LOCUS V022837 5834 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022837 VERSION V022837 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..1324 /label="CD59(NM_203330)" /note="CD59(NM_203330)" /gene="CD59" CDS 1331..1396 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1434..1658 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1704..2132 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2146..2475 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2542..3336 /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 3510..3631 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3680..3696) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3704..3720 /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(3728..3758) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3773..3794 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4082..4667) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4838..5698) /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(5699..5803) /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 atgggaatcc aaggagggtc 961 tgtcctgttc gggctgctgc tcgtcctggc tgtcttctgc cattcaggtc atagcctgca 1021 gtgctacaac tgtcctaacc caactgctga ctgcaaaaca gccgtcaatt gttcatctga 1081 ttttgatgcg tgtctcatta ccaaagctgg gttacaagtg tataacaagt gttggaagtt 1141 tgagcattgc aatttcaacg acgtcacaac ccgcttgagg gaaaatgagc taacgtacta 1201 ctgctgcaag aaggacctgt gtaactttaa cgaacagctt gaaaatggtg ggacatcctt 1261 atcagagaaa acagttcttc tgctggtgac tccatttctg gcagcagcct ggagccttca 1321 tcccctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa 1381 ggatgacgat gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc 1441 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 1501 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 1561 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 1621 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag 1681 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 1741 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 1801 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 1861 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 1921 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 1981 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 2041 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 2101 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg 2161 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 2221 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 2281 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 2341 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 2401 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 2461 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag 2521 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 2581 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 2641 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 2701 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 2761 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 2821 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 2881 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 2941 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 3001 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 3061 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 3121 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 3181 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 3241 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 3301 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga 3361 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 3421 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 3481 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 3541 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 3601 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 3661 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 3721 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 3781 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 3841 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 3901 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 3961 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 4021 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 4081 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 4141 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 4201 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 4261 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 4321 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 4381 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 4441 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 4501 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 4561 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt 4621 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 4681 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 4741 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 4801 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 4861 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 4921 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 4981 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 5041 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 5101 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 5161 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 5221 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 5281 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 5341 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 5401 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 5461 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 5521 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 5581 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 5641 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 5701 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 5761 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 5821 tgccacctga cgtc //