LOCUS Exported 5586 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Enhanced RNAi vektor for C. elegans. Contains two T7 polymerase termination sequences adjacent to the T7 promoter sites and a lacZ coding sequence.. ACCESSION . VERSION . KEYWORDS T777T SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5586) AUTHORS Sturm A, Saskoi E, Tibor K, Weinhardt N, Vellai T TITLE Highly efficient RNAi and Cas9-based auto-cloning systems for C. elegans research. JOURNAL Nucleic Acids Res. 2018 Jun 19. pii: 5040046. doi: 10.1093/nar/gky516. PUBMED 29924347 REFERENCE 2 (bases 1 to 5586) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5586 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 2..49 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter 50..68 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 93..109 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 94..110 /label=KS primer /note="common sequencing primer, one of multiple similar variants" CDS 199..3246 /codon_start=1 /gene="lacZ" /product="beta-galactosidase" /label=lacZ /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD SWSPSVSAEFQLSAGRYHYQLVWCQK" primer_bind complement(201..223) /label=M13/pUC Forward /note="In lacZ gene" promoter complement(3379..3397) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" terminator 3398..3445 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin complement(3507..4095) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3587..3606) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4266..5126) /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" primer_bind 4889..4908 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5127..5231) /gene="bla" /label=AmpR promoter primer_bind complement(5394..5415) /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 ccaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt aatacgactc 61 actatagggc gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttag 121 tctccgaagg aacgtgtcac ggttgacagc agtcctaggt ataatactag tggactttaa 181 gaaggagata tacatatggt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc 241 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc 301 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg 361 tttccggcac cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat 421 actgtcgtcg tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac 481 gtgacctatc ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt 541 tactcgctca catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt 601 tttgatggcg ttaactcggc gtttcatctg tggtgcaacg ggcgctgggt cggttacggc 661 caggacagtc gtttgccgtc tgaatttgac ctgagcgcat ttttacgcgc cggagaaaac 721 cgcctcgcgg tgatggtgct gcgctggagt gacggcagtt atctggaaga tcaggatatg 781 tggcggatga gcggcatttt ccgtgacgtc tcgttgctgc ataaaccgac tacacaaatc 841 agcgatttcc atgttgccac tcgctttaat gatgatttca gccgcgctgt actggaggct 901 gaagttcaga tgtgcggcga gttgcgtgac tacctacggg taacagtttc tttatggcag 961 ggtgaaacgc aggtcgccag cggcaccgcg cctttcggcg gtgaaattat cgatgagcgt 1021 ggtggttatg ccgatcgcgt cacactacgt ctgaacgtcg aaaacccgaa actgtggagc 1081 gccgaaatcc cgaatctcta tcgtgcggtg gttgaactgc acaccgccga cggcacgctg 1141 attgaagcag aagcctgcga tgtcggtttc cgcgaggtgc ggattgaaaa tggtctgctg 1201 ctgctgaacg gcaagccgtt gctgattcga ggcgttaacc gtcacgagca tcatcctctg 1261 catggtcagg tcatggatga gcagacgatg gtgcaggata tcctgctgat gaagcagaac 1321 aactttaacg ccgtgcgctg ttcgcattat ccgaaccatc cgctgtggta cacgctgtgc 1381 gaccgctacg gcctgtatgt ggtggatgaa gccaatattg aaacccacgg catggtgcca 1441 atgaatcgtc tgaccgatga tccgcgctgg ctaccggcga tgagcgaacg cgtaacgcga 1501 atggtgcagc gcgatcgtaa tcacccgagt gtgatcatct ggtcgctggg gaatgaatca 1561 ggccacggcg ctaatcacga cgcgctgtat cgctggatca aatctgtcga tccttcccgc 1621 ccggtgcagt atgaaggcgg cggagccgac accacggcca ccgatattat ttgcccgatg 1681 tacgcgcgcg tggatgaaga ccagcccttc ccggctgtgc cgaaatggtc catcaaaaaa 1741 tggctttcgc tacctggaga gacgcgcccg ctgatccttt gcgaatacgc ccacgcgatg 1801 ggtaacagtc ttggcggttt cgctaaatac tggcaggcgt ttcgtcagta tccccgttta 1861 cagggcggct tcgtctggga ctgggtggat cagtcgctga ttaaatatga tgaaaacggc 1921 aacccgtggt cggcttacgg cggtgatttt ggcgatacgc cgaacgatcg ccagttctgt 1981 atgaacggtc tggtctttgc cgaccgcacg ccgcatccag cgctgacgga agcaaaacac 2041 cagcagcagt ttttccagtt ccgtttatcc gggcaaacca tcgaagtgac cagcgaatac 2101 ctgttccgtc atagcgataa cgagctcctg cactggatgg tggcgctgga tggtaagccg 2161 ctggcaagcg gtgaagtgcc tctggatgtc gctccacaag gtaaacagtt gattgaactg 2221 cctgaactac cgcagccgga gagcgccggg caactctggc tcacagtacg cgtagtgcaa 2281 ccgaacgcga ccgcatggtc agaagccggg cacatcagcg cctggcagca gtggcgtctg 2341 gcggaaaacc tcagtgtgac gctccccgcc gcgtcccacg ccatcccgca tctgaccacc 2401 agcgaaatgg atttttgcat cgagctgggt aataagcgtt ggcaatttaa ccgccagtca 2461 ggctttcttt cacagatgtg gattggcgat aaaaaacaac tgctgacgcc gctgcgcgat 2521 cagttcaccc gtgcaccgct ggataacgac attggcgtaa gtgaagcgac ccgcattgac 2581 cctaacgcct gggtcgaacg ctggaaggcg gcgggccatt accaggccga agcagcgttg 2641 ttgcagtgca cggcagatac acttgctgat gcggtgctga ttacgaccgc tcacgcgtgg 2701 cagcatcagg ggaaaacctt atttatcagc cggaaaacct accggattga tggtagtggt 2761 caaatggcga ttaccgttga tgttgaagtg gcgagcgata caccgcatcc ggcgcggatt 2821 ggcctgaact gccagctggc gcaggtagca gagcgggtaa actggctcgg attagggccg 2881 caagaaaact atcccgaccg ccttactgcc gcctgttttg accgctggga tctgccattg 2941 tcagacatgt ataccccgta cgtcttcccg agcgaaaacg gtctgcgctg cgggacgcgc 3001 gaattgaatt atggcccaca ccagtggcgc ggcgacttcc agttcaacat cagccgctac 3061 agtcaacagc aactgatgga aaccagccat cgccatctgc tgcacgcgga agaaggcaca 3121 tggctgaata tcgacggttt ccatatgggg attggtggcg acgactcctg gagcccgtca 3181 gtatcggcgg aattccagct gagcgccggt cgctaccatt accagttggt ctggtgtcaa 3241 aaataataac ccttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttccgtgaca 3301 cgttccttcg gagactctag agcggccgcc accgcggtgg agctcgaatt catcgatgat 3361 atcagatctg ccggtctccc tatagtgagt cgtattacta gcataacccc ttggggcctc 3421 taaacgggtc ttgaggggtt ttttgacatg tgagcaaaag gccagcaaaa ggccaggaac 3481 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 3541 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 3601 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 3661 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 3721 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 3781 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 3841 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 3901 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt 3961 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 4021 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 4081 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 4141 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 4201 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 4261 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 4321 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 4381 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 4441 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 4501 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 4561 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 4621 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 4681 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 4741 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 4801 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 4861 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 4921 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 4981 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 5041 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 5101 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 5161 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 5221 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt 5281 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg 5341 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt 5401 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct 5461 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt 5521 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa 5581 agccgg //