LOCUS Exported 7197 bp ds-DNA circular SYN 18-JAN-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 7197) AUTHORS MRC LMB TITLE Direct Submission JOURNAL Exported Wednesday, Jun 19, 2019 from SnapGene Viewer 4.3.8 https://www.snapgene.com FEATURES Location/Qualifiers source 1..7197 /organism="recombinant plasmid" /mol_type="other DNA" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 913..1518 /codon_start=1 /label=RalBGV /translation="MAANKSKGQSSLALHKVIMVGSVGVGKSALTLQFMYDEFVEDYEP TKADSYRKKVVLDGEEVQIDILDTAGQEDYAAIRDNYFRSGEGFLLVFSITEHESFTAT AEFREQILRVKAEEDKIPLLVVGNKSDLEERRQVPVEEARSKAEEWGVQYVETSAKTRA NVDKVFFDLMREIRTKKMSENKDKNGKKSSKNKKSFKER" CDS 1549..2514 /codon_start=1 /label=BirA* /translation="MDKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLR DWGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKS GDACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLR KLGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQ GWITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLII GDKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK" primer_bind 1549..1570 /label=JBPR73 (seq primer) primer_bind 2401..2418 /label=JBPR141 CDS 2545..2571 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 2797..3021 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3067..3495 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3509..3838 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3689..3824 /label=SV40 ori /note="SV40 origin of replication" CDS 3905..4699 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 4873..4994 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5043..5059) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5067..5083 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5091..5121) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5136..5157 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5445..6030) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6201..7061) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(7062..7166) /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 aagcttaccg gtatggctgc caacaagagt aagggccaga gctccttggc cctccacaag 961 gtgatcatgg ttggcagcgt aggcgttggc aagtcagccc tgacgcttca gttcatgtat 1021 gacgagtttg tagaagacta tgaacctacc aaagctgaca gttatagaaa gaaagtggtt 1081 cttgatgggg aagaagttca gatagatatt ctggacaccg ctgggcaaga ggactacgca 1141 gccattcgag ataactactt tcggagtggg gaagggtttc ttcttgtgtt ctcaatcaca 1201 gaacatgaat cctttacagc aactgccgaa ttcagggaac agattctccg tgtgaaggct 1261 gaagaagata aaattccact gctcgtcgtg ggaaacaagt ctgacctaga ggagcggagg 1321 caggtgcctg tggaggaggc caggagtaaa gccgaagagt ggggcgtgca gtacgtggag 1381 acgtcagcga agacccgggc caacgtggac aaggtgttct ttgacctaat gagagaaatc 1441 agaacaaaga agatgtcaga aaacaaagac aagaatggca agaaaagcag caagaacaag 1501 aaaagtttta aagaaagacc gcgggaattc ggagccggag ctggtaccat ggacaaggac 1561 aacaccgtgc ccctgaagct gatcgccctg ctggccaacg gcgagttcca ctctggcgag 1621 cagctgggag agaccctggg aatgagcaga gccgccatca acaagcacat ccagacactg 1681 agagactggg gagtggacgt gttcaccgtg cctggcaagg gctacagcct gcctgagcct 1741 atccagctgc tgaacgccaa gcagatcctg ggacagctgg atggcggaag cgtggccgtg 1801 ctgcctgtga tcgactccac caatcagtac ctgctggaca gaatcggaga gctgaagtcc 1861 ggcgacgcct gcatcgccga gtaccagcag gctggcagag gaggcagagg acggaagtgg 1921 ttcagcccat tcggagccaa cctgtacctg tccatgttct ggagactgga gcagggacct 1981 gctgctgcca tcggactgag tctggtgatc ggaatcgtga tggccgaggt gctgagaaag 2041 ctgggagccg acaaggtgag agtgaagtgg cctaatgacc tgtacctcca ggaccgcaag 2101 ctggctggca tcctggtgga gctgacaggc aagacaggcg atgccgctca gatcgtgatc 2161 ggagccggaa tcaacatggc catgagaaga gtggaggaga gcgtggtgaa ccagggctgg 2221 atcaccctgc aggaggctgg catcaacctg gaccggaaca ccctggccgc catgctgatc 2281 agagagctga gagccgctct ggagctgttc gagcaggagg gactggctcc ttacctgagc 2341 agatgggaga agctggacaa cttcatcaac agacctgtga agctgatcat cggcgacaag 2401 gaaatcttcg gcatctccag aggaatcgac aagcagggag ctctgctgct ggagcaggac 2461 ggaatcatca agccctggat gggcggagaa atctccctga gaagcgcaga gaagggatcc 2521 gtcgacggag ccggtgctgg cgcatacccc tacgacgtgc ccgactacgc aggctccgca 2581 gttccagcgg tagaaatcac ccacaccttc tgggaaagga acctgccctc tgtttctggc 2641 ctgctgaaga tcattggatt ttccacatca gtaactgccc tggggtttgt gctgtacaaa 2701 tacaagctcc tgccacggtc ttgacccggg gatatccagc acagtggcgg ccgctcgagt 2761 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat 2821 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc 2881 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg 2941 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg 3001 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt 3061 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 3121 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 3181 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 3241 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 3301 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 3361 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg 3421 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa 3481 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg 3541 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg 3601 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 3661 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca 3721 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc 3781 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa 3841 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt 3901 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct 3961 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct 4021 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga 4081 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc 4141 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg 4201 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc 4261 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca 4321 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga 4381 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc 4441 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga 4501 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca 4561 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg 4621 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct 4681 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc 4741 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 4801 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 4861 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 4921 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 4981 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 5041 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 5101 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 5161 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 5221 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 5281 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 5341 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 5401 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 5461 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 5521 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 5581 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 5641 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 5701 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 5761 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 5821 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 5881 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 5941 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag 6001 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 6061 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 6121 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 6181 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 6241 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 6301 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 6361 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 6421 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 6481 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 6541 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 6601 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 6661 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 6721 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 6781 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 6841 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 6901 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 6961 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 7021 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 7081 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 7141 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //