LOCUS       sequence-352706-        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_v1 (opti)
FEATURES             Location/Qualifiers
     source          1..7770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1035..1623
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1088..1108)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1506..1517)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1626..1642)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1627..1643)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2152..2332
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2394..2982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2474..2493)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3153..3944,3945..4013))
                     /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     3776..3795
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4014..4118)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4193..4212)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4384..4763
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4765..4963
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4918..4938
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4981..5161
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5208..5333
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5830..6063
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6248..6292
                     /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    6590..6707
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6789..7037
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6789..6809
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6960..6979
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        7197..7696
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7232..7255)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7629..7648
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7700..7720
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(7721..7770,1..979)
                     /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"
ORIGIN
        1 aacgagtgat gaggttcgca agaacctgat ggacatgttc agggatcgcc aggcgttttc
       61 tgagcatacc tggaaaatgc ttctgtccgt ttgccggtcg tgggcggcat ggtgcaagtt
      121 gaataaccgg aaatggtttc ccgcagaacc tgaagatgtt cgcgattatc ttctatatct
      181 tcaggcgcgc ggtctggcag taaaaactat ccagcaacat ttgggccagc taaacatgct
      241 tcatcgtcgg tccgggctgc cacgaccaag tgacagcaat gctgtttcac tggttatgcg
      301 gcggatccga aaagaaaacg ttgatgccgg tgaacgtgca aaacaggctc tagcgttcga
      361 acgcactgat ttcgaccagg ttcgttcact catggaaaat agcgatcgct gccaggatat
      421 acgtaatctg gcatttctgg ggattgctta taacaccctg ttacgtatag ccgaaattgc
      481 caggatcagg gttaaagata tctcacgtac tgacggtggg agaatgttaa tccatattgg
      541 cagaacgaaa acgctggtta gcaccgcagg tgtagagaag gcacttagcc tgggggtaac
      601 taaactggtc gagcgatgga tttccgtctc tggtgtagct gatgatccga ataactacct
      661 gttttgccgg gtcagaaaaa atggtgttgc cgcgccatct gccaccagcc agctatcaac
      721 tcgcgccctg gaagggattt ttgaagcaac tcatcgattg atttacggcg ctaaggatga
      781 ctctggtcag agatacctgg cctggtctgg acacagtgcc cgtgtcggag ccgcgcgaga
      841 tatggcccgc gctggagttt caataccgga gatcatgcaa gctggtggct ggaccaatgt
      901 aaatattgtc atgaactata tccgtaacct ggatagtgaa acaggggcaa tggtgcgcct
      961 gctggaagat ggcgattagc catgtttaag ggttccggtt ccactaggta caattcgata
     1021 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1081 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1141 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     1201 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     1261 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     1321 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     1381 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     1441 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     1501 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     1561 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     1621 cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat cacaagtagc
     1681 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg
     1741 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat
     1801 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     1861 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac
     1921 tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta
     1981 ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac
     2041 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac
     2101 agccgcctag catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc
     2161 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     2221 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     2281 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagcatgtga
     2341 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2401 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2461 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2521 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2581 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2641 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2701 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2761 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2821 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2881 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2941 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3001 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3061 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3121 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3181 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3241 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3301 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3361 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3421 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3481 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3541 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3601 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3661 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3721 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3781 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3841 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3901 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3961 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4021 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4081 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4141 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     4201 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     4261 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     4321 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     4381 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     4441 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     4501 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     4561 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     4621 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     4681 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     4741 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     4801 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     4861 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     4921 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact
     4981 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5041 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5101 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5161 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     5221 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     5281 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     5341 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     5401 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     5461 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     5521 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     5581 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     5641 aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg
     5701 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     5761 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     5821 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     5881 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     5941 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     6001 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     6061 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     6121 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     6181 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     6241 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     6301 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     6361 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     6421 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     6481 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     6541 ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa
     6601 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     6661 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     6721 gggacagcag agatccagtt tggttagtac cgggcccgct ctagccctgc aggaactgtt
     6781 taaactgtga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
     6841 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
     6901 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
     6961 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
     7021 ggaaaggacg aaacaccgtg ctcgcagatg ccgcccaggt ttaagagcta tgctggaaac
     7081 agcatagcaa gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     7141 gtgctttttt gtttggcgcg cctatggctt aattaacatg gcggccgcat tctaccgggt
     7201 aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac
     7261 ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca
     7321 accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa
     7381 gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct
     7441 cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg
     7501 cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg
     7561 gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg
     7621 aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc
     7681 atctccggga cccgggatgc ccaagaagaa gaggaaggtg tccaatttac tgaccgtaca
     7741 ccaaaatttg cctgcattac cggtcgatgc
//
