LOCUS       sequence-383729-        7770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing Cre recombinase (driven by mPGK 
            promoter) and an sgRNA against the mouse Nkx2-1 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7770)
  AUTHORS   Lee MC, Cai H, Murray CW, Li C, Shue YT, Andrejka L, He AL, Holzem 
            AME, Drainas AP, Ko JH, Coles GL, Kong C, Zhu S, Zhu C, Wang J, van 
            de Rijn M, Petrov DA, Winslow MM, Sage J
  TITLE     A multiplexed in vivo approach to identify driver genes in small 
            cell lung cancer.
  JOURNAL   Cell Rep. 2023 Jan 13;42(1):111990. doi: 
            10.1016/j.celrep.2023.111990.
  PUBMED    36640300
REFERENCE   2  (bases 1 to 7770)
  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: "Cell Rep.";
            date: "2023-01-13"; volume: "42(1):111990. doi"; pages: " 
            10.1016/j.celrep.2023.111990"
COMMENT     Sequence Label: Lenti-sgNkx2-1#1/Cre
FEATURES             Location/Qualifiers
     source          1..7770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        88..467
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        469..667
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     622..642
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             685..865
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    912..1037
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1534..1767
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1952..1996
                     /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    2294..2411
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2493..2741
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2493..2513
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2664..2683
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        2901..3400
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2936..2959)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3333..3352
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3404..3424
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3425..4453
                     /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    4509..5097
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4562..4582)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4980..4991)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5100..5116)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5101..5117)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5626..5806
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5868..6456)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5948..5967)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6627..7418,7419..7487))
                     /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     7250..7269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7488..7592)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7667..7686)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
       61 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
      121 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      181 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      241 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      301 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      361 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      421 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      481 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      541 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      601 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      661 taagcagcgc gttttgcctg tactgggtct ctctggttag accagatctg agcctgggag
      721 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
      781 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
      841 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
      901 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
      961 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     1021 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     1081 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     1141 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     1201 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     1261 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     1321 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     1381 gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata
     1441 taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag
     1501 agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg
     1561 agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt
     1621 attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca
     1681 tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga
     1741 aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg
     1801 caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa
     1861 tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct taatacactc
     1921 cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga
     1981 taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt
     2041 attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat
     2101 agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc
     2161 gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag
     2221 atccattcga ttagtgaacg gatcggcact gcgtgcgcca attctgcaga caaatggcag
     2281 tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     2341 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     2401 ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttggtta gtaccgggcc
     2461 cgctctagcc ctgcaggaac tgtttaaact gtgagggcct atttcccatg attccttcat
     2521 atttgcatat acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca
     2581 caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag
     2641 ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg
     2701 atttcttggc tttatatatc ttgtggaaag gaccaaacac cgcaactgca gcaggacagc
     2761 gggtttaaga gctatgctgg aaacagcata gcaagtttaa ataaggctag tccgttatca
     2821 acttgaaaaa gtggcaccga gtcggtgctt ttttgtttgg cgcgcctatg gcttaattaa
     2881 catggcggcc gcattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg
     2941 cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca
     3001 ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc
     3061 ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac
     3121 gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca
     3181 atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg
     3241 ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg
     3301 ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc
     3361 tgccgcgctg ttctcctctt cctcatctcc gggacccggg atgcccaaga agaagaggaa
     3421 ggtgtccaat ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg atgcaacgag
     3481 tgatgaggtt cgcaagaacc tgatggacat gttcagggat cgccaggcgt tttctgagca
     3541 tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca agttgaataa
     3601 ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat tatcttctat atcttcaggc
     3661 gcgcggtctg gcagtaaaaa ctatccagca acatttgggc cagctaaaca tgcttcatcg
     3721 tcggtccggg ctgccacgac caagtgacag caatgctgtt tcactggtta tgcggcggat
     3781 ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag gctctagcgt tcgaacgcac
     3841 tgatttcgac caggttcgtt cactcatgga aaatagcgat cgctgccagg atatacgtaa
     3901 tctggcattt ctggggattg cttataacac cctgttacgt atagccgaaa ttgccaggat
     3961 cagggttaaa gatatctcac gtactgacgg tgggagaatg ttaatccata ttggcagaac
     4021 gaaaacgctg gttagcaccg caggtgtaga gaaggcactt agcctggggg taactaaact
     4081 ggtcgagcga tggatttccg tctctggtgt agctgatgat ccgaataact acctgttttg
     4141 ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc agccagctat caactcgcgc
     4201 cctggaaggg atttttgaag caactcatcg attgatttac ggcgctaagg atgactctgg
     4261 tcagagatac ctggcctggt ctggacacag tgcccgtgtc ggagccgcgc gagatatggc
     4321 ccgcgctgga gtttcaatac cggagatcat gcaagctggt ggctggacca atgtaaatat
     4381 tgtcatgaac tatatccgta acctggatag tgaaacaggg gcaatggtgc gcctgctgga
     4441 agatggcgat tagccatgtt taagggttcc ggttccacta ggtacaattc gatatcaagc
     4501 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     4561 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     4621 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     4681 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     4741 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     4801 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     4861 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     4921 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     4981 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     5041 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     5101 gataccgtcg acctcgatcg agacctagaa aaacatggag caatcacaag tagcaataca
     5161 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt
     5221 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     5281 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     5341 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca gaactacaca
     5401 ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct agtaccagtt
     5461 gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt acaccctgtg
     5521 agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt tgacagccgc
     5581 ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc tctctggtta
     5641 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     5701 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     5761 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagcat gtgagcaaaa
     5821 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5881 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5941 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6001 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6061 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6121 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6181 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6241 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6301 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6361 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6421 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6481 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6541 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6601 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6661 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6721 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6781 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6841 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6901 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6961 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7021 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7081 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7141 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7201 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7261 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7321 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7381 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7441 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7501 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7561 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7621 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     7681 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     7741 gcgcgagcaa aatttaagct acaacaaggc
//
