LOCUS Exported 3429 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Knock Down SH3+ Collybistin isoforms. ACCESSION . VERSION . KEYWORDS CBSH3+ shRNA-1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3429) AUTHORS de Blas, A TITLE de Blas Collybistin plasmids JOURNAL unpublished REFERENCE 2 (bases 1 to 3429) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3429 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 171..483 /label=U6 promoter /note="RNA polymerase III promoter for mouse U6 snRNA (Das et al., 1988)" primer_bind 402..422 /label=mU6-F /note="Mouse U6 promoter, forward primer" polyA_signal 551..685 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(575..594) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 629..648 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(783..803) /label=T3 /note="T3 promoter, forward primer" promoter complement(783..801) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(822..838) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(822..838) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(835..857) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 846..862 /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(870..900) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 915..936 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1053..1070) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1224..1812) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1304..1323) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1983..2843) /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 2606..2625 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2844..2948) /gene="bla" /label=AmpR promoter rep_origin complement(2974..3429) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3111..3132) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3323..3342 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc 61 tattacgcca gtcgacagcc tgcaggaatg gccggccatc ttaattaaga aatttaaata 121 tgtttaaaca tagggcccaa tggcgcgcca aggccatcta ggccaagctt atccgacgcc 181 gccatctcta ggcccgcgcc ggccccctcg cacagacttg tgggagaagc tcggctactc 241 ccctgccccg gttaatttgc atataatatt tcctagtaac tatagaggct taatgtgcga 301 taaaagacag ataatctgtt ctttttaata ctagctacat tttacatgat aggcttggat 361 ttctataaga gatacaaata ctaaattatt attttaaaaa acagcacaaa aggaaactca 421 ccctaactgt aaagtaattg tgtgttttga gactataaat atcccttgga gaaaagcctt 481 gtttgggatg cttccaacaa ggattgcgaa caatccttgt tggaagcatc ctttttctag 541 ataatacgta gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag 601 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac 661 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt 721 tcagggggag gtgtgggagg ttttttaatt cttgcggccg cggcggtacc cagcttttgt 781 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg 841 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa 901 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 961 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 1021 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 1081 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 1141 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 1201 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 1261 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 1321 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 1381 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 1441 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 1501 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 1561 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 1621 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 1681 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 1741 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 1801 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 1861 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 1921 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 1981 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 2041 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 2101 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 2161 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 2221 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 2281 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 2341 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 2401 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 2461 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 2521 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 2581 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 2641 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 2701 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 2761 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 2821 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 2881 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 2941 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa 3001 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca 3061 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga 3121 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc 3181 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc 3241 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc 3301 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg 3361 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac 3421 agggcgcgt //