LOCUS Exported 7977 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 7977) 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..7977 /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 3671..4270 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" LTR 4370..4884 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 5046..5062 /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(5070..5100) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5115..5136 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5424..6012) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6183..7043) /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(7044..7148) /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 acctgcagcc 3661 caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc 3721 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt 3781 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt 3841 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac 3901 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga 3961 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg 4021 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa 4081 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc 4141 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac 4201 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc 4261 cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca 4321 tgcatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc 4381 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca 4441 taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc 4501 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc 4561 cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc 4621 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct 4681 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc 4741 gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg 4801 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac 4861 tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct 4921 gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca 4981 tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg 5041 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 5101 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 5161 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 5221 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 5281 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 5341 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 5401 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 5461 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 5521 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 5581 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 5641 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 5701 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 5761 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 5821 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 5881 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 5941 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 6001 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 6061 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 6121 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 6181 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 6241 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 6301 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 6361 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 6421 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 6481 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 6541 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 6601 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 6661 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 6721 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 6781 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 6841 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 6901 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 6961 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 7021 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 7081 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 7141 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg 7201 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat 7261 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg 7321 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc 7381 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa 7441 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg 7501 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa 7561 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt 7621 tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg 7681 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag 7741 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc 7801 cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt gttaaagaca 7861 ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag 7921 agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaa //