LOCUS       sequence-351954-        8249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses  Cebpa_v2 (mU6), Cebpb_v2(hU6) gRNAs and Cre-recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8249)
  AUTHORS   Murray CW, Brady JJ, Han M, Cai H, Tsai MK, Pierce SE, Cheng R, 
            Demeter J, Feldser DM, Jackson PK, Shackelford DB, Winslow MM
  TITLE     LKB1 drives stasis and C/EBP-mediated reprogramming to an alveolar 
            type II fate in lung cancer.
  JOURNAL   Nat Commun. 2022 Feb 28;13(1):1090. doi: 10.1038/s41467-022-28619-8.
  PUBMED    35228570
REFERENCE   2  (bases 1 to 8249)
  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: "Nat 
            Commun."; date: "2022-02-28"; volume: "13(1):1090. doi"; pages: " 
            10.1038/s41467-022-28619-8"
COMMENT     Sequence Label: mU6-sgCebpa_v2-hu6-sgCebpb_v2
FEATURES             Location/Qualifiers
     source          1..8249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     184..203
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             255..275
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             276..1304
                     /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    1360..1948
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1413..1433)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1831..1842)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1951..1967)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1952..1968)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2477..2657
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2719..3307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2799..2818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3478..4269,4270..4338))
                     /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     4101..4120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4339..4443)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4518..4537)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4709..5088
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5090..5288
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5243..5263
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5306..5486
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5533..5658
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6155..6388
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6573..6617
                     /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    6915..7032
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     7345..7365
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     7591..7611
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7762..7781
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        join(8001..8249,1..251)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8036..8059)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg
       61 gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg
      121 ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc
      181 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc
      241 cgggacccgg gatgcccaag aagaagagga aggtgtccaa tttactgacc gtacaccaaa
      301 atttgcctgc attaccggtc gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca
      361 tgttcaggga tcgccaggcg ttttctgagc atacctggaa aatgcttctg tccgtttgcc
      421 ggtcgtgggc ggcatggtgc aagttgaata accggaaatg gtttcccgca gaacctgaag
      481 atgttcgcga ttatcttcta tatcttcagg cgcgcggtct ggcagtaaaa actatccagc
      541 aacatttggg ccagctaaac atgcttcatc gtcggtccgg gctgccacga ccaagtgaca
      601 gcaatgctgt ttcactggtt atgcggcgga tccgaaaaga aaacgttgat gccggtgaac
      661 gtgcaaaaca ggctctagcg ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg
      721 aaaatagcga tcgctgccag gatatacgta atctggcatt tctggggatt gcttataaca
      781 ccctgttacg tatagccgaa attgccagga tcagggttaa agatatctca cgtactgacg
      841 gtgggagaat gttaatccat attggcagaa cgaaaacgct ggttagcacc gcaggtgtag
      901 agaaggcact tagcctgggg gtaactaaac tggtcgagcg atggatttcc gtctctggtg
      961 tagctgatga tccgaataac tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc
     1021 catctgccac cagccagcta tcaactcgcg ccctggaagg gatttttgaa gcaactcatc
     1081 gattgattta cggcgctaag gatgactctg gtcagagata cctggcctgg tctggacaca
     1141 gtgcccgtgt cggagccgcg cgagatatgg cccgcgctgg agtttcaata ccggagatca
     1201 tgcaagctgg tggctggacc aatgtaaata ttgtcatgaa ctatatccgt aacctggata
     1261 gtgaaacagg ggcaatggtg cgcctgctgg aagatggcga ttagccatgt ttaagggttc
     1321 cggttccact aggtacaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
     1381 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     1441 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     1501 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     1561 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     1621 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     1681 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     1741 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     1801 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     1861 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     1921 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgatc gagacctaga
     1981 aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct
     2041 agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc
     2101 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     2161 agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca
     2221 aggctacttc cctgattggc agaactacac accagggcca gggatcagat atccactgac
     2281 ctttggatgg tgctacaagc tagtaccagt tgagcaagag aaggtagaag aagccaatga
     2341 aggagagaac acccgcttgt tacaccctgt gagcctgcat gggatggatg acccggagag
     2401 agaagtatta gagtggaggt ttgacagccg cctagcattt catcacatgg cccgagagct
     2461 gcatccggac tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     2521 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     2581 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     2641 gtggaaaatc tctagcagca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2701 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2761 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2821 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2881 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2941 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3001 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3061 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3121 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3181 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3241 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3301 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3361 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3421 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3481 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3541 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3601 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3661 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3721 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3781 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3841 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3901 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3961 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4021 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4081 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4141 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4201 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4261 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4321 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4381 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4441 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     4501 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     4561 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     4621 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     4681 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     4741 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     4801 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     4861 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     4921 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     4981 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     5041 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     5101 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     5161 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     5221 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     5281 ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga
     5341 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     5401 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     5461 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
     5521 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
     5581 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
     5641 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
     5701 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
     5761 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     5821 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     5881 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     5941 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     6001 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     6061 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     6121 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     6181 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     6241 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     6301 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     6361 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     6421 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     6481 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     6541 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     6601 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     6661 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     6721 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     6781 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     6841 gatccattcg attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca
     6901 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     6961 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     7021 attcaaaatt ttcgggttta ttacagggac agcagagatc cagtttggtt agtaccgggc
     7081 ccgctctagc cctgcaggaa ctgtttaaac tgtgatccga cgcgccatct ctaggcccgc
     7141 gccggccccc tcgcacggac ttgtgggaga agctcggcta ctcccctgcc ccggttaatt
     7201 tgcatataat atttcctagt aactatagag gcttaatgtg cgataaaaga cagataatct
     7261 gttcttttta atactagcta cattttacat gataggcttg gatttctata agagatacaa
     7321 atactaaatt attattttaa aaaacagcac aaaaggaaac tcaccctaac tgtaaagtaa
     7381 ttgtgtgttt tgagactata agtatccctt ggagaaccac cttgttggca tcagcgccta
     7441 catcgaccgt ttgagagcta agcagaaagc tgcatagcaa gttcaaataa ggctagtccg
     7501 tacacaactt gaaaaagtgg cagccgagtc ggctgctttt tttgctcgaa tctacactca
     7561 gctatggcgc gccccaaggt cgggcaggaa gagggcctat ttcccatgat tccttcatat
     7621 ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca
     7681 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     7741 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     7801 ttcttggctt tatatatctt gtggaaagcc agaaacatgg tgctcgccaa tggccggctg
     7861 tttcagagct aagcacaaga gtgcatagca agttgaaata aggctagtcc gtttacaact
     7921 tgaaaaagtg gcacccgagt cgggtgcttt ttttgtttgg cgcgcctatg gcttaattaa
     7981 catggcggcc gcattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg
     8041 cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca
     8101 ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc
     8161 ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac
     8221 gtgacaaatg gaagtagcac gtctcacta
//
