LOCUS       sequence-183350-        7302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human DYNLL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7302)
  AUTHORS   McKinley KL, Cheeseman IM
  TITLE     Large-Scale Analysis of CRISPR/Cas9 Cell-Cycle Knockouts Reveals the
            Diversity of p53-Dependent Responses to Cell-Cycle Defects.
  JOURNAL   Dev Cell. 2017 Feb 27;40(4):405-420.e2. doi: 
            10.1016/j.devcel.2017.01.012. Epub 2017 Feb 16.
  PUBMED    28216383
REFERENCE   2  (bases 1 to 7302)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.devcel.2017.01.012"; journalName: "Dev Cell"; date: 
            "2017-02-27- 27"; volume: "40"; issue: "4"; pages: "405-420.e2"
COMMENT     Sequence Label: DYNLL1 A7.5 gRNA
FEATURES             Location/Qualifiers
     source          1..7302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    111..132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(147..152,153..170,171..177)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    185..201
                     /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)."
     primer_bind     190..212
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     209..225
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     209..225
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     244..264
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        246..264
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        292..518
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             519..699
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    746..871
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1364..1597
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1782..1826
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        2004..2252
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2004..2024
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2175..2194
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2408..2525
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2594..2805
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2831..3430
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2831..2850)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3327..3347
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3446..4034
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3499..3519)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3917..3928)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4106..4339
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4417..4538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4454..4473)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4508..4527
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4578..4713
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4733..4752)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4734..4752)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4762..4779)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4762..4778)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4771..4793)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4920..5375
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5007..5026)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5217..5238
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5401..5505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5506..5574,5575..6366)
                     /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     complement(5724..5743)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6537..7125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7026..7045
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7279..7296
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
       61 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
      121 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
      181 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
      241 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
      301 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
      361 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
      421 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
      481 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
      541 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
      601 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
      661 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
      721 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
      781 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
      841 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
      901 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
      961 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     1021 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     1081 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     1141 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     1201 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     1261 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     1321 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     1381 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     1441 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     1501 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     1561 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     1621 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     1681 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     1741 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     1801 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     1861 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     1921 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     1981 tcccaacccc gaggggaccc agagagggcc tatttcccat gattccttca tatttgcata
     2041 tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat
     2101 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     2161 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     2221 ctttatatat cttgtggaaa ggacgaaaca ccgtcccacc tggcattgca tcggtttaag
     2281 agctatgctg gaaacagcat agcaagttta aataaggcta gtccgttatc aacttgaaaa
     2341 agtggcaccg agtcggtgct ttttttgaat tctagatctt gagacaaatg gcagtattca
     2401 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
     2461 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     2521 attttcgggt ttattacagg gacagcagag atccactttg gcgccggctc gagtggctcc
     2581 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     2641 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     2701 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     2761 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtgtc gtgacgcggg
     2821 atccgccacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     2881 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     2941 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     3001 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     3061 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     3121 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     3181 gcccaaggag cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc accagggcaa
     3241 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     3301 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     3361 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     3421 cggtgcctga acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     3481 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3541 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3601 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3661 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3721 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3781 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3841 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3901 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3961 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     4021 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
     4081 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     4141 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     4201 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     4261 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     4321 gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta
     4381 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     4441 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4501 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     4561 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4621 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4681 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     4741 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     4801 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4861 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     4921 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4981 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5041 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5101 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5161 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5221 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5281 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5341 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     5401 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5461 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5521 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5581 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5641 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5701 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5761 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5821 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5881 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5941 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6001 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6061 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6121 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6181 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6241 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6301 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6361 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6421 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6481 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6541 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6601 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6661 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6721 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6781 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6841 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6901 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6961 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7021 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7081 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7141 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7201 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7261 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cg
//
