtgagga aatacttcca 1381 aagaatcact ctctatctga aagagaagaa atacagccct tgtgcctggg aggttgtcag 1441 agcagaaatc atgagatctt tttctttgtc aacaaacttg caagaaagtt taagaagtaa 1501 ggaactcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa 1561 ggatgacgat gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc 1621 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 1681 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 1741 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 1801 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag 1861 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 1921 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 1981 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 2041 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 2101 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 2161 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 2221 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 2281 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg 2341 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 2401 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 2461 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 2521 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 2581 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 2641 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag 2701 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 2761 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 2821 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 2881 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 2941 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 3001 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 3061 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 3121 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 3181 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 3241 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 3301 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 3361 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 3421 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 3481 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga 3541 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 3601 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 3661 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 3721 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 3781 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 3841 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 3901 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 3961 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 4021 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 4081 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 4141 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 4201 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 4261 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 4321 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 4381 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 4441 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 4501 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 4561 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 4621 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 4681 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 4741 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt 4801 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 4861 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 4921 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 4981 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 5041 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 5101 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 5161 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 5221 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 5281 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 5341 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 5401 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 5461 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 5521 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 5581 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 5641 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 5701 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 5761 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 5821 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 5881 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 5941 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 6001 tgccacctga cgtc //