LOCUS       Exported                3427 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 3427)
  AUTHORS   de Blas, A
  TITLE     de Blas Collybistin plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 3427)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3427
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(36..896)
                     /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     659..678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(897..1001)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1027..1482)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1164..1185)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1376..1395
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1653..1965
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     1884..1904
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     polyA_signal    2031..2165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2055..2074)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2109..2128
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2263..2283)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2263..2281)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2302..2318)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2302..2318)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2315..2337)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2326..2342
                     /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(2350..2380)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2395..2416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2533..2550)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2704..3292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2784..2803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
       61 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
      121 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
      181 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
      241 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
      301 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
      361 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
      421 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
      481 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
      541 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
      601 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
      661 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
      721 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
      781 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
      841 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
      901 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
      961 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1021 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     1081 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     1141 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     1201 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     1261 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     1321 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     1381 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     1441 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     1501 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgacag
     1561 cctgcaggaa tggccggcca tcttaattaa gaaatttaaa tatgtttaaa catagggccc
     1621 aatggcgcgc caaggccatc taggccaagc ttatccgacg ccgccatctc taggcccgcg
     1681 ccggccccct cgcacagact tgtgggagaa gctcggctac tcccctgccc cggttaattt
     1741 gcatataata tttcctagta actatagagg cttaatgtgc gataaaagac agataatctg
     1801 ttctttttaa tactagctac attttacatg ataggcttgg atttctataa gagatacaaa
     1861 tactaaatta ttattttaaa aaacagcaca aaaggaaact caccctaact gtaaagtaat
     1921 tgtgtgtttt gagactataa atatcccttg gagaaaagcc ttgtttgcgc attgacatgg
     1981 ataaatacga atatttatcc atgtcaatgc gtttttctag ataatacgta gatccagaca
     2041 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2101 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2161 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2221 ttttttaatt cttgcggccg cggcggtacc cagcttttgt tccctttagt gagggttaat
     2281 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2341 aattccacac aacatacgag ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt
     2401 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2461 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3121 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3421 taaatca
//
