LOCUS       sequence-352708-        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 Cebpa 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-sgCebpa_v2 (opti)
FEATURES             Location/Qualifiers
     source          1..7770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             41..61
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             62..1090
                     /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    1146..1734
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1199..1219)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1617..1628)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1737..1753)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1738..1754)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2263..2443
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2505..3093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2585..2604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3264..4055,4056..4124))
                     /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     3887..3906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4125..4229)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4304..4323)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4495..4874
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4876..5074
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5029..5049
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5092..5272
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5319..5444
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5941..6174
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6359..6403
                     /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    6701..6818
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6900..7148
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6900..6920
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7071..7090
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        join(7308..7770,1..37)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7343..7366)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7740..7759
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
ORIGIN
        1 cgcgctgttc tcctcttcct catctccggg acccgggatg cccaagaaga agaggaaggt
       61 gtccaattta ctgaccgtac accaaaattt gcctgcatta ccggtcgatg caacgagtga
      121 tgaggttcgc aagaacctga tggacatgtt cagggatcgc caggcgtttt ctgagcatac
      181 ctggaaaatg cttctgtccg tttgccggtc gtgggcggca tggtgcaagt tgaataaccg
      241 gaaatggttt cccgcagaac ctgaagatgt tcgcgattat cttctatatc ttcaggcgcg
      301 cggtctggca gtaaaaacta tccagcaaca tttgggccag ctaaacatgc ttcatcgtcg
      361 gtccgggctg ccacgaccaa gtgacagcaa tgctgtttca ctggttatgc ggcggatccg
      421 aaaagaaaac gttgatgccg gtgaacgtgc aaaacaggct ctagcgttcg aacgcactga
      481 tttcgaccag gttcgttcac tcatggaaaa tagcgatcgc tgccaggata tacgtaatct
      541 ggcatttctg gggattgctt ataacaccct gttacgtata gccgaaattg ccaggatcag
      601 ggttaaagat atctcacgta ctgacggtgg gagaatgtta atccatattg gcagaacgaa
      661 aacgctggtt agcaccgcag gtgtagagaa ggcacttagc ctgggggtaa ctaaactggt
      721 cgagcgatgg atttccgtct ctggtgtagc tgatgatccg aataactacc tgttttgccg
      781 ggtcagaaaa aatggtgttg ccgcgccatc tgccaccagc cagctatcaa ctcgcgccct
      841 ggaagggatt tttgaagcaa ctcatcgatt gatttacggc gctaaggatg actctggtca
      901 gagatacctg gcctggtctg gacacagtgc ccgtgtcgga gccgcgcgag atatggcccg
      961 cgctggagtt tcaataccgg agatcatgca agctggtggc tggaccaatg taaatattgt
     1021 catgaactat atccgtaacc tggatagtga aacaggggca atggtgcgcc tgctggaaga
     1081 tggcgattag ccatgtttaa gggttccggt tccactaggt acaattcgat atcaagctta
     1141 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     1201 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     1261 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     1321 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     1381 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     1441 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     1501 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     1561 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     1621 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     1681 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     1741 accgtcgacc tcgatcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     1801 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     1861 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     1921 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc
     1981 cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca
     2041 gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     2101 caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc
     2161 ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta
     2221 gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac
     2281 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     2341 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     2401 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagcatgtg agcaaaaggc
     2461 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2521 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2581 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2641 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2701 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2761 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2821 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2881 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2941 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3001 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3061 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3121 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3181 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3241 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3301 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3361 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3421 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3481 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3541 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3601 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3661 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3721 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3781 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3841 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3901 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3961 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4021 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4081 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4141 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4201 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4261 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     4321 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     4381 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     4441 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
     4501 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     4561 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     4621 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     4681 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4741 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4801 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4861 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     4921 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     4981 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     5041 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
     5101 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     5161 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     5221 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     5281 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     5341 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     5401 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     5461 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     5521 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     5581 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     5641 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     5701 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     5761 gtaagaccac cgcacagcaa gcggccggcc gctgatcttc agacctggag gaggagatat
     5821 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     5881 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     5941 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     6001 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     6061 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     6121 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     6181 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     6241 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     6301 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     6361 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     6421 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     6481 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     6541 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     6601 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg
     6661 tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     6721 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     6781 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     6841 gagatccagt ttggttagta ccgggcccgc tctagccctg caggaactgt ttaaactgtg
     6901 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
     6961 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
     7021 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
     7081 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     7141 gaaacaccgc atcagcgcct acatcgaccg tttaagagct atgctggaaa cagcatagca
     7201 agtttaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     7261 tgtttggcgc gcctatggct taattaacat ggcggccgca ttctaccggg taggggaggc
     7321 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
     7381 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
     7441 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
     7501 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
     7561 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
     7621 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
     7681 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
     7741 attctgcacg cttcaaaagc gcacgtctgc
//
