LOCUS       sequence-363636-        7759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a Lats1-targeting gRNA and Cre-recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7759)
  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 7759)
  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-sgLats1#2/Cre
FEATURES             Location/Qualifiers
     source          1..7759
                     /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        2993..3068
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3121..3620
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3156..3179)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3553..3572
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3624..3644
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3645..4673
                     /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    4729..5317
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4782..4802)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5200..5211)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5320..5336)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5321..5337)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5846..6026
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6088..6676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6168..6187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6847..7638,7639..7707))
                     /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     7470..7489
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7708..7759,1..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 ccgcagacat ctgctctcga cggttttaga
     3001 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     3061 gtcggtgctt ttttgtttgg cgcgcctatg gcttaattaa catggcggcc gcattctacc
     3121 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
     3181 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
     3241 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
     3301 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
     3361 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
     3421 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
     3481 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
     3541 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
     3601 cctcatctcc gggacccggg atgcccaaga agaagaggaa ggtgtccaat ttactgaccg
     3661 tacaccaaaa tttgcctgca ttaccggtcg atgcaacgag tgatgaggtt cgcaagaacc
     3721 tgatggacat gttcagggat cgccaggcgt tttctgagca tacctggaaa atgcttctgt
     3781 ccgtttgccg gtcgtgggcg gcatggtgca agttgaataa ccggaaatgg tttcccgcag
     3841 aacctgaaga tgttcgcgat tatcttctat atcttcaggc gcgcggtctg gcagtaaaaa
     3901 ctatccagca acatttgggc cagctaaaca tgcttcatcg tcggtccggg ctgccacgac
     3961 caagtgacag caatgctgtt tcactggtta tgcggcggat ccgaaaagaa aacgttgatg
     4021 ccggtgaacg tgcaaaacag gctctagcgt tcgaacgcac tgatttcgac caggttcgtt
     4081 cactcatgga aaatagcgat cgctgccagg atatacgtaa tctggcattt ctggggattg
     4141 cttataacac cctgttacgt atagccgaaa ttgccaggat cagggttaaa gatatctcac
     4201 gtactgacgg tgggagaatg ttaatccata ttggcagaac gaaaacgctg gttagcaccg
     4261 caggtgtaga gaaggcactt agcctggggg taactaaact ggtcgagcga tggatttccg
     4321 tctctggtgt agctgatgat ccgaataact acctgttttg ccgggtcaga aaaaatggtg
     4381 ttgccgcgcc atctgccacc agccagctat caactcgcgc cctggaaggg atttttgaag
     4441 caactcatcg attgatttac ggcgctaagg atgactctgg tcagagatac ctggcctggt
     4501 ctggacacag tgcccgtgtc ggagccgcgc gagatatggc ccgcgctgga gtttcaatac
     4561 cggagatcat gcaagctggt ggctggacca atgtaaatat tgtcatgaac tatatccgta
     4621 acctggatag tgaaacaggg gcaatggtgc gcctgctgga agatggcgat tagccatgtt
     4681 taagggttcc ggttccacta ggtacaattc gatatcaagc ttatcgataa tcaacctctg
     4741 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     4801 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     4861 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4921 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4981 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     5041 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     5101 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     5161 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     5221 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     5281 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgatcg
     5341 agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg
     5401 tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc
     5461 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     5521 gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta
     5581 ccacacacaa ggctacttcc ctgattggca gaactacaca ccagggccag ggatcagata
     5641 tccactgacc tttggatggt gctacaagct agtaccagtt gagcaagaga aggtagaaga
     5701 agccaatgaa ggagagaaca cccgcttgtt acaccctgtg agcctgcatg ggatggatga
     5761 cccggagaga gaagtattag agtggaggtt tgacagccgc ctagcatttc atcacatggc
     5821 ccgagagctg catccggact gtactgggtc tctctggtta gaccagatct gagcctggga
     5881 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     5941 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6001 ttagtcagtg tggaaaatct ctagcagcat gtgagcaaaa ggccagcaaa aggccaggaa
     6061 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6121 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6181 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6241 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6301 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6361 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6421 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6481 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6541 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6601 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6661 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6721 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6781 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6841 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6901 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6961 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7021 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7081 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7141 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7201 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7261 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7321 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7381 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7441 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7501 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7561 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7621 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7681 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7741 tcagggttat tgtctcatg
//
