LOCUS       sequence-363620-        7760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a Ncor1-targeting gRNA and Cre-recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7760)
  AUTHORS   Cai H, Chew SK, Li C, Tsai MK, Andrejka L, Murray CW, Hughes NW, 
            Shuldiner EG, Ashkin EL, Tang R, Hung KL, Chen LC, Lee SYC, Yousefi 
            M, Lin WY, Kunder CA, Cong L, McFarland CD, Petrov DA, Swanton C, 
            Winslow MM
  TITLE     A functional taxonomy of tumor suppression in oncogenic KRAS-driven 
            lung cancer.
  JOURNAL   Cancer Discov. 2021 Feb 19. pii: 2159-8290.CD-20-1325. doi: 
            10.1158/2159-8290.CD-20-1325.
  PUBMED    33608386
REFERENCE   2  (bases 1 to 7760)
  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"; journalName: "Cancer 
            Discov. 2021 Feb 19. pii: 2159-8290.CD-20-1325. doi: 
            10.1158/2159-8290.CD-20-1325."
COMMENT     Sequence Label: Lenti-sgNcor1#2/Cre
FEATURES             Location/Qualifiers
     source          1..7760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(128..147)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        319..698
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        700..898
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     853..873
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             916..1096
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1143..1268
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1765..1998
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2183..2227
                     /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"
     misc_feature    2525..2642
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2724..2972
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2724..2744
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2895..2914
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2994..3069
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3122..3621
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3157..3180)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3554..3573
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3625..3645
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3646..4674
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     misc_feature    4730..5318
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4783..4803)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5201..5212)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5321..5337)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5322..5338)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5847..6027
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6089..6677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6169..6188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6848..7639,7640..7708))
                     /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     7471..7490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7709..7760,1..53))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
       61 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
      121 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
      181 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
      241 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      301 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      361 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      421 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      481 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      541 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      601 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      661 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      721 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      781 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      841 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg
      901 cgttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
      961 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     1021 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     1081 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     1141 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     1201 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     1261 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     1321 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     1381 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     1441 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     1501 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     1561 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat
     1621 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     1681 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     1741 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     1801 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     1861 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     1921 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     1981 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     2041 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     2101 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     2161 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     2221 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     2281 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     2341 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     2401 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     2461 attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc
     2521 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     2581 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     2641 ttcgggttta ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctagc
     2701 cctgcaggaa ctgtttaaac tgtgagggcc tatttcccat gattccttca tatttgcata
     2761 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
     2821 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     2881 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     2941 ctttatatat cttgtggaaa ggaccaaaca ccgcaatatg aatgggctga tgggttttag
     3001 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     3061 agtcggtgct tttttgtttg gcgcgcctat ggcttaatta acatggcggc cgcattctac
     3121 cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg
     3181 ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg
     3241 cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc
     3301 aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca
     3361 cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt
     3421 tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg
     3481 ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc
     3541 tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct
     3601 tcctcatctc cgggacccgg gatgcccaag aagaagagga aggtgtccaa tttactgacc
     3661 gtacaccaaa atttgcctgc attaccggtc gatgcaacga gtgatgaggt tcgcaagaac
     3721 ctgatggaca tgttcaggga tcgccaggcg ttttctgagc atacctggaa aatgcttctg
     3781 tccgtttgcc ggtcgtgggc ggcatggtgc aagttgaata accggaaatg gtttcccgca
     3841 gaacctgaag atgttcgcga ttatcttcta tatcttcagg cgcgcggtct ggcagtaaaa
     3901 actatccagc aacatttggg ccagctaaac atgcttcatc gtcggtccgg gctgccacga
     3961 ccaagtgaca gcaatgctgt ttcactggtt atgcggcgga tccgaaaaga aaacgttgat
     4021 gccggtgaac gtgcaaaaca ggctctagcg ttcgaacgca ctgatttcga ccaggttcgt
     4081 tcactcatgg aaaatagcga tcgctgccag gatatacgta atctggcatt tctggggatt
     4141 gcttataaca ccctgttacg tatagccgaa attgccagga tcagggttaa agatatctca
     4201 cgtactgacg gtgggagaat gttaatccat attggcagaa cgaaaacgct ggttagcacc
     4261 gcaggtgtag agaaggcact tagcctgggg gtaactaaac tggtcgagcg atggatttcc
     4321 gtctctggtg tagctgatga tccgaataac tacctgtttt gccgggtcag aaaaaatggt
     4381 gttgccgcgc catctgccac cagccagcta tcaactcgcg ccctggaagg gatttttgaa
     4441 gcaactcatc gattgattta cggcgctaag gatgactctg gtcagagata cctggcctgg
     4501 tctggacaca gtgcccgtgt cggagccgcg cgagatatgg cccgcgctgg agtttcaata
     4561 ccggagatca tgcaagctgg tggctggacc aatgtaaata ttgtcatgaa ctatatccgt
     4621 aacctggata gtgaaacagg ggcaatggtg cgcctgctgg aagatggcga ttagccatgt
     4681 ttaagggttc cggttccact aggtacaatt cgatatcaag cttatcgata atcaacctct
     4741 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     4801 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     4861 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     4921 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     4981 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     5041 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     5101 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     5161 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     5221 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     5281 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgatc
     5341 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt
     5401 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac
     5461 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     5521 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct
     5581 accacacaca aggctacttc cctgattggc agaactacac accagggcca gggatcagat
     5641 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag aaggtagaag
     5701 aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat gggatggatg
     5761 acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt catcacatgg
     5821 cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc tgagcctggg
     5881 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     5941 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6001 tttagtcagt gtggaaaatc tctagcagca tgtgagcaaa aggccagcaa aaggccagga
     6061 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6121 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6181 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6241 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6301 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6361 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6421 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6481 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6541 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6601 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6661 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6721 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6781 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6841 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6901 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6961 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7021 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7081 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7141 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7201 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7261 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7321 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7381 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7441 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7501 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7561 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7621 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7681 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7741 atcagggtta ttgtctcatg
//
