LOCUS       sequence-352719-        8250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses  Cebpa_v1 (mU6), Cebpd_v1 (hU6) gRNAs and Cre-recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8250)
  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 8250)
  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_v1-hu6-sgCebpd_v1
FEATURES             Location/Qualifiers
     source          1..8250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             209..389
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(451..1039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(531..550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1210..2001,2002..2070))
                     /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     1833..1852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2071..2175)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2250..2269)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2441..2820
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2822..3020
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2975..2995
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3038..3218
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3265..3390
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3887..4120
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4305..4349
                     /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    4647..4764
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     5077..5097
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     5323..5343
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5494..5513
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        5734..6233
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5769..5792)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6166..6185
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6237..6257
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6258..7286
                     /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    7342..7930
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7395..7415)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7813..7824)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7933..7949)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7934..7950)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
       61 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
      121 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
      181 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
      241 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
      301 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
      361 cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag
      421 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      481 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      541 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      601 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      661 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      721 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      781 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      841 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
      901 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      961 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1021 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1081 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1141 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1201 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1261 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     1321 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     1381 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     1441 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     1501 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     1561 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     1621 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     1681 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     1741 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     1801 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     1861 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     1921 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     1981 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2041 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2101 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2161 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     2221 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2281 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     2341 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     2401 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
     2461 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     2521 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     2581 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     2641 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     2701 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     2761 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     2821 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     2881 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     2941 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     3001 tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat
     3061 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     3121 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     3181 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     3241 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     3301 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     3361 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     3421 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     3481 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     3541 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     3601 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     3661 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     3721 cagcaagcgg ccggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     3781 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     3841 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     3901 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     3961 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     4021 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     4081 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     4141 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     4201 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     4261 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     4321 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     4381 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     4441 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     4501 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     4561 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     4621 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     4681 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     4741 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     4801 ttagtaccgg gcccgctcta gccctgcagg aactgtttaa actgtgatcc gacgcgccat
     4861 ctctaggccc gcgccggccc cctcgcacgg acttgtggga gaagctcggc tactcccctg
     4921 ccccggttaa tttgcatata atatttccta gtaactatag aggcttaatg tgcgataaaa
     4981 gacagataat ctgttctttt taatactagc tacattttac atgataggct tggatttcta
     5041 taagagatac aaatactaaa ttattatttt aaaaaacagc acaaaaggaa actcacccta
     5101 actgtaaagt aattgtgtgt tttgagacta taagtatccc ttggagaacc accttgttgg
     5161 tgctcgcaga tgccgcccag gtttgagagc taagcagaaa gctgcatagc aagttcaaat
     5221 aaggctagtc cgtacacaac ttgaaaaagt ggcagccgag tcggctgctt tttttgctcg
     5281 aatctacact cagctatggc gcgccccaag gtcgggcagg aagagggcct atttcccatg
     5341 attccttcat atttgcatat acgatacaag gctgttagag agataattgg aattaatttg
     5401 actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg
     5461 tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga
     5521 aagtatttcg atttcttggc tttatatatc ttgtggaaag ccagaaacat ggctcaagcg
     5581 cgaacccgac tggtttcaga gctaagcaca agagtgcata gcaagttgaa ataaggctag
     5641 tccgtttaca acttgaaaaa gtggcacccg agtcgggtgc tttttttgtt tggcgcgcct
     5701 atggcttaat taacatggcg gccgcattct accgggtagg ggaggcgctt ttcccaaggc
     5761 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
     5821 ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc
     5881 ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg
     5941 cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca
     6001 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct
     6061 ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg
     6121 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
     6181 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggaccc gggatgccca
     6241 agaagaagag gaaggtgtcc aatttactga ccgtacacca aaatttgcct gcattaccgg
     6301 tcgatgcaac gagtgatgag gttcgcaaga acctgatgga catgttcagg gatcgccagg
     6361 cgttttctga gcatacctgg aaaatgcttc tgtccgtttg ccggtcgtgg gcggcatggt
     6421 gcaagttgaa taaccggaaa tggtttcccg cagaacctga agatgttcgc gattatcttc
     6481 tatatcttca ggcgcgcggt ctggcagtaa aaactatcca gcaacatttg ggccagctaa
     6541 acatgcttca tcgtcggtcc gggctgccac gaccaagtga cagcaatgct gtttcactgg
     6601 ttatgcggcg gatccgaaaa gaaaacgttg atgccggtga acgtgcaaaa caggctctag
     6661 cgttcgaacg cactgatttc gaccaggttc gttcactcat ggaaaatagc gatcgctgcc
     6721 aggatatacg taatctggca tttctgggga ttgcttataa caccctgtta cgtatagccg
     6781 aaattgccag gatcagggtt aaagatatct cacgtactga cggtgggaga atgttaatcc
     6841 atattggcag aacgaaaacg ctggttagca ccgcaggtgt agagaaggca cttagcctgg
     6901 gggtaactaa actggtcgag cgatggattt ccgtctctgg tgtagctgat gatccgaata
     6961 actacctgtt ttgccgggtc agaaaaaatg gtgttgccgc gccatctgcc accagccagc
     7021 tatcaactcg cgccctggaa gggatttttg aagcaactca tcgattgatt tacggcgcta
     7081 aggatgactc tggtcagaga tacctggcct ggtctggaca cagtgcccgt gtcggagccg
     7141 cgcgagatat ggcccgcgct ggagtttcaa taccggagat catgcaagct ggtggctgga
     7201 ccaatgtaaa tattgtcatg aactatatcc gtaacctgga tagtgaaaca ggggcaatgg
     7261 tgcgcctgct ggaagatggc gattagccat gtttaagggt tccggttcca ctaggtacaa
     7321 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     7381 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     7441 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     7501 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     7561 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     7621 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     7681 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     7741 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     7801 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     7861 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     7921 gcctccccgc atcgataccg tcgacctcga tcgagaccta gaaaaacatg gagcaatcac
     7981 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
     8041 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     8101 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     8161 acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg
     8221 gcagaactac acaccagggc cagggatcag
//
