LOCUS       sequence-352705-        7769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Same as pLL3.3;U6(BstXI)-XhoI-chimeric RNA;PGK-Cre but XhoI stuffer 
            removed, expresses Lkb1 gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7769)
  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 7769)
  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-sgLkb1_v2 (opti)
FEATURES             Location/Qualifiers
     source          1..7769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..66
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             118..138
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             139..1167
                     /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    1223..1811
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1276..1296)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1694..1705)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1814..1830)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1815..1831)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2340..2520
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2582..3170)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2662..2681)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3341..4132,4133..4201))
                     /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     3964..3983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4202..4306)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4381..4400)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4572..4951
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4953..5151
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5106..5126
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5169..5349
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5396..5521
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6018..6251
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6436..6480
                     /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    6778..6895
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6977..7225
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6977..6997
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7148..7167
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        join(7385..7769,1..114)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7420..7443)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt
       61 caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggacc cgggatgccc
      121 aagaagaaga ggaaggtgtc caatttactg accgtacacc aaaatttgcc tgcattaccg
      181 gtcgatgcaa cgagtgatga ggttcgcaag aacctgatgg acatgttcag ggatcgccag
      241 gcgttttctg agcatacctg gaaaatgctt ctgtccgttt gccggtcgtg ggcggcatgg
      301 tgcaagttga ataaccggaa atggtttccc gcagaacctg aagatgttcg cgattatctt
      361 ctatatcttc aggcgcgcgg tctggcagta aaaactatcc agcaacattt gggccagcta
      421 aacatgcttc atcgtcggtc cgggctgcca cgaccaagtg acagcaatgc tgtttcactg
      481 gttatgcggc ggatccgaaa agaaaacgtt gatgccggtg aacgtgcaaa acaggctcta
      541 gcgttcgaac gcactgattt cgaccaggtt cgttcactca tggaaaatag cgatcgctgc
      601 caggatatac gtaatctggc atttctgggg attgcttata acaccctgtt acgtatagcc
      661 gaaattgcca ggatcagggt taaagatatc tcacgtactg acggtgggag aatgttaatc
      721 catattggca gaacgaaaac gctggttagc accgcaggtg tagagaaggc acttagcctg
      781 ggggtaacta aactggtcga gcgatggatt tccgtctctg gtgtagctga tgatccgaat
      841 aactacctgt tttgccgggt cagaaaaaat ggtgttgccg cgccatctgc caccagccag
      901 ctatcaactc gcgccctgga agggattttt gaagcaactc atcgattgat ttacggcgct
      961 aaggatgact ctggtcagag atacctggcc tggtctggac acagtgcccg tgtcggagcc
     1021 gcgcgagata tggcccgcgc tggagtttca ataccggaga tcatgcaagc tggtggctgg
     1081 accaatgtaa atattgtcat gaactatatc cgtaacctgg atagtgaaac aggggcaatg
     1141 gtgcgcctgc tggaagatgg cgattagcca tgtttaaggg ttccggttcc actaggtaca
     1201 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     1261 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     1321 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     1381 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     1441 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     1501 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     1561 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     1621 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     1681 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     1741 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     1801 cgcctccccg catcgatacc gtcgacctcg atcgagacct agaaaaacat ggagcaatca
     1861 caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg
     1921 aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag
     1981 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     2041 aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt
     2101 ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca
     2161 agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct
     2221 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga
     2281 ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg
     2341 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     2401 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     2461 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     2521 gcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2581 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2641 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2701 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2761 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2821 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2881 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2941 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3001 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3061 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3121 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3181 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3241 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3301 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3361 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3421 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3481 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3541 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3601 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3661 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3721 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3781 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3841 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3901 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3961 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4021 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4081 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4141 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4201 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4261 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4321 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     4381 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     4441 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     4501 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     4561 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     4621 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     4681 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     4741 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     4801 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     4861 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     4921 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     4981 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     5041 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     5101 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg
     5161 cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     5221 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     5281 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     5341 tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc
     5401 tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt
     5461 gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt
     5521 cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg
     5581 aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc
     5641 agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca
     5701 accatccctt cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct
     5761 ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag acaagataga
     5821 ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccggccgct gatcttcaga
     5881 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     5941 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     6001 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     6061 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     6121 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     6181 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     6241 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     6301 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     6361 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     6421 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     6481 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     6541 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     6601 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
     6661 cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg
     6721 aacggatcgg cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt
     6781 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     6841 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     6901 ttattacagg gacagcagag atccagtttg gttagtaccg ggcccgctct agccctgcag
     6961 gaactgttta aactgtgagg gcctatttcc catgattcct tcatatttgc atatacgata
     7021 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
     7081 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     7141 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     7201 tatcttgtgg aaaggaccaa acaccgccac tctctgacct actccggttt aagagctatg
     7261 ctggaaacag catagcaagt ttaaataagg ctagtccgtt atcaacttga aaaagtggca
     7321 ccgagtcggt gcttttttgt ttggcgcgcc tatggcttaa ttaacatggc ggccgcattc
     7381 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     7441 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     7501 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     7561 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     7621 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     7681 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     7741 aggggtgggt ccgggggcgg gctcagggg
//
