LOCUS       sequence-352704-        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_IW (opti)
FEATURES             Location/Qualifiers
     source          1..7769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(395..406)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(515..531)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(516..532)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1041..1221
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(1283..1871)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1363..1382)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2042..2833,2834..2902))
                     /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     2665..2684
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2903..3007)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3082..3101)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3273..3652
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3654..3852
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3807..3827
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3870..4050
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4097..4222
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4719..4952
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5137..5181
                     /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    5479..5596
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5678..5926
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5678..5698
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5849..5868
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        6085..6584
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6120..6143)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6517..6536
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6588..6608
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6609..7637
                     /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    join(7693..7769,1..512)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7746..7766)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
       61 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
      121 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
      181 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
      241 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
      301 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
      361 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
      421 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
      481 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc
      541 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct
      601 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa
      661 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
      721 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca
      781 cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc agatatccac
      841 tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca
      901 atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      961 agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag
     1021 agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     1081 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     1141 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     1201 cagtgtggaa aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1261 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1321 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1381 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1441 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1501 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1561 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1621 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1681 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     1741 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1801 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1861 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1921 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     1981 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2041 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2101 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2161 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2221 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2281 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2341 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2401 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     2461 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     2521 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     2581 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2641 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     2701 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     2761 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     2821 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     2881 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     2941 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3001 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     3061 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     3121 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     3181 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     3241 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     3301 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     3361 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     3421 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     3481 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     3541 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     3601 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     3661 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     3721 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     3781 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     3841 ctatataagc agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct
     3901 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     3961 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     4021 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     4081 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     4141 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     4201 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     4261 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     4321 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     4381 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     4441 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     4501 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     4561 ggccggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     4621 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     4681 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     4741 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     4801 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     4861 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     4921 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     4981 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     5041 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     5101 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     5161 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     5221 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     5281 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     5341 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     5401 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
     5461 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     5521 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     5581 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttagtacc
     5641 gggcccgctc tagccctgca ggaactgttt aaactgtgag ggcctatttc ccatgattcc
     5701 ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt
     5761 aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt
     5821 tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta
     5881 tttcgatttc ttggctttat atatcttgtg gaaaggacca aacaccgtgg tgggccgcag
     5941 tcacaagttt aagagctatg ctggaaacag catagcaagt ttaaataagg ctagtccgtt
     6001 atcaacttga aaaagtggca ccgagtcggt gcttttttgt ttggcgcgcc tatggcttaa
     6061 ttaacatggc ggccgcattc taccgggtag gggaggcgct tttcccaagg cagtctggag
     6121 catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca
     6181 cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc
     6241 caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca
     6301 ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg
     6361 agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt
     6421 tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg
     6481 gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca
     6541 cgtctgccgc gctgttctcc tcttcctcat ctccgggacc cgggatgccc aagaagaaga
     6601 ggaaggtgtc caatttactg accgtacacc aaaatttgcc tgcattaccg gtcgatgcaa
     6661 cgagtgatga ggttcgcaag aacctgatgg acatgttcag ggatcgccag gcgttttctg
     6721 agcatacctg gaaaatgctt ctgtccgttt gccggtcgtg ggcggcatgg tgcaagttga
     6781 ataaccggaa atggtttccc gcagaacctg aagatgttcg cgattatctt ctatatcttc
     6841 aggcgcgcgg tctggcagta aaaactatcc agcaacattt gggccagcta aacatgcttc
     6901 atcgtcggtc cgggctgcca cgaccaagtg acagcaatgc tgtttcactg gttatgcggc
     6961 ggatccgaaa agaaaacgtt gatgccggtg aacgtgcaaa acaggctcta gcgttcgaac
     7021 gcactgattt cgaccaggtt cgttcactca tggaaaatag cgatcgctgc caggatatac
     7081 gtaatctggc atttctgggg attgcttata acaccctgtt acgtatagcc gaaattgcca
     7141 ggatcagggt taaagatatc tcacgtactg acggtgggag aatgttaatc catattggca
     7201 gaacgaaaac gctggttagc accgcaggtg tagagaaggc acttagcctg ggggtaacta
     7261 aactggtcga gcgatggatt tccgtctctg gtgtagctga tgatccgaat aactacctgt
     7321 tttgccgggt cagaaaaaat ggtgttgccg cgccatctgc caccagccag ctatcaactc
     7381 gcgccctgga agggattttt gaagcaactc atcgattgat ttacggcgct aaggatgact
     7441 ctggtcagag atacctggcc tggtctggac acagtgcccg tgtcggagcc gcgcgagata
     7501 tggcccgcgc tggagtttca ataccggaga tcatgcaagc tggtggctgg accaatgtaa
     7561 atattgtcat gaactatatc cgtaacctgg atagtgaaac aggggcaatg gtgcgcctgc
     7621 tggaagatgg cgattagcca tgtttaaggg ttccggttcc actaggtaca attcgatatc
     7681 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     7741 aactatgttg ctccttttac gctatgtgg
//
