LOCUS       sequence-361385-        7760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a Bap1-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-sgBap1#1/Cre
FEATURES             Location/Qualifiers
     source          1..7760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(112..131)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        303..682
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        684..882
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     837..857
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             900..1080
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1127..1252
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1749..1982
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2167..2211
                     /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    2509..2626
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2708..2956
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2708..2728
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2879..2898
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2978..3053
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3106..3605
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3141..3164)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3538..3557
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3609..3629
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3630..4658
                     /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    4714..5302
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4767..4787)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5185..5196)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5305..5321)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5306..5322)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5831..6011
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6073..6661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6153..6172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6832..7623,7624..7692))
                     /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     7455..7474
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7693..7760,1..37))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
       61 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
      121 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
      181 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
      241 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
      301 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
      361 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      421 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      481 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      541 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      601 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      661 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      721 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      781 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      841 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtactg
      901 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
      961 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     1021 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     1081 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     1141 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     1201 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     1261 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     1321 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     1381 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     1441 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     1501 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     1561 aaacaaaagt aagaccaccg cacagcaagc ggccggccgc tgatcttcag acctggagga
     1621 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     1681 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     1741 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     1801 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     1861 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     1921 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     1981 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     2041 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     2101 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     2161 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     2221 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     2281 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     2341 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     2401 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg
     2461 gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa
     2521 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     2581 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     2641 ggacagcaga gatccagttt ggttagtacc gggcccgctc tagccctgca ggaactgttt
     2701 aaactgtgag ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt
     2761 tagagagata attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg
     2821 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga
     2881 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg
     2941 gaaaggacca aacaccgcca acgtagaaac cttgcgggtt ttagagctag aaatagcaag
     3001 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg
     3061 tttggcgcgc ctatggctta attaacatgg cggccgcatt ctaccgggta ggggaggcgc
     3121 ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca
     3181 caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt
     3241 tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg
     3301 ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg
     3361 tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat
     3421 agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg
     3481 ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat
     3541 tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggac
     3601 ccgggatgcc caagaagaag aggaaggtgt ccaatttact gaccgtacac caaaatttgc
     3661 ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa gaacctgatg gacatgttca
     3721 gggatcgcca ggcgttttct gagcatacct ggaaaatgct tctgtccgtt tgccggtcgt
     3781 gggcggcatg gtgcaagttg aataaccgga aatggtttcc cgcagaacct gaagatgttc
     3841 gcgattatct tctatatctt caggcgcgcg gtctggcagt aaaaactatc cagcaacatt
     3901 tgggccagct aaacatgctt catcgtcggt ccgggctgcc acgaccaagt gacagcaatg
     3961 ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt tgatgccggt gaacgtgcaa
     4021 aacaggctct agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc atggaaaata
     4081 gcgatcgctg ccaggatata cgtaatctgg catttctggg gattgcttat aacaccctgt
     4141 tacgtatagc cgaaattgcc aggatcaggg ttaaagatat ctcacgtact gacggtggga
     4201 gaatgttaat ccatattggc agaacgaaaa cgctggttag caccgcaggt gtagagaagg
     4261 cacttagcct gggggtaact aaactggtcg agcgatggat ttccgtctct ggtgtagctg
     4321 atgatccgaa taactacctg ttttgccggg tcagaaaaaa tggtgttgcc gcgccatctg
     4381 ccaccagcca gctatcaact cgcgccctgg aagggatttt tgaagcaact catcgattga
     4441 tttacggcgc taaggatgac tctggtcaga gatacctggc ctggtctgga cacagtgccc
     4501 gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc aataccggag atcatgcaag
     4561 ctggtggctg gaccaatgta aatattgtca tgaactatat ccgtaacctg gatagtgaaa
     4621 caggggcaat ggtgcgcctg ctggaagatg gcgattagcc atgtttaagg gttccggttc
     4681 cactaggtac aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
     4741 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     4801 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     4861 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     4921 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4981 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     5041 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     5101 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     5161 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     5221 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     5281 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca
     5341 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
     5401 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
     5461 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     5521 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
     5581 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
     5641 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
     5701 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
     5761 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
     5821 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     5881 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     5941 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     6001 aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6061 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6121 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6181 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6241 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6301 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6361 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6421 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6481 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6541 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6601 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6661 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6721 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6781 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6841 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6901 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6961 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7021 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7081 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7141 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7201 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7261 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7321 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7381 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7441 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7501 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7561 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7621 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7681 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7741 catgagcgga tacatatttg
//
