LOCUS       sequence-352702-        7660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Same as pLL3.3;U6(BstXI)-XhoI-chimeric RNA;PGK-Cre but XhoI stuffer 
            removed, expresses neomycin gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7660)
  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 7660)
  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-sgNeo1 (opti)
FEATURES             Location/Qualifiers
     source          1..7660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..34
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             86..106
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             107..1135
                     /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    1191..1779
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1244..1264)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1662..1673)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1782..1798)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1783..1799)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2308..2488
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2550..3138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2630..2649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3309..4100,4101..4169))
                     /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     3932..3951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4170..4274)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4349..4368)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4540..4919
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4921..5119
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5074..5094
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5137..5317
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5364..5489
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5986..6219
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6404..6448
                     /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    6746..6863
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6945..7193
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6945..6965
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7116..7135
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(7388..7411)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
       61 ttcctcatct ccgggacccg ggatgcccaa gaagaagagg aaggtgtcca atttactgac
      121 cgtacaccaa aatttgcctg cattaccggt cgatgcaacg agtgatgagg ttcgcaagaa
      181 cctgatggac atgttcaggg atcgccaggc gttttctgag catacctgga aaatgcttct
      241 gtccgtttgc cggtcgtggg cggcatggtg caagttgaat aaccggaaat ggtttcccgc
      301 agaacctgaa gatgttcgcg attatcttct atatcttcag gcgcgcggtc tggcagtaaa
      361 aactatccag caacatttgg gccagctaaa catgcttcat cgtcggtccg ggctgccacg
      421 accaagtgac agcaatgctg tttcactggt tatgcggcgg atccgaaaag aaaacgttga
      481 tgccggtgaa cgtgcaaaac aggctctagc gttcgaacgc actgatttcg accaggttcg
      541 ttcactcatg gaaaatagcg atcgctgcca ggatatacgt aatctggcat ttctggggat
      601 tgcttataac accctgttac gtatagccga aattgccagg atcagggtta aagatatctc
      661 acgtactgac ggtgggagaa tgttaatcca tattggcaga acgaaaacgc tggttagcac
      721 cgcaggtgta gagaaggcac ttagcctggg ggtaactaaa ctggtcgagc gatggatttc
      781 cgtctctggt gtagctgatg atccgaataa ctacctgttt tgccgggtca gaaaaaatgg
      841 tgttgccgcg ccatctgcca ccagccagct atcaactcgc gccctggaag ggatttttga
      901 agcaactcat cgattgattt acggcgctaa ggatgactct ggtcagagat acctggcctg
      961 gtctggacac agtgcccgtg tcggagccgc gcgagatatg gcccgcgctg gagtttcaat
     1021 accggagatc atgcaagctg gtggctggac caatgtaaat attgtcatga actatatccg
     1081 taacctggat agtgaaacag gggcaatggt gcgcctgctg gaagatggcg attagccatg
     1141 tttaagggtt ccggttccac taggtacaat tcgatatcaa gcttatcgat aatcaacctc
     1201 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     1261 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     1321 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     1381 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     1441 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     1501 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     1561 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     1621 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     1681 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     1741 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgat
     1801 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat
     1861 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta
     1921 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     1981 gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc
     2041 taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga
     2101 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa
     2161 gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat
     2221 gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg
     2281 gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg
     2341 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     2401 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     2461 ttttagtcag tgtggaaaat ctctagcagc atgtgagcaa aaggccagca aaaggccagg
     2521 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2581 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2641 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2701 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2761 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2821 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2881 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2941 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3001 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3061 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3121 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3181 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3241 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3301 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3361 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3421 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3481 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3541 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3601 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3661 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3721 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3781 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3841 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3901 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3961 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4021 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4081 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4141 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4201 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4261 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     4321 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4381 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     4441 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     4501 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     4561 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     4621 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     4681 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     4741 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     4801 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     4861 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     4921 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     4981 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     5041 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     5101 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
     5161 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5221 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     5281 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
     5341 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
     5401 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
     5461 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
     5521 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
     5581 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
     5641 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     5701 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     5761 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     5821 agcaagcggc cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
     5881 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     5941 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     6001 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     6061 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     6121 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     6181 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     6241 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     6301 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     6361 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     6421 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     6481 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     6541 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     6601 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag
     6661 agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc caattctgca
     6721 gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg
     6781 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     6841 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttggt
     6901 tagtaccggg cccgctctag ccctgcagga actgtttaaa ctgtgagggc ctatttccca
     6961 tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt
     7021 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
     7081 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     7141 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggaccaaac accgtcatgg
     7201 ctgatgcaat gcgggtttaa gagctatgct ggaaacagca tagcaagttt aaataaggct
     7261 agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgttt ggcgcgccta
     7321 tggcttaatt aacatggcgg ccgcattcta ccgggtaggg gaggcgcttt tcccaaggca
     7381 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
     7441 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
     7501 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
     7561 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
     7621 caccgctgag caatggaagc gggtaggcct ttggggcagc
//
