LOCUS       sequence-351960-        8249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses  Cebpa_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 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-sgCebpd_v2
FEATURES             Location/Qualifiers
     source          1..8249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             150..330
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(392..980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(472..491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1151..1942,1943..2011))
                     /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     1774..1793
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2012..2116)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2191..2210)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2382..2761
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2763..2961
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2916..2936
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2979..3159
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3206..3331
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3828..4061
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4246..4290
                     /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    4588..4705
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     5018..5038
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     5264..5284
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5435..5454
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        5674..6173
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5709..5732)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6106..6125
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6177..6197
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6198..7226
                     /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    7282..7870
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7335..7355)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7753..7764)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7873..7889)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7874..7890)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 aagaagccaa tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg
       61 atgacccgga gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca
      121 tggcccgaga gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct
      181 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      241 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
      301 ccttttagtc agtgtggaaa atctctagca gcatgtgagc aaaaggccag caaaaggcca
      361 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      421 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      481 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      541 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      601 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      661 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      721 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      781 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      841 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
      901 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
      961 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1021 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1081 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1141 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1201 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     1261 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     1321 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     1381 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     1441 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     1501 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     1561 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     1621 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     1681 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     1741 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     1801 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     1861 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     1921 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     1981 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     2041 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     2101 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     2161 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2221 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     2281 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     2341 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
     2401 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     2461 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     2521 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     2581 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     2641 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     2701 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     2761 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     2821 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     2881 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     2941 gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg ttagaccaga
     3001 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     3061 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     3121 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     3181 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     3241 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     3301 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     3361 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     3421 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     3481 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     3541 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     3601 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     3661 acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     3721 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     3781 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     3841 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     3901 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     3961 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     4021 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     4081 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     4141 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     4201 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     4261 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     4321 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     4381 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     4441 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     4501 agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc gccaattctg
     4561 cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
     4621 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
     4681 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg
     4741 gttagtaccg ggcccgctct agccctgcag gaactgttta aactgtgatc cgacgcgcca
     4801 tctctaggcc cgcgccggcc ccctcgcacg gacttgtggg agaagctcgg ctactcccct
     4861 gccccggtta atttgcatat aatatttcct agtaactata gaggcttaat gtgcgataaa
     4921 agacagataa tctgttcttt ttaatactag ctacatttta catgataggc ttggatttct
     4981 ataagagata caaatactaa attattattt taaaaaacag cacaaaagga aactcaccct
     5041 aactgtaaag taattgtgtg ttttgagact ataagtatcc cttggagaac caccttgttg
     5101 gcatcagcgc ctacatcgac cgtttgagag ctaagcagaa agctgcatag caagttcaaa
     5161 taaggctagt ccgtacacaa cttgaaaaag tggcagccga gtcggctgct ttttttgctc
     5221 gaatctacac tcagctatgg cgcgccccaa ggtcgggcag gaagagggcc tatttcccat
     5281 gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt
     5341 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
     5401 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
     5461 aaagtatttc gatttcttgg ctttatatat cttgtggaaa gccagaaaca tggtcgtaca
     5521 tggcaggagt cggtttcaga gctaagcaca agagtgcata gcaagttgaa ataaggctag
     5581 tccgtttaca acttgaaaaa gtggcacccg agtcgggtgc tttttttgtt tggcgcgcct
     5641 atggcttaat taacatggcg gccgcattct accgggtagg ggaggcgctt ttcccaaggc
     5701 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
     5761 ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc
     5821 ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg
     5881 cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca
     5941 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct
     6001 ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg
     6061 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
     6121 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggaccc gggatgccca
     6181 agaagaagag gaaggtgtcc aatttactga ccgtacacca aaatttgcct gcattaccgg
     6241 tcgatgcaac gagtgatgag gttcgcaaga acctgatgga catgttcagg gatcgccagg
     6301 cgttttctga gcatacctgg aaaatgcttc tgtccgtttg ccggtcgtgg gcggcatggt
     6361 gcaagttgaa taaccggaaa tggtttcccg cagaacctga agatgttcgc gattatcttc
     6421 tatatcttca ggcgcgcggt ctggcagtaa aaactatcca gcaacatttg ggccagctaa
     6481 acatgcttca tcgtcggtcc gggctgccac gaccaagtga cagcaatgct gtttcactgg
     6541 ttatgcggcg gatccgaaaa gaaaacgttg atgccggtga acgtgcaaaa caggctctag
     6601 cgttcgaacg cactgatttc gaccaggttc gttcactcat ggaaaatagc gatcgctgcc
     6661 aggatatacg taatctggca tttctgggga ttgcttataa caccctgtta cgtatagccg
     6721 aaattgccag gatcagggtt aaagatatct cacgtactga cggtgggaga atgttaatcc
     6781 atattggcag aacgaaaacg ctggttagca ccgcaggtgt agagaaggca cttagcctgg
     6841 gggtaactaa actggtcgag cgatggattt ccgtctctgg tgtagctgat gatccgaata
     6901 actacctgtt ttgccgggtc agaaaaaatg gtgttgccgc gccatctgcc accagccagc
     6961 tatcaactcg cgccctggaa gggatttttg aagcaactca tcgattgatt tacggcgcta
     7021 aggatgactc tggtcagaga tacctggcct ggtctggaca cagtgcccgt gtcggagccg
     7081 cgcgagatat ggcccgcgct ggagtttcaa taccggagat catgcaagct ggtggctgga
     7141 ccaatgtaaa tattgtcatg aactatatcc gtaacctgga tagtgaaaca ggggcaatgg
     7201 tgcgcctgct ggaagatggc gattagccat gtttaagggt tccggttcca ctaggtacaa
     7261 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     7321 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     7381 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     7441 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     7501 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     7561 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     7621 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     7681 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     7741 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     7801 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     7861 gcctccccgc atcgataccg tcgacctcga tcgagaccta gaaaaacatg gagcaatcac
     7921 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
     7981 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     8041 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     8101 acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg
     8161 gcagaactac acaccagggc cagggatcag atatccactg acctttggat ggtgctacaa
     8221 gctagtacca gttgagcaag agaaggtag
//
