LOCUS V040047 6541 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040047 VERSION V040047 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 928..1977 /label="FPR1(NM_001193306)" /note="FPR1(NM_001193306)" /gene="FPR1" CDS 1984..2007 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 2014..2103 /codon_start=1 /product="two Strep-Tag®II moieties connected by a linker for enhanced binding to Strep-Tactin®, an engineered form of streptavidin (Schmidt et al., 2013)" /transl_table=1 /translation="SA,WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK" /label="Twin-Strep-tag" polyA_signal 2141..2365 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2411..2839 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2853..3182 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3249..4043 /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 4217..4338 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4387..4403) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4411..4427 /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(4435..4465) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4480..4501 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4789..5374) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5545..6405) /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(6406..6510) /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 aagctgaatt cgccaccatg gagacaaatt cctctctccc cacgaacatc 961 tctggaggga cacctgctgt atctgctggc tatctcttcc tggatatcat cacttatctg 1021 gtatttgcag tcacctttgt cctcggggtc ctgggcaacg ggcttgtgat ctgggtggct 1081 ggattccgga tgacacacac agtcaccacc atcagttacc tgaacctggc cgtggctgac 1141 ttctgtttca cctccacttt gccattcttc atggtcagga aggccatggg aggacattgg 1201 cctttcggct ggttcctgtg caaattcgtc tttaccatag tggacatcaa cttgttcgga 1261 agtgtcttcc tgatcgccct cattgctctg gaccgctgtg tttgcgtcct gcatccagtc 1321 tggacccaga accaccgcac cgtgagcctg gccaagaagg tgatcattgg gccctgggtg 1381 atggctctgc tcctcacatt gccagttatc attcgtgtga ctacagtacc tggtaaaacg 1441 gggacagtag cctgcacttt taacttttcg ccctggacca acgaccctaa agagaggata 1501 aatgtggccg ttgccatgtt gacggtgaga ggcatcatcc ggttcatcat tggcttcagc 1561 gcacccatgt ccatcgttgc tgtcagttat gggcttattg ccaccaagat ccacaagcaa 1621 ggcttgatta agtccagtcg tcccttacgg gtcctctcct ttgtcgcagc agcctttttt 1681 ctctgctggt ccccatatca ggtggtggcc cttatagcca cagtcagaat ccgtgagtta 1741 ttgcaaggca tgtacaaaga aattggtatt gcagtggatg tgacaagtgc cctggccttc 1801 ttcaacagct gcctcaaccc catgctctat gtcttcatgg gccaggactt ccgggagagg 1861 ctgatccacg cccttcccgc cagtctggag agggccctga ccgaggactc aacccaaacc 1921 agtgacacag ctaccaattc tactttacct tctgcagagg tggagttaca ggcaaagctc 1981 gaggattaca aggatgacga cgataaggga tccagcgcct ggtcccaccc tcagtttgag 2041 aagggcggag gctctggcgg cggaagcgga ggatctgctt ggagccaccc ccagttcgaa 2101 aagtgatgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag 2161 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 2221 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 2281 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 2341 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg 2401 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 2461 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 2521 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 2581 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 2641 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 2701 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 2761 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 2821 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 2881 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 2941 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3001 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 3061 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 3121 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 3181 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat 3241 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 3301 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 3361 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 3421 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 3481 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 3541 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 3601 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 3661 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 3721 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca 3781 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 3841 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 3901 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 3961 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 4021 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 4081 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 4141 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 4201 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag 4261 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 4321 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta 4381 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 4441 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 4501 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 4561 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 4621 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 4681 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 4741 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 4801 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 4861 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 4921 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 4981 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 5041 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 5101 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 5161 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 5221 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 5281 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa 5341 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5401 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 5461 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5521 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5581 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 5641 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 5701 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 5761 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 5821 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 5881 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 5941 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 6001 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 6061 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 6121 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 6181 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 6241 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 6301 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 6361 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6421 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6481 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 6541 c //