LOCUS Exported 3429 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Knock Down all Collybistin isoforms. ACCESSION . VERSION . KEYWORDS AllCB 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synthetic DNA construct REFERENCE 1 (bases 1 to 3429) AUTHORS de Blas, A TITLE de Blas Collybistin plasmids JOURNAL unpublished REFERENCE 2 (bases 1 to 3429) AUTHORS .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" primer_bind complement(30..47) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(201..789) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(281..300) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(960..1820) /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 1583..1602 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1821..1925) /gene="bla" /label=AmpR promoter rep_origin complement(1951..2406) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2088..2109) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2300..2319 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2577..2889 /label=U6 promoter /note="RNA polymerase III promoter for mouse U6 snRNA (Das et al., 1988)" primer_bind 2808..2828 /label=mU6-F /note="Mouse U6 promoter, forward primer" polyA_signal 2957..3091 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2981..3000) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3035..3054 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3189..3209) /label=T3 /note="T3 promoter, forward primer" promoter complement(3189..3207) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3228..3244) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3228..3244) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3241..3263) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3252..3268 /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(3276..3306) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3321..3342 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 61 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 361 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 601 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 841 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 901 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 961 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 1021 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 1081 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 1141 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 1201 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 1261 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 1321 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 1381 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 1441 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 1501 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 1561 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 1621 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 1681 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 1741 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 1801 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 1861 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 1921 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat 1981 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa 2041 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca 2101 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg 2161 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta 2221 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg 2281 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa 2341 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg 2401 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct 2461 cttcgctatt acgccagtcg acagcctgca ggaatggccg gccatcttaa ttaagaaatt 2521 taaatatgtt taaacatagg gcccaatggc gcgccaaggc catctaggcc aagcttatcc 2581 gacgccgcca tctctaggcc cgcgccggcc ccctcgcaca gacttgtggg agaagctcgg 2641 ctactcccct gccccggtta atttgcatat aatatttcct agtaactata gaggcttaat 2701 gtgcgataaa agacagataa tctgttcttt ttaatactag ctacatttta catgataggc 2761 ttggatttct ataagagata caaatactaa attattattt taaaaaacag cacaaaagga 2821 aactcaccct aactgtaaag taattgtgtg ttttgagact ataaatatcc cttggagaaa 2881 agccttgttt gggatgcttc caacaaggat tgcgaacaat ccttgttgga agcatccttt 2941 ttctagataa tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac 3001 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt 3061 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt 3121 tcaggttcag ggggaggtgt gggaggtttt ttaattcttg cggccgcggc ggtacccagc 3181 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt 3241 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg gagcataaag 3301 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 3361 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 3421 gggagaggc //