LOCUS       sequence-352715-        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 Cebpb 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-sgCebpb_v1 (opti)
FEATURES             Location/Qualifiers
     source          1..7770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..593
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(58..78)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(476..487)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(596..612)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(597..613)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1122..1302
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(1364..1952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1444..1463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2123..2914,2915..2983))
                     /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     2746..2765
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2984..3088)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3163..3182)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3354..3733
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3735..3933
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3888..3908
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3951..4131
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4178..4303
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4800..5033
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5218..5262
                     /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    5560..5677
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5759..6007
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5759..5779
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5930..5949
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        6167..6666
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6202..6225)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6599..6618
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6670..6690
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6691..7719
                     /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 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
       61 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
      121 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
      181 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
      241 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
      301 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
      361 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
      421 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
      481 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
      541 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
      601 ccgtcgacct cgatcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag
      661 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag
      721 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
      781 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc
      841 ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag
      901 ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
      961 aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc
     1021 tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag
     1081 catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc
     1141 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     1201 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     1261 gatccctcag acccttttag tcagtgtgga aaatctctag cagcatgtga gcaaaaggcc
     1321 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1381 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1441 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1501 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1561 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1621 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1681 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1741 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1801 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1861 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1921 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1981 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2041 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2101 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2161 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2221 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     2281 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2341 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2401 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2461 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2521 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2581 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2641 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2701 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     2761 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     2821 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     2881 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2941 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3001 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3061 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     3121 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     3181 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     3241 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     3301 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
     3361 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     3421 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     3481 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     3541 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     3601 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     3661 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     3721 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     3781 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     3841 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     3901 taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact gggtctctct
     3961 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     4021 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     4081 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
     4141 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
     4201 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     4261 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     4321 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     4381 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     4441 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     4501 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     4561 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     4621 taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg aggagatatg
     4681 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     4741 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     4801 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     4861 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     4921 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     4981 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     5041 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     5101 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     5161 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     5221 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     5281 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     5341 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     5401 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     5461 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     5521 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     5581 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     5641 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     5701 agatccagtt tggttagtac cgggcccgct ctagccctgc aggaactgtt taaactgtga
     5761 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     5821 aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     5881 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
     5941 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
     6001 aaacaccgat gggtctaaag gcggcggggt ttaagagcta tgctggaaac agcatagcaa
     6061 gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
     6121 gtttggcgcg cctatggctt aattaacatg gcggccgcat tctaccgggt aggggaggcg
     6181 cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac
     6241 acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg
     6301 ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc
     6361 gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc
     6421 gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa
     6481 tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc
     6541 gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca
     6601 ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccggga
     6661 cccgggatgc ccaagaagaa gaggaaggtg tccaatttac tgaccgtaca ccaaaatttg
     6721 cctgcattac cggtcgatgc aacgagtgat gaggttcgca agaacctgat ggacatgttc
     6781 agggatcgcc aggcgttttc tgagcatacc tggaaaatgc ttctgtccgt ttgccggtcg
     6841 tgggcggcat ggtgcaagtt gaataaccgg aaatggtttc ccgcagaacc tgaagatgtt
     6901 cgcgattatc ttctatatct tcaggcgcgc ggtctggcag taaaaactat ccagcaacat
     6961 ttgggccagc taaacatgct tcatcgtcgg tccgggctgc cacgaccaag tgacagcaat
     7021 gctgtttcac tggttatgcg gcggatccga aaagaaaacg ttgatgccgg tgaacgtgca
     7081 aaacaggctc tagcgttcga acgcactgat ttcgaccagg ttcgttcact catggaaaat
     7141 agcgatcgct gccaggatat acgtaatctg gcatttctgg ggattgctta taacaccctg
     7201 ttacgtatag ccgaaattgc caggatcagg gttaaagata tctcacgtac tgacggtggg
     7261 agaatgttaa tccatattgg cagaacgaaa acgctggtta gcaccgcagg tgtagagaag
     7321 gcacttagcc tgggggtaac taaactggtc gagcgatgga tttccgtctc tggtgtagct
     7381 gatgatccga ataactacct gttttgccgg gtcagaaaaa atggtgttgc cgcgccatct
     7441 gccaccagcc agctatcaac tcgcgccctg gaagggattt ttgaagcaac tcatcgattg
     7501 atttacggcg ctaaggatga ctctggtcag agatacctgg cctggtctgg acacagtgcc
     7561 cgtgtcggag ccgcgcgaga tatggcccgc gctggagttt caataccgga gatcatgcaa
     7621 gctggtggct ggaccaatgt aaatattgtc atgaactata tccgtaacct ggatagtgaa
     7681 acaggggcaa tggtgcgcct gctggaagat ggcgattagc catgtttaag ggttccggtt
     7741 ccactaggta caattcgata tcaagcttat
//
