LOCUS       sequence-352720-        8248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses  Cebpb_v2 (mU6), Cebpd_v2(hU6) gRNAs and Cre-recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8248)
  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 8248)
  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_v2-hu6-sgCebpd_v2
FEATURES             Location/Qualifiers
     source          1..8248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             79..259
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(321..909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(401..420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1080..1871,1872..1940))
                     /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     1703..1722
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1941..2045)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2120..2139)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2311..2690
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2692..2890
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2845..2865
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2908..3088
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3135..3260
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3757..3990
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4175..4219
                     /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    4517..4634
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     4947..4967
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     5192..5212
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5363..5382
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        5602..6101
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5637..5660)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6034..6053
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6105..6125
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6126..7154
                     /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    7210..7798
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7263..7283)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7681..7692)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7801..7817)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7802..7818)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag
       61 ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
      121 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
      181 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
      241 gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      301 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      361 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      421 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      481 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      541 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      601 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      661 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      721 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      781 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      841 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      901 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      961 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1021 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1081 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1141 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1201 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1261 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1321 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1381 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1441 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1501 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1561 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     1621 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     1681 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     1741 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     1801 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     1861 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     1921 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     1981 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2041 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     2101 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     2161 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     2221 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     2281 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     2341 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     2401 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     2461 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     2521 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     2581 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     2641 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     2701 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     2761 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     2821 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     2881 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
     2941 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     3001 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     3061 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     3121 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     3181 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     3241 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     3301 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     3361 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     3421 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     3481 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     3541 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     3601 ccggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     3661 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     3721 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     3781 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     3841 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     3901 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     3961 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     4021 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     4081 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     4141 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     4201 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     4261 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     4321 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     4381 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     4441 cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc agacaaatgg
     4501 cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
     4561 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
     4621 aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg ttagtaccgg
     4681 gcccgctcta gccctgcagg aactgtttaa actgtgatcc gacgcgccat ctctaggccc
     4741 gcgccggccc cctcgcacgg acttgtggga gaagctcggc tactcccctg ccccggttaa
     4801 tttgcatata atatttccta gtaactatag aggcttaatg tgcgataaaa gacagataat
     4861 ctgttctttt taatactagc tacattttac atgataggct tggatttcta taagagatac
     4921 aaatactaaa ttattatttt aaaaaacagc acaaaaggaa actcacccta actgtaaagt
     4981 aattgtgtgt tttgagacta taagtatccc ttggagaacc accttgttgg tgctcgccaa
     5041 tggccggctg tttgagagct aagcagaaag ctgcatagca agttcaaata aggctagtcc
     5101 gtacacaact tgaaaaagtg gcagccgagt cggctgcttt ttttgctcga atctacactc
     5161 agctatggcg cgccccaagg tcgggcagga agagggccta tttcccatga ttccttcata
     5221 tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac
     5281 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
     5341 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
     5401 tttcttggct ttatatatct tgtggaaagc cagaaacatg gtcgtacatg gcaggagtcg
     5461 gtttcagagc taagcacaag agtgcatagc aagttgaaat aaggctagtc cgtttacaac
     5521 ttgaaaaagt ggcacccgag tcgggtgctt tttttgtttg gcgcgcctat ggcttaatta
     5581 acatggcggc cgcattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
     5641 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
     5701 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
     5761 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
     5821 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
     5881 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
     5941 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
     6001 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
     6061 ctgccgcgct gttctcctct tcctcatctc cgggacccgg gatgcccaag aagaagagga
     6121 aggtgtccaa tttactgacc gtacaccaaa atttgcctgc attaccggtc gatgcaacga
     6181 gtgatgaggt tcgcaagaac ctgatggaca tgttcaggga tcgccaggcg ttttctgagc
     6241 atacctggaa aatgcttctg tccgtttgcc ggtcgtgggc ggcatggtgc aagttgaata
     6301 accggaaatg gtttcccgca gaacctgaag atgttcgcga ttatcttcta tatcttcagg
     6361 cgcgcggtct ggcagtaaaa actatccagc aacatttggg ccagctaaac atgcttcatc
     6421 gtcggtccgg gctgccacga ccaagtgaca gcaatgctgt ttcactggtt atgcggcgga
     6481 tccgaaaaga aaacgttgat gccggtgaac gtgcaaaaca ggctctagcg ttcgaacgca
     6541 ctgatttcga ccaggttcgt tcactcatgg aaaatagcga tcgctgccag gatatacgta
     6601 atctggcatt tctggggatt gcttataaca ccctgttacg tatagccgaa attgccagga
     6661 tcagggttaa agatatctca cgtactgacg gtgggagaat gttaatccat attggcagaa
     6721 cgaaaacgct ggttagcacc gcaggtgtag agaaggcact tagcctgggg gtaactaaac
     6781 tggtcgagcg atggatttcc gtctctggtg tagctgatga tccgaataac tacctgtttt
     6841 gccgggtcag aaaaaatggt gttgccgcgc catctgccac cagccagcta tcaactcgcg
     6901 ccctggaagg gatttttgaa gcaactcatc gattgattta cggcgctaag gatgactctg
     6961 gtcagagata cctggcctgg tctggacaca gtgcccgtgt cggagccgcg cgagatatgg
     7021 cccgcgctgg agtttcaata ccggagatca tgcaagctgg tggctggacc aatgtaaata
     7081 ttgtcatgaa ctatatccgt aacctggata gtgaaacagg ggcaatggtg cgcctgctgg
     7141 aagatggcga ttagccatgt ttaagggttc cggttccact aggtacaatt cgatatcaag
     7201 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     7261 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     7321 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     7381 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     7441 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     7501 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     7561 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     7621 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     7681 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     7741 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     7801 cgataccgtc gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     7861 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt
     7921 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     7981 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     8041 tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc agaactacac
     8101 accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     8161 tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt
     8221 gagcctgcat gggatggatg acccggag
//
