LOCUS       sequence-183294-        7302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral gRNA plasmid targeting human NUP107.
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: NUP107 F8.2 gRNA
FEATURES             Location/Qualifiers
     source          1..7302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..85
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    202..223
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(238..243,244..261,262..268)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    276..292
                     /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     281..303
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     300..316
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     300..316
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     335..355
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        337..355
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        383..609
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             610..790
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    837..962
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1455..1688
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1873..1917
                     /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        2095..2343
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2095..2115
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2266..2285
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2499..2616
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2685..2896
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2922..3521
                     /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(2922..2941)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3418..3438
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3537..4125
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3590..3610)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4008..4019)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4197..4430
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4508..4629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4545..4564)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4599..4618
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4669..4804
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4824..4843)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4825..4843)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4853..4870)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4853..4869)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4862..4884)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5011..5466
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5098..5117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5308..5329
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5492..5596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5597..5665,5666..6457)
                     /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(5815..5834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6628..7216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7117..7136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
       61 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
      121 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
      181 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      241 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      301 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
      361 aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg
      421 gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc atgccgattg
      481 gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg tctgacatgg
      541 attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg cctagctcga
      601 tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
      661 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
      721 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
      781 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
      841 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
      901 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
      961 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
     1021 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
     1081 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
     1141 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
     1201 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
     1261 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct
     1321 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     1381 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     1441 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     1501 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     1561 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     1621 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     1681 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     1741 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     1801 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     1861 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     1921 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     1981 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     2041 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cagagagggc
     2101 ctatttccca tgattccttc atatttgcat atacgataca aggctgttag agagataatt
     2161 agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa
     2221 taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt
     2281 accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac
     2341 accgctctgc ctcccggatt acaggtttaa gagctatgct ggaaacagca tagcaagttt
     2401 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttgaa
     2461 ttctagatct tgagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
     2521 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2581 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     2641 gatccacttt ggcgccggct cgagtggctc cggtgcccgt cagtgggcag agcgcacatc
     2701 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
     2761 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     2821 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     2881 tgccgccaga acacaggtgt cgtgacgcgg gatccgccac catgaccgag tacaagccca
     2941 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     3001 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     3061 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     3121 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     3181 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     3241 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     3301 ccgtcggagt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     3361 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     3421 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     3481 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aacgcgttaa gtcgacaatc
     3541 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     3601 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     3661 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     3721 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     3781 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     3841 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     3901 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     3961 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     4021 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     4081 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga ctttaagacc
     4141 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     4201 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     4261 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     4321 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     4381 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtacgtatag
     4441 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     4501 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4561 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4621 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta
     4681 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4741 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4801 ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     4861 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     4921 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     4981 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     5041 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5101 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5161 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5221 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5281 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5341 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5401 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     5461 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5521 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5581 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5641 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5701 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5761 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5821 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5881 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5941 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6001 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6061 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6121 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6181 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6241 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6301 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6361 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6421 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6481 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6541 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6601 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6661 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6721 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6781 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6841 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6901 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6961 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7021 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7081 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7141 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7201 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7261 gccttttgct cacatgttct ttcctgcgtt atcccctgat tc
//
