LOCUS       sequence-363553-        7759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a Ptprd-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-sgPtprd#2/Cre
FEATURES             Location/Qualifiers
     source          1..7759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(55..74)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        246..625
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        627..825
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     780..800
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             843..1023
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1070..1195
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1692..1925
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2110..2154
                     /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    2452..2569
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2651..2899
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2651..2671
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2822..2841
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2920..2995
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3048..3547
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3083..3106)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3480..3499
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3551..3571
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3572..4600
                     /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    4656..5244
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4709..4729)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5127..5138)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5247..5263)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5248..5264)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5773..5953
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6015..6603)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6095..6114)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6774..7565,7566..7634))
                     /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     7397..7416
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7635..7739)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
       61 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
      121 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
      181 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      241 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      301 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      361 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      421 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      481 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      541 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      601 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      661 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      721 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      781 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta
      841 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
      901 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
      961 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     1021 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
     1081 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     1141 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     1201 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     1261 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     1321 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     1381 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     1441 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     1501 gcaaaacaaa agtaagacca ccgcacagca agcggccggc cgctgatctt cagacctgga
     1561 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     1621 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     1681 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     1741 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     1801 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     1861 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     1921 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     1981 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     2041 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     2101 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     2161 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     2221 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     2281 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
     2341 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
     2401 tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag
     2461 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     2521 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     2581 cagggacagc agagatccag tttggttagt accgggcccg ctctagccct gcaggaactg
     2641 tttaaactgt gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
     2701 tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
     2761 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
     2821 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
     2881 gtggaaagga ccaaacaccg caatggtatc tgatcgtggg ttttagagct agaaatagca
     2941 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     3001 tgtttggcgc gcctatggct taattaacat ggcggccgca ttctaccggg taggggaggc
     3061 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
     3121 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
     3181 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
     3241 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
     3301 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
     3361 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
     3421 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
     3481 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
     3541 acccgggatg cccaagaaga agaggaaggt gtccaattta ctgaccgtac accaaaattt
     3601 gcctgcatta ccggtcgatg caacgagtga tgaggttcgc aagaacctga tggacatgtt
     3661 cagggatcgc caggcgtttt ctgagcatac ctggaaaatg cttctgtccg tttgccggtc
     3721 gtgggcggca tggtgcaagt tgaataaccg gaaatggttt cccgcagaac ctgaagatgt
     3781 tcgcgattat cttctatatc ttcaggcgcg cggtctggca gtaaaaacta tccagcaaca
     3841 tttgggccag ctaaacatgc ttcatcgtcg gtccgggctg ccacgaccaa gtgacagcaa
     3901 tgctgtttca ctggttatgc ggcggatccg aaaagaaaac gttgatgccg gtgaacgtgc
     3961 aaaacaggct ctagcgttcg aacgcactga tttcgaccag gttcgttcac tcatggaaaa
     4021 tagcgatcgc tgccaggata tacgtaatct ggcatttctg gggattgctt ataacaccct
     4081 gttacgtata gccgaaattg ccaggatcag ggttaaagat atctcacgta ctgacggtgg
     4141 gagaatgtta atccatattg gcagaacgaa aacgctggtt agcaccgcag gtgtagagaa
     4201 ggcacttagc ctgggggtaa ctaaactggt cgagcgatgg atttccgtct ctggtgtagc
     4261 tgatgatccg aataactacc tgttttgccg ggtcagaaaa aatggtgttg ccgcgccatc
     4321 tgccaccagc cagctatcaa ctcgcgccct ggaagggatt tttgaagcaa ctcatcgatt
     4381 gatttacggc gctaaggatg actctggtca gagatacctg gcctggtctg gacacagtgc
     4441 ccgtgtcgga gccgcgcgag atatggcccg cgctggagtt tcaataccgg agatcatgca
     4501 agctggtggc tggaccaatg taaatattgt catgaactat atccgtaacc tggatagtga
     4561 aacaggggca atggtgcgcc tgctggaaga tggcgattag ccatgtttaa gggttccggt
     4621 tccactaggt acaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     4681 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     4741 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     4801 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     4861 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     4921 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     4981 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     5041 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     5101 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     5161 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     5221 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgatcgaga cctagaaaaa
     5281 catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa
     5341 gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg
     5401 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     5461 ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc
     5521 tacttccctg attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt
     5581 ggatggtgct acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga
     5641 gagaacaccc gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa
     5701 gtattagagt ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat
     5761 ccggactgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     5821 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     5881 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     5941 aaaatctcta gcagcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6001 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6061 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6121 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6181 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6241 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6301 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6361 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6421 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6481 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6541 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6601 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6661 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6721 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6781 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6841 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6901 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6961 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7021 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7081 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7141 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7201 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7261 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7321 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7381 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7441 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7501 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7561 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7621 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7681 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7741 acatttcccc gaaaagtgc
//
