LOCUS       sequence-363743-        7747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a Arid2-targeting gRNA and Cre-recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7747)
  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 7747)
  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-sgArid2#1/Cre
FEATURES             Location/Qualifiers
     source          1..7747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(105..124)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        296..675
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        677..875
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     830..850
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             893..1073
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1120..1245
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1742..1975
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2160..2204
                     /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    2502..2619
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2701..2949
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2701..2721
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2872..2891
                     /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..3585
                     /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     3518..3537
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3589..3609
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3610..4638
                     /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    4694..5282
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4747..4767)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5165..5176)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5285..5301)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5286..5302)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5811..5991
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6053..6641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6133..6152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6812..7603,7604..7672))
                     /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     7435..7454
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7673..7747,1..30))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
       61 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
      121 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      181 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      241 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      301 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      361 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      421 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      481 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      541 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      601 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      661 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      721 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      781 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      841 ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta ctgggtctct
      901 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      961 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     1021 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
     1081 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
     1141 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
     1201 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
     1261 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
     1321 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
     1381 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
     1441 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
     1501 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
     1561 agtaagacca ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata
     1621 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     1681 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     1741 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     1801 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     1861 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     1921 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     1981 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     2041 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     2101 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     2161 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     2221 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     2281 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     2341 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
     2401 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tcggcactgc
     2461 gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
     2521 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     2581 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     2641 agagatccag tttggttagt accgggcccg ctctagccct gcaggaactg tttaaactgt
     2701 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
     2761 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
     2821 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
     2881 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
     2941 ccaaacaccg gcagttccac cacagcagag 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 ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc
     3541 acgtctgccg cgctgttctc ctcttcctca tctccgggac ccgggatgcc caagaagaag
     3601 aggaaggtgt ccaatttact gaccgtacac caaaatttgc ctgcattacc ggtcgatgca
     3661 acgagtgatg aggttcgcaa gaacctgatg gacatgttca gggatcgcca ggcgttttct
     3721 gagcatacct ggaaaatgct tctgtccgtt tgccggtcgt gggcggcatg gtgcaagttg
     3781 aataaccgga aatggtttcc cgcagaacct gaagatgttc gcgattatct tctatatctt
     3841 caggcgcgcg gtctggcagt aaaaactatc cagcaacatt tgggccagct aaacatgctt
     3901 catcgtcggt ccgggctgcc acgaccaagt gacagcaatg ctgtttcact ggttatgcgg
     3961 cggatccgaa aagaaaacgt tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa
     4021 cgcactgatt tcgaccaggt tcgttcactc atggaaaata gcgatcgctg ccaggatata
     4081 cgtaatctgg catttctggg gattgcttat aacaccctgt tacgtatagc cgaaattgcc
     4141 aggatcaggg ttaaagatat ctcacgtact gacggtggga gaatgttaat ccatattggc
     4201 agaacgaaaa cgctggttag caccgcaggt gtagagaagg cacttagcct gggggtaact
     4261 aaactggtcg agcgatggat ttccgtctct ggtgtagctg atgatccgaa taactacctg
     4321 ttttgccggg tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca gctatcaact
     4381 cgcgccctgg aagggatttt tgaagcaact catcgattga tttacggcgc taaggatgac
     4441 tctggtcaga gatacctggc ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat
     4501 atggcccgcg ctggagtttc aataccggag atcatgcaag ctggtggctg gaccaatgta
     4561 aatattgtca tgaactatat ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg
     4621 ctggaagatg gcgattagcc atgtttaagg gttccggttc cactaggtac aattcgatat
     4681 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     4741 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     4801 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     4861 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4921 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     4981 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     5041 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     5101 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     5161 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     5221 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     5281 gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca tggagcaatc acaagtagca
     5341 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
     5401 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     5461 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     5521 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact
     5581 acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac
     5641 cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc
     5701 ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca
     5761 gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct
     5821 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     5881 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     5941 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agcatgtgag
     6001 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6061 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6121 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6181 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6241 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6301 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6361 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6421 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6481 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6541 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6601 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6661 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6721 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6781 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6841 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6901 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6961 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7021 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7081 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7141 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7201 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7261 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7321 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7381 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7441 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7501 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7561 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7621 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7681 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7741 aatgtat
//
