LOCUS V040043 6937 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040043 VERSION V040043 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..2373 /label="C3AR1(NM_001326475)" /note="C3AR1(NM_001326475)" /gene="C3AR1" CDS 2380..2403 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 2410..2499 /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 2537..2761 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2807..3235 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3249..3578 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3645..4439 /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 4613..4734 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4783..4799) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4807..4823 /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(4831..4861) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4876..4897 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5185..5770) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5941..6801) /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(6802..6906) /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 gcgtctttct ctgctgagac caattcaact 961 gacctactct cacagccatg gaatgagccc ccagtaattc tctccatggt cattctcagc 1021 cttacttttt tactgggatt gccaggcaat gggctggtgc tgtgggtggc tggcctgaag 1081 atgcagcgga cagtgaacac aatttggttc ctccacctca ccttggcgga cctcctctgc 1141 tgcctctcct tgcccttctc gctggctcac ttggctctcc agggacagtg gccctacggc 1201 aggttcctat gcaagctcat cccctccatc attgtcctca acatgtttgc cagtgtcttc 1261 ctgcttactg ccattagcct ggatcgctgt cttgtggtat tcaagccaat ctggtgtcag 1321 aatcatcgca atgtagggat ggcctgctct atctgtggat gtatctgggt ggtggctttt 1381 gtgatgtgca ttcctgtgtt cgtgtaccgg gaaatcttca ctacagacaa ccataataga 1441 tgtggctaca aatttggtct ctccagctca ttagattatc cagactttta tggagatcca 1501 ctagaaaaca ggtctcttga aaacattgtt cagccgcctg gagaaatgaa tgataggtta 1561 gatccttcct ctttccaaac aaatgatcat ccttggacag tccccactgt cttccaacct 1621 caaacatttc aaagaccttc tgcagattca ctccctaggg gttctgctag gttaacaagt 1681 caaaatctgt attctaatgt atttaaacct gctgatgtgg tctcacctaa aatccccagt 1741 gggtttccta ttgaagatca cgaaaccagc ccactggata actctgatgc ttttctctct 1801 actcatttaa agctgttccc tagcgcttct agcaattcct tctacgagtc tgagctacca 1861 caaggtttcc aggattatta caatttaggc caattcacag atgacgatca agtgccaaca 1921 cccctcgtgg caataacgat cactaggcta gtggtgggtt tcctgctgcc ctctgttatc 1981 atgatagcct gttacagctt cattgtcttc cgaatgcaaa ggggccgctt cgccaagtct 2041 cagagcaaaa cctttcgagt ggccgtggtg gtggtggctg tctttcttgt ctgctggact 2101 ccataccaca tttttggagt cctgtcattg cttactgacc cagaaactcc cttggggaaa 2161 actctgatgt cctgggatca tgtatgcatt gctctagcat ctgccaatag ttgctttaat 2221 cccttccttt atgccctctt ggggaaagat tttaggaaga aagcaaggca gtccattcag 2281 ggaattctgg aggcagcctt cagtgaggag ctcacacgtt ccacccactg tccctcaaac 2341 aatgtcattt cagaaagaaa tagtacaact gtgctcgagg attacaagga tgacgacgat 2401 aagggatcca gcgcctggtc ccaccctcag tttgagaagg gcggaggctc tggcggcgga 2461 agcggaggat ctgcttggag ccacccccag ttcgaaaagt gatgagggcc cgtttaaacc 2521 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 2581 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 2641 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 2701 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 2761 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 2821 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 2881 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 2941 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 3001 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 3061 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 3121 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 3181 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 3241 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 3301 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 3361 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3421 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 3481 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 3541 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 3601 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 3661 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 3721 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 3781 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 3841 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 3901 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 3961 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 4021 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 4081 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 4141 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 4201 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 4261 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 4321 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 4381 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 4441 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 4501 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 4561 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 4621 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 4681 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 4741 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 4801 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 4861 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 4921 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 4981 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 5041 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 5101 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 5161 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 5221 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 5281 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 5341 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 5401 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 5461 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 5521 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 5581 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 5641 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 5701 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5761 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5821 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5881 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 5941 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 6001 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 6061 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 6121 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 6181 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 6241 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 6301 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 6361 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 6421 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 6481 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6541 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 6601 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6661 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6721 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6781 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 6841 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 6901 tccgcgcaca tttccccgaa aagtgccacc tgacgtc //