LOCUS Exported 8658 bp ds-DNA circular SYN 30-OCT-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8658) AUTHORS MReijns TITLE Direct Submission JOURNAL Exported Oct 30, 2018 from SnapGene Viewer 4.2.4 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8658 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 579..920 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 987..1403 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" protein_bind 1429..1553 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 1578..1608 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 1662..2321 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" CDS 2663..2968 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" protein_bind complement(3009..3133) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 3151..3650 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" CDS 3668..4693 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" LTR 5051..5565 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 5727..5743 /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(5751..5781) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5796..5817 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(6105..6693) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6864..7724) /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(7725..7829) /gene="bla" /label=AmpR promoter ORIGIN 1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat 61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca 121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga 181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt 241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc 301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc 361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata 421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca 481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga 541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg 601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt 661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa 721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga 781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag 841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa 901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg 961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct 1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga 1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag 1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc 1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc 1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg 1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag 1381 ccctcactcc ttctctaggc gccggaatta gatctctcga ggttatcaac aagtttgtac 1441 aaaaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg 1501 cataaaaaac agactacata atactgtaaa acacaacata tccagtcact atggcggccg 1561 cattaggcac cccaggcttt acactttatg cttccggctc gtataatgtg tggattttga 1621 gttaggatcc gtcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg 1681 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt 1741 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga 1801 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga 1861 tgaatgctca tccggaattc cgtatggcaa tgaaagacgg tgagctggtg atatgggata 1921 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga 1981 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt 2041 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag 2101 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct 2161 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc 2221 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg 2281 aattacaaca gtactgcgat gagtggcagg gcggggcgta aagatctgga tccggcttac 2341 taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata agaatatata 2401 ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc gtattacagt 2461 gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa tatctccggt 2521 ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg ctggaaagcg 2581 gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg ctcttttgct 2641 gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa gagagagccg 2701 ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc gacggatggt 2761 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt 2821 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt 2881 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc 2941 cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca gccagtctgc 3001 aggtcgacca tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat 3061 gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt tcagctttct 3121 tgtacaaagt ggttgataac gaattctacc gggtagggga ggcgcttttc ccaaggcagt 3181 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc 3241 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct 3301 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt 3361 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca 3421 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct 3481 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc 3541 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa 3601 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcatcc 3661 cgccaccatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa 3721 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag 3781 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta 3841 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct 3901 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt 3961 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc 4021 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc 4081 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca 4141 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct 4201 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga 4261 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag 4321 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg 4381 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg 4441 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt 4501 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg 4561 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac 4621 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag 4681 ggcaaaggaa tagagtagat gccgaccgaa caagagctga tttcgagaac gcctcagcca 4741 gcaactcgcg cgagcctagc aaggcaaatg cgagagaacg gccttacgct tggtggcaca 4801 gttctcgtcc acagttcgct aagctcgctc ggctgggtcg cgggagggcc ggtcgcagtg 4861 attcaggccc ttctggattg tgttggtccc cagggcacga ttgtcatgcc cacgcactcg 4921 ggtgatctga ctgatcccgc agattggaga tcgccgcccg tgcctgccga ttgggtgcag 4981 atccgtcgac ctgcagccaa gcttatcgat aaaataaaag attttattta gtctccagaa 5041 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt 5101 ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca 5161 gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct 5221 cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc 5281 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa 5341 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga 5401 gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg 5461 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag 5521 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt 5581 gaagttggag aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt 5641 agccactgtt ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc 5701 agaaagagct ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga 5761 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 5821 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 5881 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 5941 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 6001 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 6061 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 6121 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 6181 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 6241 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 6301 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 6361 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 6421 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 6481 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 6541 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 6601 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 6661 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 6721 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 6781 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 6841 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 6901 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 6961 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 7021 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 7081 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 7141 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 7201 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 7261 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 7321 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 7381 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 7441 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 7501 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 7561 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 7621 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 7681 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 7741 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 7801 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 7861 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt 7921 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg 7981 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg 8041 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga 8101 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg 8161 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg 8221 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca 8281 gggttttccc agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga 8341 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc 8401 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc 8461 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat 8521 tagtccaatt tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat 8581 atcaccagct gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc 8641 tccagaaaaa ggggggaa //