LOCUS       sequence-363745-        7760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a Arid1b-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-sgArid1b#1/Cre
FEATURES             Location/Qualifiers
     source          1..7760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(104..123)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        295..674
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        676..874
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     829..849
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             892..1072
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1119..1244
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1741..1974
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2159..2203
                     /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    2501..2618
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2700..2948
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2700..2720
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2871..2890
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2970..3045
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3098..3597
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3133..3156)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3530..3549
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3601..3621
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3622..4650
                     /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    4706..5294
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4759..4779)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5177..5188)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5297..5313)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5298..5314)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5823..6003
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6065..6653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6145..6164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6824..7615,7616..7684))
                     /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     7447..7466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7685..7760,1..29))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
       61 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
      121 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      181 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      241 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      301 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      361 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      421 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      481 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      541 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      601 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      661 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      721 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      781 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      841 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
      901 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      961 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     1021 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     1081 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     1141 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     1201 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     1261 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     1321 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     1381 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     1441 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     1501 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     1561 gtaagaccac cgcacagcaa gcggccggcc gctgatcttc agacctggag gaggagatat
     1621 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     1681 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     1741 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     1801 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     1861 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     1921 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     1981 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     2041 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     2101 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     2161 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     2221 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     2281 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     2341 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     2401 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg
     2461 tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     2521 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     2581 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     2641 gagatccagt ttggttagta ccgggcccgc tctagccctg caggaactgt ttaaactgtg
     2701 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
     2761 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
     2821 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
     2881 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     2941 caaacaccgc taacgttcat ggaagagagg ttttagagct agaaatagca agttaaaata
     3001 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgtttggcgc
     3061 gcctatggct taattaacat ggcggccgca ttctaccggg taggggaggc gcttttccca
     3121 aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc
     3181 ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt
     3241 ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag
     3301 ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg
     3361 gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt
     3421 tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg
     3481 ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg
     3541 cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg acccgggatg
     3601 cccaagaaga agaggaaggt gtccaattta ctgaccgtac accaaaattt gcctgcatta
     3661 ccggtcgatg caacgagtga tgaggttcgc aagaacctga tggacatgtt cagggatcgc
     3721 caggcgtttt ctgagcatac ctggaaaatg cttctgtccg tttgccggtc gtgggcggca
     3781 tggtgcaagt tgaataaccg gaaatggttt cccgcagaac ctgaagatgt tcgcgattat
     3841 cttctatatc ttcaggcgcg cggtctggca gtaaaaacta tccagcaaca tttgggccag
     3901 ctaaacatgc ttcatcgtcg gtccgggctg ccacgaccaa gtgacagcaa tgctgtttca
     3961 ctggttatgc ggcggatccg aaaagaaaac gttgatgccg gtgaacgtgc aaaacaggct
     4021 ctagcgttcg aacgcactga tttcgaccag gttcgttcac tcatggaaaa tagcgatcgc
     4081 tgccaggata tacgtaatct ggcatttctg gggattgctt ataacaccct gttacgtata
     4141 gccgaaattg ccaggatcag ggttaaagat atctcacgta ctgacggtgg gagaatgtta
     4201 atccatattg gcagaacgaa aacgctggtt agcaccgcag gtgtagagaa ggcacttagc
     4261 ctgggggtaa ctaaactggt cgagcgatgg atttccgtct ctggtgtagc tgatgatccg
     4321 aataactacc tgttttgccg ggtcagaaaa aatggtgttg ccgcgccatc tgccaccagc
     4381 cagctatcaa ctcgcgccct ggaagggatt tttgaagcaa ctcatcgatt gatttacggc
     4441 gctaaggatg actctggtca gagatacctg gcctggtctg gacacagtgc ccgtgtcgga
     4501 gccgcgcgag atatggcccg cgctggagtt tcaataccgg agatcatgca agctggtggc
     4561 tggaccaatg taaatattgt catgaactat atccgtaacc tggatagtga aacaggggca
     4621 atggtgcgcc tgctggaaga tggcgattag ccatgtttaa gggttccggt tccactaggt
     4681 acaattcgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     4741 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     4801 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     4861 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     4921 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     4981 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5041 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     5101 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     5161 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     5221 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     5281 ggccgcctcc ccgcatcgat accgtcgacc tcgatcgaga cctagaaaaa catggagcaa
     5341 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
     5401 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     5461 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     5521 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     5581 attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct
     5641 acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga gagaacaccc
     5701 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtattagagt
     5761 ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat ccggactgta
     5821 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     5881 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     5941 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6001 gcagcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6061 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6121 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6181 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6241 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6301 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6361 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6421 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6481 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6541 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6601 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6661 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6721 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6781 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6841 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6901 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6961 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7021 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7081 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7141 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7201 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7261 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7321 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7381 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7441 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7501 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7561 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7621 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7681 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7741 gatacatatt tgaatgtatt
//
