LOCUS       sequence-363646-        7760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a Gata3-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-sgGata3#2/Cre
FEATURES             Location/Qualifiers
     source          1..7760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(93..112)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        284..663
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        665..863
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     818..838
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             881..1061
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1108..1233
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1730..1963
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2148..2192
                     /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    2490..2607
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2689..2937
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2689..2709
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2860..2879
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2959..3034
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3087..3586
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3122..3145)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3519..3538
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3590..3610
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3611..4639
                     /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    4695..5283
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4748..4768)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5166..5177)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5286..5302)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5287..5303)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5812..5992
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6054..6642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6134..6153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6813..7604,7605..7673))
                     /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     7436..7455
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7674..7760,1..18))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
       61 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
      121 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
      181 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
      241 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
      301 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      361 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      421 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      481 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      541 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      601 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      661 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      721 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      781 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      841 cggtgggagg tctatataag cagcgcgttt tgcctgtact gggtctctct ggttagacca
      901 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
      961 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     1021 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
     1081 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
     1141 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
     1201 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
     1261 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
     1321 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
     1381 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
     1441 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
     1501 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
     1561 gcacagcaag cggccggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     1621 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     1681 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     1741 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     1801 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     1861 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     1921 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     1981 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     2041 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     2101 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     2161 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     2221 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     2281 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     2341 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag
     2401 agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt gcgccaattc
     2461 tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac
     2521 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     2581 aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag agatccagtt
     2641 tggttagtac cgggcccgct ctagccctgc aggaactgtt taaactgtga gggcctattt
     2701 cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt
     2761 aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt
     2821 cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta
     2881 acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacc aaacaccgag
     2941 gtatcctccg acccaccagt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
     3001 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt gtttggcgcg cctatggctt
     3061 aattaacatg gcggccgcat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
     3121 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
     3181 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
     3241 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
     3301 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
     3361 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
     3421 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
     3481 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
     3541 cacgtctgcc gcgctgttct cctcttcctc atctccggga cccgggatgc ccaagaagaa
     3601 gaggaaggtg tccaatttac tgaccgtaca ccaaaatttg cctgcattac cggtcgatgc
     3661 aacgagtgat gaggttcgca agaacctgat ggacatgttc agggatcgcc aggcgttttc
     3721 tgagcatacc tggaaaatgc ttctgtccgt ttgccggtcg tgggcggcat ggtgcaagtt
     3781 gaataaccgg aaatggtttc ccgcagaacc tgaagatgtt cgcgattatc ttctatatct
     3841 tcaggcgcgc ggtctggcag taaaaactat ccagcaacat ttgggccagc taaacatgct
     3901 tcatcgtcgg tccgggctgc cacgaccaag tgacagcaat gctgtttcac tggttatgcg
     3961 gcggatccga aaagaaaacg ttgatgccgg tgaacgtgca aaacaggctc tagcgttcga
     4021 acgcactgat ttcgaccagg ttcgttcact catggaaaat agcgatcgct gccaggatat
     4081 acgtaatctg gcatttctgg ggattgctta taacaccctg ttacgtatag ccgaaattgc
     4141 caggatcagg gttaaagata tctcacgtac tgacggtggg agaatgttaa tccatattgg
     4201 cagaacgaaa acgctggtta gcaccgcagg tgtagagaag gcacttagcc tgggggtaac
     4261 taaactggtc gagcgatgga tttccgtctc tggtgtagct gatgatccga ataactacct
     4321 gttttgccgg gtcagaaaaa atggtgttgc cgcgccatct gccaccagcc agctatcaac
     4381 tcgcgccctg gaagggattt ttgaagcaac tcatcgattg atttacggcg ctaaggatga
     4441 ctctggtcag agatacctgg cctggtctgg acacagtgcc cgtgtcggag ccgcgcgaga
     4501 tatggcccgc gctggagttt caataccgga gatcatgcaa gctggtggct ggaccaatgt
     4561 aaatattgtc atgaactata tccgtaacct ggatagtgaa acaggggcaa tggtgcgcct
     4621 gctggaagat ggcgattagc catgtttaag ggttccggtt ccactaggta caattcgata
     4681 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     4741 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     4801 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     4861 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     4921 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     4981 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     5041 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     5101 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     5161 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     5221 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     5281 cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat cacaagtagc
     5341 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg
     5401 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat
     5461 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     5521 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac
     5581 tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta
     5641 ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac
     5701 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac
     5761 agccgcctag catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc
     5821 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     5881 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     5941 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagcatgtga
     6001 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6061 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6121 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6181 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6241 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6301 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6361 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6421 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6481 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6541 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6601 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6661 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6721 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6781 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6841 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6901 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6961 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7021 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7081 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7141 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7201 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7261 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7321 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7381 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7441 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7501 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7561 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7621 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7681 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7741 gaatgtattt agaaaaataa
//
