LOCUS       sequence-361371-        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#1/Cre
FEATURES             Location/Qualifiers
     source          1..7759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     139..158
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             210..230
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             231..1259
                     /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    1315..1903
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1368..1388)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1786..1797)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1906..1922)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1907..1923)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2432..2612
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2674..3262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2754..2773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3433..4224,4225..4293))
                     /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     4056..4075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4294..4398)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4473..4492)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4664..5043
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5045..5243
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5198..5218
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5261..5441
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5488..5613
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6110..6343
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6528..6572
                     /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    6870..6987
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7069..7317
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7069..7089
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7240..7259
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7338..7413
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        join(7466..7759,1..206)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7501..7524)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg
       61 gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa
      121 ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct
      181 cctcttcctc atctccggga cccgggatgc ccaagaagaa gaggaaggtg tccaatttac
      241 tgaccgtaca ccaaaatttg cctgcattac cggtcgatgc aacgagtgat gaggttcgca
      301 agaacctgat ggacatgttc agggatcgcc aggcgttttc tgagcatacc tggaaaatgc
      361 ttctgtccgt ttgccggtcg tgggcggcat ggtgcaagtt gaataaccgg aaatggtttc
      421 ccgcagaacc tgaagatgtt cgcgattatc ttctatatct tcaggcgcgc ggtctggcag
      481 taaaaactat ccagcaacat ttgggccagc taaacatgct tcatcgtcgg tccgggctgc
      541 cacgaccaag tgacagcaat gctgtttcac tggttatgcg gcggatccga aaagaaaacg
      601 ttgatgccgg tgaacgtgca aaacaggctc tagcgttcga acgcactgat ttcgaccagg
      661 ttcgttcact catggaaaat agcgatcgct gccaggatat acgtaatctg gcatttctgg
      721 ggattgctta taacaccctg ttacgtatag ccgaaattgc caggatcagg gttaaagata
      781 tctcacgtac tgacggtggg agaatgttaa tccatattgg cagaacgaaa acgctggtta
      841 gcaccgcagg tgtagagaag gcacttagcc tgggggtaac taaactggtc gagcgatgga
      901 tttccgtctc tggtgtagct gatgatccga ataactacct gttttgccgg gtcagaaaaa
      961 atggtgttgc cgcgccatct gccaccagcc agctatcaac tcgcgccctg gaagggattt
     1021 ttgaagcaac tcatcgattg atttacggcg ctaaggatga ctctggtcag agatacctgg
     1081 cctggtctgg acacagtgcc cgtgtcggag ccgcgcgaga tatggcccgc gctggagttt
     1141 caataccgga gatcatgcaa gctggtggct ggaccaatgt aaatattgtc atgaactata
     1201 tccgtaacct ggatagtgaa acaggggcaa tggtgcgcct gctggaagat ggcgattagc
     1261 catgtttaag ggttccggtt ccactaggta caattcgata tcaagcttat cgataatcaa
     1321 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1381 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     1441 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     1501 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     1561 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     1621 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     1681 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     1741 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     1801 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     1861 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     1921 cgatcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     1981 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     2041 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     2101 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg
     2161 gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat
     2221 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt
     2281 agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat
     2341 ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca
     2401 catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc
     2461 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     2521 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     2581 acccttttag tcagtgtgga aaatctctag cagcatgtga gcaaaaggcc agcaaaaggc
     2641 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2701 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2761 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2821 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2881 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2941 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3001 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3061 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3121 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3181 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3241 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3301 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3361 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3421 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3481 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3541 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3601 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3661 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3721 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3781 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3841 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3901 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3961 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4021 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4081 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4141 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4201 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4261 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4321 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4381 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     4441 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4501 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     4561 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     4621 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     4681 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     4741 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     4801 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     4861 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     4921 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     4981 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     5041 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     5101 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     5161 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     5221 cggtgggagg tctatataag cagcgcgttt tgcctgtact gggtctctct ggttagacca
     5281 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     5341 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     5401 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
     5461 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
     5521 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
     5581 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
     5641 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
     5701 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
     5761 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
     5821 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
     5881 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
     5941 gcacagcaag cggccggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     6001 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     6061 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     6121 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     6181 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     6241 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     6301 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     6361 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     6421 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     6481 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     6541 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     6601 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     6661 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     6721 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag
     6781 agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt gcgccaattc
     6841 tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac
     6901 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     6961 aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag agatccagtt
     7021 tggttagtac cgggcccgct ctagccctgc aggaactgtt taaactgtga gggcctattt
     7081 cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt
     7141 aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt
     7201 cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta
     7261 acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacc aaacaccgtc
     7321 cgggatcagc taacctggtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
     7381 tatcaacttg aaaaagtggc accgagtcgg tgcttttttg tttggcgcgc ctatggctta
     7441 attaacatgg cggccgcatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga
     7501 gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc
     7561 acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg
     7621 ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc
     7681 aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct
     7741 gagcaatgga agcgggtag
//
