LOCUS       sequence-361935-        7760 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 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-sgPtprd#1/Cre
FEATURES             Location/Qualifiers
     source          1..7760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(166..185)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        357..736
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        738..936
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     891..911
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             954..1134
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1181..1306
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1803..2036
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2221..2265
                     /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    2563..2680
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2762..3010
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2762..2782
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2933..2952
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3032..3107
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3160..3659
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3195..3218)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3592..3611
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3663..3683
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3684..4712
                     /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    4768..5356
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4821..4841)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5239..5250)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5359..5375)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5360..5376)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5885..6065
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6127..6715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6207..6226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6886..7677,7678..7746))
                     /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     7509..7528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7747..7760,1..91))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
      181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      361 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      421 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      481 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      541 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      601 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      661 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      721 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      781 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      841 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      901 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      961 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     1021 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     1081 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1141 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1201 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1261 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1321 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1381 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1441 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1501 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1561 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1621 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     1681 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1741 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1801 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1861 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1921 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1981 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     2041 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     2101 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     2161 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2221 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2281 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2341 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2401 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2461 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2521 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2581 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2641 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2701 cagagatcca gtttggttag taccgggccc gctctagccc tgcaggaact gtttaaactg
     2761 tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
     2821 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
     2881 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
     2941 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
     3001 accaaacacc gcttggtgcg gagcacatct ggttttagag ctagaaatag caagttaaaa
     3061 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgtttggc
     3121 gcgcctatgg cttaattaac atggcggccg cattctaccg ggtaggggag gcgcttttcc
     3181 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg
     3241 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg
     3301 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc
     3361 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga
     3421 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc
     3481 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca
     3541 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
     3601 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggacccggga
     3661 tgcccaagaa gaagaggaag gtgtccaatt tactgaccgt acaccaaaat ttgcctgcat
     3721 taccggtcga tgcaacgagt gatgaggttc gcaagaacct gatggacatg ttcagggatc
     3781 gccaggcgtt ttctgagcat acctggaaaa tgcttctgtc cgtttgccgg tcgtgggcgg
     3841 catggtgcaa gttgaataac cggaaatggt ttcccgcaga acctgaagat gttcgcgatt
     3901 atcttctata tcttcaggcg cgcggtctgg cagtaaaaac tatccagcaa catttgggcc
     3961 agctaaacat gcttcatcgt cggtccgggc tgccacgacc aagtgacagc aatgctgttt
     4021 cactggttat gcggcggatc cgaaaagaaa acgttgatgc cggtgaacgt gcaaaacagg
     4081 ctctagcgtt cgaacgcact gatttcgacc aggttcgttc actcatggaa aatagcgatc
     4141 gctgccagga tatacgtaat ctggcatttc tggggattgc ttataacacc ctgttacgta
     4201 tagccgaaat tgccaggatc agggttaaag atatctcacg tactgacggt gggagaatgt
     4261 taatccatat tggcagaacg aaaacgctgg ttagcaccgc aggtgtagag aaggcactta
     4321 gcctgggggt aactaaactg gtcgagcgat ggatttccgt ctctggtgta gctgatgatc
     4381 cgaataacta cctgttttgc cgggtcagaa aaaatggtgt tgccgcgcca tctgccacca
     4441 gccagctatc aactcgcgcc ctggaaggga tttttgaagc aactcatcga ttgatttacg
     4501 gcgctaagga tgactctggt cagagatacc tggcctggtc tggacacagt gcccgtgtcg
     4561 gagccgcgcg agatatggcc cgcgctggag tttcaatacc ggagatcatg caagctggtg
     4621 gctggaccaa tgtaaatatt gtcatgaact atatccgtaa cctggatagt gaaacagggg
     4681 caatggtgcg cctgctggaa gatggcgatt agccatgttt aagggttccg gttccactag
     4741 gtacaattcg atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     4801 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     4861 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     4921 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     4981 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     5041 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     5101 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     5161 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     5221 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     5281 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     5341 tgggccgcct ccccgcatcg ataccgtcga cctcgatcga gacctagaaa aacatggagc
     5401 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
     5461 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
     5521 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     5581 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     5641 tgattggcag aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg
     5701 ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac
     5761 ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga
     5821 gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg
     5881 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     5941 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     6001 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     6061 tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6121 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6181 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6241 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6301 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6361 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6421 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6481 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6541 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6601 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6661 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6721 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6781 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6841 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6901 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6961 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7021 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7081 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7141 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7201 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7261 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7321 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7381 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7441 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7501 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7561 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7621 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7681 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7741 actcatactc ttcctttttc
//
