LOCUS Exported 5825 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Provides negative selection cassette on the vector. ACCESSION . VERSION . KEYWORDS pDEST-R4R3-MC1DTA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5825) TITLE Minoru Ko Lab plasmids REFERENCE 2 (bases 1 to 5825) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5825 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..17 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 1..17 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" protein_bind 36..160 /gene="mutant version of attR" /label=attR4 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS complement(508..813) /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" primer_bind complement(610..629) /label=ccdB-fwd /note="ccdB gene, forward primer" CDS complement(1163..1816) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG" primer_bind 1731..1750 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(1817..1919) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" protein_bind complement(2065..2188) /gene="mutant version of attR" /label=attR3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2196..2213) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2196..2212) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2205..2227) /label=M13/pUC Forward /note="In lacZ gene" CDS 2663..3235 /codon_start=1 /gene="Corynebacterium diphtheriae tox" /product="diphtheria toxin fragment A" /label=DTA /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI NFETRGKRGQDAMYEYMAQACAGNRVRR" intron 3312..3377 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" primer_bind 3810..3832 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3870..3888) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3956..4060 /gene="bla" /label=AmpR promoter CDS 4061..4921 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(4279..4298) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5092..5680 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5581..5600 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5807..4 /label=M13/pUC Reverse /note="In lacZ gene" ORIGIN 1 caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga aaagttgaac 61 gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga 121 ctacataata ctgtaaaaca caacatatcc agtcactatg gtcgaccgac agccttccaa 181 atgttcttct caaacggaat cgtcgtatcc agcctactcg ctattgtcct caatgccgta 241 ttaaatcata aaaagaaata agaaaaagag gtgcgagcct cttttttgtg tgacaaaata 301 aaaacatcta cctattcata tacgctagtg tcatagtcct gaaaatcatc tgcatcaaga 361 acaatttcac aactcttata cttttctctt acaagtcgtt cggcttcatc tggattttca 421 gcctctatac ttactaaacg tgataaagtt tctgtaattt ctactgtatc gacctgcaga 481 ctggctgtgt ataagggagc ctgacattta tattccccag aacatcaggt taatggcgtt 541 tttgatgtca ttttcgcggt ggctgagatc agccacttct tccccgataa cggagaccgg 601 cacactggcc atatcggtgg tcatcatgcg ccagctttca tccccgatat gcaccaccgg 661 gtaaagttca cgggagactt tatctgacag cagacgtgca ctggccaggg ggatcaccat 721 ccgtcgcccg ggcgtgtcaa taatatcact ctgtacatcc acaaacagac gataacggct 781 ctctctttta taggtgtaaa ccttaaactg catttcacca gcccctgttc tcgtcagcaa 841 aagagccgtt catttcaata aaccgggcga cctcagccat cccttcctga ttttccgctt 901 tccagcgttc ggcacgcaga cgacgggctt cattctgcat ggttgtgctt accagaccgg 961 agatattgac atcatatatg ccttgagcaa ctgatagctg tcgctgtcaa ctgtcactgt 1021 aatacgctgc ttcatagcat acctcttttt gacatacttc gggtatacat atcagtatat 1081 attcttatac cgcaaaaatc agcgcgcaaa tacgcatact gttatctggc ttttagtaag 1141 ccggatccac gcggcgttta cgccccccct gccactcatc gcagtactgt tgtaattcat 1201 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg 1261 gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga 1321 agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg 1381 agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac 1441 acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc 1501 agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat 1561 cccatatcac cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca 1621 ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct 1681 ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact 1741 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag 1801 tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata 1861 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa 1921 cgtctcattt tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat 1981 ttatttattc tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg 2041 gtgatgctgc caacttacgg ccgccatagt gactggatat gttgtgtttt acagtattat 2101 gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt 2161 tctcgttcaa ctttattata catagttgat aattcactgg ccgtcgtttt acaacgtcgt 2221 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc 2281 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg 2341 aatggcgaat ggcgccgctt atcgataccg tcgagcagtg tggttttcaa gaggaagcaa 2401 aaagcctctc cacccaggcc tggaatgttt ccacccaatg tcgagcagtg tgattttgca 2461 agaggaagca aaaagcctct ccacccaggc ctggaatgtt tccacccaat gtcgagcaaa 2521 ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg gggcggcgcg 2581 gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc gagcgaccct 2641 gcaggtcctc gccatggatc ctgatgatgt tgttgattct tctaaatctt ttgtgatgga 2701 aaacttttct tcgtaccacg ggactaaacc tggttatgta gattccattc aaaaaggtat 2761 acaaaagcca aaatctggta cacaaggaaa ttatgacgat gattggaaag ggttttatag 2821 taccgacaat aaatacgacg ctgcgggata ctctgtagat aatgaaaacc cgctctctgg 2881 aaaagctgga ggcgtggtca aagtgacgta tccaggactg acgaaggttc tcgcactaaa 2941 agtggataat gccgaaacta ttaagaaaga gttaggttta agtctcactg aaccgttgat 3001 ggagcaagtc ggaacggaag agtttatcaa aaggttcggt gatggtgctt cgcgtgtagt 3061 gctcagcctt cccttcgctg aggggagttc tagcgttgaa tatattaata actgggaaca 3121 ggcgaaagcg ttaagcgtag aacttgagat taattttgaa acccgtggaa aacgtggcca 3181 agatgcgatg tatgagtata tggctcaagc ctgtgcagga aatcgtgtca ggcgatctct 3241 ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct acagagattt 3301 aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa 3361 ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag tggtggaatg 3421 cagatccact agagtcgagg gatgcagaaa ttgatgatct attaaacaat aaagatgtcc 3481 actaaaatgg aagtttttcc tgtcatactt tgttaagaag ggtgagaaca gagtacctac 3541 attttgaatg gaaggattgg agctacgggg gtgggggtgg ggtgggatta gataaatgcc 3601 tgctctttac tgaaggctct ttactattgc tttatgataa tgtttcatag ttggatatca 3661 taatttaaac aacatatggt gcactctcag tgcaatctgc tctgatgccg catagttaag 3721 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc 3781 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc 3841 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa 3901 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg 3961 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata 4021 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg 4081 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac 4141 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact 4201 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat 4261 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga 4321 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac 4381 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat 4441 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac 4501 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct 4561 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac 4621 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga 4681 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg 4741 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact 4801 ggggtcagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac 4861 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta 4921 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt 4981 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 5041 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 5101 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 5161 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 5221 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc 5281 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 5341 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 5401 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 5461 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 5521 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 5581 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 5641 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 5701 tttacggttc ctggtctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 5761 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccagcg gataacaatt 5821 tcaca //