LOCUS Exported 6925 bp ds-DNA circular SYN 16-JUN-2015 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS MEM149C SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6925) AUTHORS Leonard Lab TITLE Direct Submission JOURNAL Exported Jun 16, 2015 from SnapGene 2.7.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6925 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 110..489 /note="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 490..693 /note="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" promoter 738..756 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" CDS 785..868 /codon_start=1 /label=signal peptide coding region /note="signal peptide coding region" /translation="MVCFRLFPVPGSGLVLVCLVLGAVRSYA" misc_feature 869..883 /note="GNSTM glycosylation tag" misc_feature 884..892 /note="GSG spacer between glyc tag and flag" CDS 893..916 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /note="FLAG" /translation="DYKDDDDK" misc_feature 917..946 /note="10gs spacer" misc_feature 947..2092 /label=Lamp2b protein coding region(1) /note="Lamp2b protein coding region" CDS 2102..2128 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /note="HA" /translation="YPYDVPDYA" polyA_signal 2231..2455 /note="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" rep_origin 2501..2929 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2943..3272 /note="SV40 promoter" /note="SV40 enhancer and early promoter" rep_origin 3123..3258 /note="SV40 ori" /note="SV40 origin of replication" CDS 3321..4346 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /note="HygR" /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 4476..4597 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" primer_bind complement(4646..4662) /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind 4670..4686 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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(4694..4724) /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 4739..4760 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5048..5633) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5804..6664) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(6665..6769) /gene="bla" /note="AmpR promoter" ORIGIN 1 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg 61 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 361 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 421 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 481 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 541 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 601 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 661 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct 721 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagtgccgc 781 cgccatggtg tgcttccgcc tcttcccggt tccgggctca gggctcgttc tggtctgcct 841 agtcctggga gctgtgcggt cttatgcagg taactcgact atgggcagtg gagactataa 901 agatgacgat gacaagggca gtggatctgg atccggtggc tcgagtttgg aacttaattt 961 gacagattca gaaaatgcca cttgccttta tgcaaaatgg cagatgaatt tcacagttcg 1021 ctatgaaact acaaataaaa cttataaaac tgtaaccatt tcagaccatg gcactgtgac 1081 atataatgga agcatttgtg gggatgatca gaatggtccc aaaatagcag tgcagttcgg 1141 acctggcttt tcctggattg cgaattttac caaggcagca tctacttatt caattgacag 1201 cgtctcattt tcctacaaca ctggtgataa cacaacattt cctgatgctg aagataaagg 1261 aattcttact gttgatgaac ttttggccat cagaattcca ttgaatgacc tttttagatg 1321 caatagttta tcaactttgg aaaagaatga tgttgtccaa cactactggg atgttcttgt 1381 acaagctttt gtccaaaatg gcacagtgag cacaaatgag ttcctgtgtg ataaagacaa 1441 aacttcaaca gtggcaccca ccatacacac cactgtgcca tctcctacta caacacctac 1501 tccaaaggaa aaaccagaag ctggaaccta ttcagttaat aatggcaatg atacttgcct 1561 gctggctacc atggggctgc agctgaacat cactcaggat aaggttgctt cagttattaa 1621 catcaacccc aatacaactc actccacagg cagctgccgt tctcacactg ctctacttag 1681 actcaatagc agcactatta agtatctaga ctttgtcttt gctgtgaaaa atgaaaaccg 1741 attttatctg aaggaagtga acatcagcat gtatttggtt aatggctccg ttttcagcat 1801 tgcaaataac aatctcagct actgggatgc ccccctggga agttcttata tgtgcaacaa 1861 agagcagact gtttcagtgt ctggagcatt tcagataaat acctttgatc taagggttca 1921 gcctttcaat gtgacacaag gaaagtattc tacagcccaa gagtgttcgc tggatgatga 1981 caccattcta atcccaatta tagttggtgc tggtctttca ggcttgatta tcgttatagt 2041 gattgcttac gtaattggca gaagaaaaag ttatgctgga tatcagactc tgggaagcgg 2101 atacccatac gatgtgccag attacgctta aggatccact agtccagtgt ggtggaattc 2161 tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga 2221 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 2281 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 2341 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag 2401 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct 2461 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca 2521 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 2581 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 2641 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 2701 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 2761 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 2821 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 2881 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga 2941 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3001 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 3061 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 3121 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 3181 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag 3241 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt 3301 tcggatctga tcagcacgtg atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt 3361 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat 3421 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg 3481 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga 3541 ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc 3601 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc 3661 cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg 3721 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga 3781 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg 3841 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc 3901 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg 3961 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct 4021 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc 4081 cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac 4141 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg 4201 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg 4261 cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca 4321 gcactcgtcc gagggcaaag gaatagcacg tgctacgaga tttcgattcc accgccgcct 4381 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc 4441 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg 4501 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 4561 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct 4621 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 4681 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 4741 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 4801 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 4861 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 4921 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 4981 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 5041 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 5101 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 5161 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 5221 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 5281 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 5341 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 5401 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 5461 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 5521 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 5581 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 5641 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 5701 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 5761 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 5821 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 5881 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 5941 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 6001 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 6061 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 6121 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 6181 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 6241 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 6301 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 6361 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 6421 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 6481 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 6541 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 6601 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 6661 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 6721 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 6781 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct 6841 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt 6901 ggaggtcgct gagtagtgcg cgagc //