LOCUS       sequence-351920-        8250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses  Cebpb_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-sgCebpb_v1-hu6-sgCebpd_v1
FEATURES             Location/Qualifiers
     source          1..8250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     45..64
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             116..136
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             137..1165
                     /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    1221..1809
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1274..1294)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1692..1703)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1812..1828)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1813..1829)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2338..2518
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2580..3168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2660..2679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3339..4130,4131..4199))
                     /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     3962..3981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4200..4304)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4379..4398)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4570..4949
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4951..5149
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5104..5124
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5167..5347
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5394..5519
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6016..6249
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6434..6478
                     /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    6776..6893
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     7206..7226
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     7452..7472
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7623..7642
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        join(7863..8250,1..112)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7898..7921)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
       61 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggacccg ggatgcccaa
      121 gaagaagagg aaggtgtcca atttactgac cgtacaccaa aatttgcctg cattaccggt
      181 cgatgcaacg agtgatgagg ttcgcaagaa cctgatggac atgttcaggg atcgccaggc
      241 gttttctgag catacctgga aaatgcttct gtccgtttgc cggtcgtggg cggcatggtg
      301 caagttgaat aaccggaaat ggtttcccgc agaacctgaa gatgttcgcg attatcttct
      361 atatcttcag gcgcgcggtc tggcagtaaa aactatccag caacatttgg gccagctaaa
      421 catgcttcat cgtcggtccg ggctgccacg accaagtgac agcaatgctg tttcactggt
      481 tatgcggcgg atccgaaaag aaaacgttga tgccggtgaa cgtgcaaaac aggctctagc
      541 gttcgaacgc actgatttcg accaggttcg ttcactcatg gaaaatagcg atcgctgcca
      601 ggatatacgt aatctggcat ttctggggat tgcttataac accctgttac gtatagccga
      661 aattgccagg atcagggtta aagatatctc acgtactgac ggtgggagaa tgttaatcca
      721 tattggcaga acgaaaacgc tggttagcac cgcaggtgta gagaaggcac ttagcctggg
      781 ggtaactaaa ctggtcgagc gatggatttc cgtctctggt gtagctgatg atccgaataa
      841 ctacctgttt tgccgggtca gaaaaaatgg tgttgccgcg ccatctgcca ccagccagct
      901 atcaactcgc gccctggaag ggatttttga agcaactcat cgattgattt acggcgctaa
      961 ggatgactct ggtcagagat acctggcctg gtctggacac agtgcccgtg tcggagccgc
     1021 gcgagatatg gcccgcgctg gagtttcaat accggagatc atgcaagctg gtggctggac
     1081 caatgtaaat attgtcatga actatatccg taacctggat agtgaaacag gggcaatggt
     1141 gcgcctgctg gaagatggcg attagccatg tttaagggtt ccggttccac taggtacaat
     1201 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     1261 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     1321 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     1381 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     1441 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     1501 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     1561 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     1621 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     1681 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     1741 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     1801 cctccccgca tcgataccgt cgacctcgat cgagacctag aaaaacatgg agcaatcaca
     1861 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag
     1921 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct
     1981 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     2041 cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt ccctgattgg
     2101 cagaactaca caccagggcc agggatcaga tatccactga cctttggatg gtgctacaag
     2161 ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa cacccgcttg
     2221 ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtatt agagtggagg
     2281 tttgacagcc gcctagcatt tcatcacatg gcccgagagc tgcatccgga ctgtactggg
     2341 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     2401 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     2461 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagc
     2521 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2581 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2641 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2701 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2761 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2821 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2881 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2941 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3001 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3061 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3121 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3181 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3241 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3301 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3361 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3421 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3481 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3541 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3601 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3661 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3721 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3781 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3841 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3901 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3961 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4021 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4081 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4141 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4201 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4261 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4321 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     4381 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     4441 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     4501 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     4561 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     4621 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     4681 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     4741 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     4801 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     4861 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     4921 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     4981 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     5041 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     5101 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc
     5161 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     5221 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     5281 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     5341 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     5401 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     5461 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     5521 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     5581 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     5641 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     5701 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     5761 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     5821 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc
     5881 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     5941 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     6001 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     6061 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     6121 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     6181 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     6241 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     6301 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     6361 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     6421 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     6481 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     6541 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     6601 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
     6661 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
     6721 cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta
     6781 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     6841 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     6901 attacaggga cagcagagat ccagtttggt tagtaccggg cccgctctag ccctgcagga
     6961 actgtttaaa ctgtgatccg acgcgccatc tctaggcccg cgccggcccc ctcgcacgga
     7021 cttgtgggag aagctcggct actcccctgc cccggttaat ttgcatataa tatttcctag
     7081 taactataga ggcttaatgt gcgataaaag acagataatc tgttcttttt aatactagct
     7141 acattttaca tgataggctt ggatttctat aagagataca aatactaaat tattatttta
     7201 aaaaacagca caaaaggaaa ctcaccctaa ctgtaaagta attgtgtgtt ttgagactat
     7261 aagtatccct tggagaacca ccttgttgga tgggtctaaa ggcggcgggg tttgagagct
     7321 aagcagaaag ctgcatagca agttcaaata aggctagtcc gtacacaact tgaaaaagtg
     7381 gcagccgagt cggctgcttt ttttgctcga atctacactc agctatggcg cgccccaagg
     7441 tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     7501 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
     7561 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     7621 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
     7681 tgtggaaagc cagaaacatg gctcaagcgc gaacccgact ggtttcagag ctaagcacaa
     7741 gagtgcatag caagttgaaa taaggctagt ccgtttacaa cttgaaaaag tggcacccga
     7801 gtcgggtgct ttttttgttt ggcgcgccta tggcttaatt aacatggcgg ccgcattcta
     7861 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
     7921 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
     7981 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
     8041 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
     8101 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
     8161 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
     8221 gggtgggtcc gggggcgggc tcaggggcgg
//
