LOCUS       sequence-363976-        7770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Same as pLL3.3;U6(BstXI)-XhoI-chimeric RNA;PGK-Cre but XhoI stuffer 
            removed, expresses Cebpd gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7770)
  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 7770)
  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: hu6-sgCebpd_v1 (opti)
FEATURES             Location/Qualifiers
     source          1..7770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     76..95
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             147..167
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             168..1196
                     /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    1252..1840
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1305..1325)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1723..1734)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1843..1859)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1844..1860)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2369..2549
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2611..3199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2691..2710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3370..4161,4162..4230))
                     /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     3993..4012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4231..4335)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4410..4429)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4601..4980
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4982..5180
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5135..5155
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5198..5378
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5425..5550
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6047..6280
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6465..6509
                     /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    6807..6924
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7006..7254
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7006..7026
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7177..7196
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        join(7414..7770,1..143)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7449..7472)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt
       61 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
      121 cttcctcatc tccgggaccc gggatgccca agaagaagag gaaggtgtcc aatttactga
      181 ccgtacacca aaatttgcct gcattaccgg tcgatgcaac gagtgatgag gttcgcaaga
      241 acctgatgga catgttcagg gatcgccagg cgttttctga gcatacctgg aaaatgcttc
      301 tgtccgtttg ccggtcgtgg gcggcatggt gcaagttgaa taaccggaaa tggtttcccg
      361 cagaacctga agatgttcgc gattatcttc tatatcttca ggcgcgcggt ctggcagtaa
      421 aaactatcca gcaacatttg ggccagctaa acatgcttca tcgtcggtcc gggctgccac
      481 gaccaagtga cagcaatgct gtttcactgg ttatgcggcg gatccgaaaa gaaaacgttg
      541 atgccggtga acgtgcaaaa caggctctag cgttcgaacg cactgatttc gaccaggttc
      601 gttcactcat ggaaaatagc gatcgctgcc aggatatacg taatctggca tttctgggga
      661 ttgcttataa caccctgtta cgtatagccg aaattgccag gatcagggtt aaagatatct
      721 cacgtactga cggtgggaga atgttaatcc atattggcag aacgaaaacg ctggttagca
      781 ccgcaggtgt agagaaggca cttagcctgg gggtaactaa actggtcgag cgatggattt
      841 ccgtctctgg tgtagctgat gatccgaata actacctgtt ttgccgggtc agaaaaaatg
      901 gtgttgccgc gccatctgcc accagccagc tatcaactcg cgccctggaa gggatttttg
      961 aagcaactca tcgattgatt tacggcgcta aggatgactc tggtcagaga tacctggcct
     1021 ggtctggaca cagtgcccgt gtcggagccg cgcgagatat ggcccgcgct ggagtttcaa
     1081 taccggagat catgcaagct ggtggctgga ccaatgtaaa tattgtcatg aactatatcc
     1141 gtaacctgga tagtgaaaca ggggcaatgg tgcgcctgct ggaagatggc gattagccat
     1201 gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga taatcaacct
     1261 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     1321 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     1381 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     1441 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     1501 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     1561 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     1621 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     1681 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     1741 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     1801 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga
     1861 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     1921 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     1981 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     2041 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     2101 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
     2161 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
     2221 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
     2281 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
     2341 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
     2401 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     2461 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     2521 cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag
     2581 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2641 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2701 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2761 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2821 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2881 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2941 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3001 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3061 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3121 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3181 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3241 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3301 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3361 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3421 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3481 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3541 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3601 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3661 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3721 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3781 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3841 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3901 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3961 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4021 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4081 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4141 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4201 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4261 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4321 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     4381 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4441 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     4501 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     4561 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
     4621 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     4681 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     4741 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     4801 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     4861 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     4921 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     4981 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     5041 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     5101 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     5161 tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat
     5221 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     5281 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     5341 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     5401 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     5461 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     5521 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     5581 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     5641 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     5701 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     5761 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     5821 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     5881 cagcaagcgg ccggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     5941 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     6001 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     6061 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     6121 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     6181 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     6241 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     6301 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     6361 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     6421 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     6481 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     6541 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     6601 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     6661 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     6721 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     6781 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     6841 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     6901 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     6961 ttagtaccgg gcccgctcta gccctgcagg aactgtttaa actgtgaggg cctatttccc
     7021 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tggaattaat
     7081 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     7141 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
     7201 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgctcaa
     7261 gcgcgaaccc gactggttta agagctatgc tggaaacagc atagcaagtt taaataaggc
     7321 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt tggcgcgcct
     7381 atggcttaat taacatggcg gccgcattct accgggtagg ggaggcgctt ttcccaaggc
     7441 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
     7501 ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc
     7561 ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg
     7621 cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca
     7681 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct
     7741 ccttcgcttt ctgggctcag aggctgggaa
//
