LOCUS V023750 5762 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023750 VERSION V023750 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..1252 /label="PF4V1(NM_002620)" /note="PF4V1(NM_002620)" /gene="PF4V1" CDS 1259..1324 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1362..1586 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1632..2060 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2074..2403 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2470..3264 /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 3438..3559 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3608..3624) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3632..3648 /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(3656..3686) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3701..3722 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4010..4595) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4766..5626) /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(5627..5731) /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 atgagctccg cagccaggtc 961 ccgcctcacc cgcgccaccc gccaggagat gctgttcttg gcgttgctgc tcctgccagt 1021 tgtggtcgcc ttcgccagag ctgaagctga agaagatggg gacctgcagt gcctgtgtgt 1081 gaagaccacc tcccaggtcc gtcccaggca catcaccagc ctggaggtga tcaaggccgg 1141 accccactgc cccactgccc aactcatagc cacgctgaag aatgggagga aaatttgctt 1201 ggatctgcaa gccctgctgt acaagaaaat cattaaggaa catttggaga gtctcgagga 1261 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga 1321 caagtagtga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca 1381 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 1441 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 1501 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca 1561 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag 1621 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 1681 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 1741 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 1801 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 1861 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 1921 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 1981 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 2041 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 2101 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 2161 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 2221 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 2281 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 2341 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 2401 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga 2461 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 2521 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 2581 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 2641 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 2701 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 2761 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 2821 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 2881 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 2941 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 3001 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 3061 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 3121 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 3181 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 3241 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 3301 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 3361 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 3421 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 3481 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 3541 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 3601 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 3661 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 3721 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 3781 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 3841 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 3901 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 3961 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 4021 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 4081 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 4141 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 4201 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 4261 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 4321 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 4381 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 4441 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 4501 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca 4561 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 4621 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 4681 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 4741 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 4801 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 4861 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 4921 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 4981 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 5041 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 5101 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 5161 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 5221 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 5281 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 5341 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 5401 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 5461 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 5521 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 5581 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 5641 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 5701 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 5761 tc //