LOCUS       sequence-361372-        7759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a Atf7ip-targeting gRNA and Cre-recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7759)
  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 7759)
  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-sgAtf7ip#2/Cre
FEATURES             Location/Qualifiers
     source          1..7759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(89..108)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        280..659
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        661..859
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     814..834
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             877..1057
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1104..1229
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1726..1959
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2144..2188
                     /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    2486..2603
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2685..2933
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2685..2705
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2856..2875
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2954..3029
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3082..3581
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3117..3140)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3514..3533
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3585..3605
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3606..4634
                     /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    4690..5278
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4743..4763)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5161..5172)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5281..5297)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5282..5298)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5807..5987
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6049..6637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6129..6148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6808..7599,7600..7668))
                     /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     7431..7450
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7669..7759,1..14))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
       61 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
      121 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
      181 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
      241 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
      301 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      361 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      421 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      481 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      541 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      601 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      661 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      721 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      781 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      841 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      901 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      961 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     1021 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
     1081 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
     1141 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
     1201 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
     1261 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
     1321 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
     1381 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     1441 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     1501 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     1561 agcaagcggc cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
     1621 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     1681 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     1741 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     1801 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     1861 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     1921 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     1981 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     2041 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     2101 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     2161 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     2221 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     2281 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     2341 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag
     2401 agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc caattctgca
     2461 gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg
     2521 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     2581 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttggt
     2641 tagtaccggg cccgctctag ccctgcagga actgtttaaa ctgtgagggc ctatttccca
     2701 tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt
     2761 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
     2821 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     2881 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggaccaaac accgaacagt
     2941 aatgtcgtcg agagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
     3001 aacttgaaaa agtggcaccg agtcggtgct tttttgtttg gcgcgcctat ggcttaatta
     3061 acatggcggc cgcattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
     3121 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
     3181 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
     3241 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
     3301 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
     3361 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
     3421 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
     3481 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
     3541 ctgccgcgct gttctcctct tcctcatctc cgggacccgg gatgcccaag aagaagagga
     3601 aggtgtccaa tttactgacc gtacaccaaa atttgcctgc attaccggtc gatgcaacga
     3661 gtgatgaggt tcgcaagaac ctgatggaca tgttcaggga tcgccaggcg ttttctgagc
     3721 atacctggaa aatgcttctg tccgtttgcc ggtcgtgggc ggcatggtgc aagttgaata
     3781 accggaaatg gtttcccgca gaacctgaag atgttcgcga ttatcttcta tatcttcagg
     3841 cgcgcggtct ggcagtaaaa actatccagc aacatttggg ccagctaaac atgcttcatc
     3901 gtcggtccgg gctgccacga ccaagtgaca gcaatgctgt ttcactggtt atgcggcgga
     3961 tccgaaaaga aaacgttgat gccggtgaac gtgcaaaaca ggctctagcg ttcgaacgca
     4021 ctgatttcga ccaggttcgt tcactcatgg aaaatagcga tcgctgccag gatatacgta
     4081 atctggcatt tctggggatt gcttataaca ccctgttacg tatagccgaa attgccagga
     4141 tcagggttaa agatatctca cgtactgacg gtgggagaat gttaatccat attggcagaa
     4201 cgaaaacgct ggttagcacc gcaggtgtag agaaggcact tagcctgggg gtaactaaac
     4261 tggtcgagcg atggatttcc gtctctggtg tagctgatga tccgaataac tacctgtttt
     4321 gccgggtcag aaaaaatggt gttgccgcgc catctgccac cagccagcta tcaactcgcg
     4381 ccctggaagg gatttttgaa gcaactcatc gattgattta cggcgctaag gatgactctg
     4441 gtcagagata cctggcctgg tctggacaca gtgcccgtgt cggagccgcg cgagatatgg
     4501 cccgcgctgg agtttcaata ccggagatca tgcaagctgg tggctggacc aatgtaaata
     4561 ttgtcatgaa ctatatccgt aacctggata gtgaaacagg ggcaatggtg cgcctgctgg
     4621 aagatggcga ttagccatgt ttaagggttc cggttccact aggtacaatt cgatatcaag
     4681 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     4741 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     4801 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     4861 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     4921 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     4981 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     5041 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     5101 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     5161 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     5221 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     5281 cgataccgtc gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     5341 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt
     5401 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     5461 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     5521 tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc agaactacac
     5581 accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     5641 tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt
     5701 gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg
     5761 cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt
     5821 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     5881 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     5941 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagca tgtgagcaaa
     6001 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6061 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6121 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6181 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6241 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6301 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6361 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6421 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6481 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6541 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6601 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6661 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6721 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6781 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6841 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6901 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6961 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7021 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7081 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7141 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7201 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7261 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7321 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7381 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7441 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7501 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7561 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7621 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7681 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7741 tatttagaaa aataaacaa
//
