LOCUS Exported 7149 bp ds-DNA circular SYN 17-MAY-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7149) 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..7149 /organism="synthetic DNA construct" /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" primer_bind 907..932 /label=JBPR62 CDS 913..1476 /codon_start=1 /label=Rac1 WT /translation="MQAIKCVVVGDGAVGKNCLLISYTTNAFPGEYIPTVFDNYSANVM VDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHH CPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQR GLKTVFDEAIRAVLCPPPVKKRKRK" primer_bind 937..987 /label=JBPR60 primer_bind complement(937..987) /label=JBPR61 primer_bind complement(1457..1483) /label=JBPR63 CDS 1501..2466 /codon_start=1 /label=BirA* /translation="MDKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLR DWGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKS GDACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLR KLGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQ GWITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLII GDKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK" primer_bind 1501..1521 /label=JBPR1 primer_bind 2425..2446 /label=JBPR3 primer_bind complement(2443..2466) /label=JBPR6 primer_bind complement(2449..2466) /label=JBPR2 CDS 2497..2523 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 2749..2973 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3019..3447 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3461..3790 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3641..3776 /label=SV40 ori /note="SV40 origin of replication" CDS 3857..4651 /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 4825..4946 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4995..5011) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5019..5035 /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(5043..5073) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5088..5109 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5397..5982) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6153..7013) /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(7014..7118) /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 gtatgcaggc catcaagtgt gtggtggtgg gagacggagc tgtaggtaaa 961 aattgcctac tgatcagtta cacaaccaat gcatttcctg gagaatatat ccctactgtc 1021 tttgacaatt attctgccaa tgttatggta gatggaaaac cggtgaatct gggcttatgg 1081 gatacagctg gacaagaaga ttatgacaga ttacgccccc tatcctatcc gcaaacagat 1141 gtgttcttaa tttgcttttc ccttgtgagt cctgcatcat ttgaaaatgt ccgtgcaaag 1201 tggtatcctg aggtgcggca ccactgtccc aacactccca tcatcctagt gggaactaaa 1261 cttgatctta gggatgataa agacacgatc gagaaactga aggagaagaa gctgactccc 1321 atcacctatc cgcagggtct agccatggct aaggagattg gtgctgtaaa atacctggag 1381 tgctcggcgc tcacacagcg aggcctcaag acagtgtttg acgaagcgat ccgagcagtc 1441 ctctgcccgc ctcccgtgaa gaagaggaag agaaaagaat tcggagccgg agctggtacc 1501 atggacaagg acaacaccgt gcccctgaag ctgatcgccc tgctggccaa cggcgagttc 1561 cactctggcg agcagctggg agagaccctg ggaatgagca gagccgccat caacaagcac 1621 atccagacac tgagagactg gggagtggac gtgttcaccg tgcctggcaa gggctacagc 1681 ctgcctgagc ctatccagct gctgaacgcc aagcagatcc tgggacagct ggatggcgga 1741 agcgtggccg tgctgcctgt gatcgactcc accaatcagt acctgctgga cagaatcgga 1801 gagctgaagt ccggcgacgc ctgcatcgcc gagtaccagc aggctggcag aggaggcaga 1861 ggacggaagt ggttcagccc attcggagcc aacctgtacc tgtccatgtt ctggagactg 1921 gagcagggac ctgctgctgc catcggactg agtctggtga tcggaatcgt gatggccgag 1981 gtgctgagaa agctgggagc cgacaaggtg agagtgaagt ggcctaatga cctgtacctc 2041 caggaccgca agctggctgg catcctggtg gagctgacag gcaagacagg cgatgccgct 2101 cagatcgtga tcggagccgg aatcaacatg gccatgagaa gagtggagga gagcgtggtg 2161 aaccagggct ggatcaccct gcaggaggct ggcatcaacc tggaccggaa caccctggcc 2221 gccatgctga tcagagagct gagagccgct ctggagctgt tcgagcagga gggactggct 2281 ccttacctga gcagatggga gaagctggac aacttcatca acagacctgt gaagctgatc 2341 atcggcgaca aggaaatctt cggcatctcc agaggaatcg acaagcaggg agctctgctg 2401 ctggagcagg acggaatcat caagccctgg atgggcggag aaatctccct gagaagcgca 2461 gagaagggat ccgtcgacgg agccggtgct ggcgcatacc cctacgacgt gcccgactac 2521 gcaggctccg cagttccagc ggtagaaatc acccacacct tctgggaaag gaacctgccc 2581 tctgtttctg gcctgctgaa gatcattgga ttttccacat cagtaactgc cctggggttt 2641 gtgctgtaca aatacaagct cctgccacgg tcttgacccg gggatatcca gcacagtggc 2701 ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta 2761 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca 2821 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc 2881 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata 2941 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg 3001 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 3061 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 3121 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 3181 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 3241 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 3301 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 3361 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 3421 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg 3481 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 3541 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 3601 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 3661 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 3721 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 3781 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg 3841 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg 3901 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc 3961 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg 4021 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt 4081 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg 4141 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat 4201 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca 4261 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca 4321 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa 4381 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa 4441 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc 4501 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga 4561 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc 4621 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac 4681 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 4741 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 4801 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa 4861 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 4921 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc 4981 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 5041 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 5101 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 5161 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 5221 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 5281 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 5341 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 5401 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 5461 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 5521 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 5581 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 5641 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 5701 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 5761 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 5821 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 5881 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt 5941 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 6001 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 6061 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 6121 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 6181 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 6241 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 6301 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 6361 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 6421 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 6481 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 6541 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 6601 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 6661 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 6721 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 6781 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 6841 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 6901 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 6961 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 7021 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 7081 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 7141 cctgacgtc //