LOCUS       sequence-360260-        7760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a Arid1a-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-sgArid1a#1/Cre
FEATURES             Location/Qualifiers
     source          1..7760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(132..151)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        323..702
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        704..902
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     857..877
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             920..1100
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1147..1272
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1769..2002
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2187..2231
                     /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    2529..2646
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2728..2976
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2728..2748
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2899..2918
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2998..3073
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3126..3625
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3161..3184)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3558..3577
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3629..3649
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3650..4678
                     /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="SNLLTVHQNLPALPVDATSDEVSKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     misc_feature    4734..5322
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4787..4807)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5205..5216)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5325..5341)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5326..5342)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5851..6031
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6093..6681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6173..6192)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6852..7643,7644..7712))
                     /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     7475..7494
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7713..7760,1..57))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
       61 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
      121 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
      181 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      241 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      301 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      361 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      421 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      481 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      541 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      601 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      661 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      721 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      781 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      841 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      901 agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
      961 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     1021 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     1081 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     1141 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     1201 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     1261 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     1321 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     1381 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     1441 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     1501 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     1561 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccggccgc
     1621 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     1681 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     1741 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     1801 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     1861 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     1921 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     1981 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     2041 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     2101 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     2161 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     2221 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     2281 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     2341 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     2401 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     2461 ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc
     2521 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     2581 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     2641 aattttcggg tttattacag ggacagcaga gatccagttt ggttagtacc gggcccgctc
     2701 tagccctgca ggaactgttt aaactgtgag ggcctatttc ccatgattcc ttcatatttg
     2761 catatacgat acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag
     2821 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2881 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     2941 ttggctttat atatcttgtg gaaaggacca aacaccgatg ggtcccagac tccacaggtt
     3001 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     3061 accgagtcgg tgcttttttg tttggcgcgc ctatggctta attaacatgg cggccgcatt
     3121 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc
     3181 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg
     3241 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct
     3301 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt
     3361 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg
     3421 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg
     3481 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag
     3541 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc
     3601 ctcttcctca tctccgggac ccgggatgcc caagaagaag aggaaggtgt ccaatttact
     3661 gaccgtacac caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttagcaa
     3721 gaacctgatg gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct
     3781 tctgtccgtt tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc
     3841 cgcagaacct gaagatgttc gcgattatct tctatatctt caggcgcgcg gtctggcagt
     3901 aaaaactatc cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc
     3961 acgaccaagt gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt
     4021 tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt
     4081 tcgttcactc atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg
     4141 gattgcttat aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat
     4201 ctcacgtact gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag
     4261 caccgcaggt gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat
     4321 ttccgtctct ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa
     4381 tggtgttgcc gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt
     4441 tgaagcaact catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc
     4501 ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc
     4561 aataccggag atcatgcaag ctggtggctg gaccaatgta aatattgtca tgaactatat
     4621 ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattagcc
     4681 atgtttaagg gttccggttc cactaggtac aattcgatat caagcttatc gataatcaac
     4741 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     4801 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     4861 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     4921 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     4981 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     5041 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     5101 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     5161 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     5221 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     5281 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc
     5341 gatcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct
     5401 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag
     5461 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     5521 aaggggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg
     5581 atctaccaca cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc
     5641 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta
     5701 gaagaagcca atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg
     5761 gatgacccgg agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac
     5821 atggcccgag agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc
     5881 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     5941 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6001 cccttttagt cagtgtggaa aatctctagc agcatgtgag caaaaggcca gcaaaaggcc
     6061 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6121 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6181 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6241 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6301 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6361 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6421 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6481 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6541 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6601 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6661 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6721 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6781 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6841 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6901 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6961 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7021 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7081 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7141 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7201 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7261 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7321 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7381 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7441 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7501 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7561 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7621 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7681 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7741 atttatcagg gttattgtct
//
