LOCUS       sequence-352716-        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 Cebpb 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-sgCebpb_v2 (opti)
FEATURES             Location/Qualifiers
     source          1..7769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4..24)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(422..433)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(542..558)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(543..559)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1068..1248
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(1310..1898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1390..1409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2069..2860,2861..2929))
                     /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     2692..2711
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2930..3034)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3109..3128)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3300..3679
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3681..3879
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3834..3854
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3897..4077
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4124..4249
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4746..4979
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5164..5208
                     /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    5506..5623
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5705..5953
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5705..5725
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5876..5895
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        6112..6611
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6147..6170)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6544..6563
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6615..6635
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6636..7664
                     /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(7720..7769,1..539)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
ORIGIN
        1 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
       61 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
      121 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
      181 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
      241 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
      301 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
      361 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
      421 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
      481 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
      541 tcgataccgt cgacctcgat cgagacctag aaaaacatgg agcaatcaca agtagcaata
      601 cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt
      661 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
      721 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
      781 atatccttga tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca
      841 caccagggcc agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag
      901 ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg
      961 tgagcctgca tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc
     1021 gcctagcatt tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt
     1081 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     1141 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     1201 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagc atgtgagcaa
     1261 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1321 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1381 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1441 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1501 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1561 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1621 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1681 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1741 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1801 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1861 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1921 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1981 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2041 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2101 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2161 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2221 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2281 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2341 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2401 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2461 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2521 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2581 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2641 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2701 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2761 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2821 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2881 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2941 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3001 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     3061 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     3121 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     3181 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     3241 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
     3301 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     3361 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     3421 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     3481 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     3541 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     3601 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     3661 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     3721 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     3781 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     3841 gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt
     3901 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     3961 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     4021 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     4081 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     4141 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     4201 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     4261 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     4321 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     4381 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     4441 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     4501 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     4561 caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga
     4621 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     4681 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     4741 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     4801 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     4861 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     4921 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     4981 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     5041 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     5101 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     5161 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     5221 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     5281 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     5341 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     5401 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     5461 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     5521 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     5581 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     5641 cagcagagat ccagtttggt tagtaccggg cccgctctag ccctgcagga actgtttaaa
     5701 ctgtgagggc ctatttccca tgattccttc atatttgcat atacgataca aggctgttag
     5761 agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg
     5821 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
     5881 tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa
     5941 aggacgaaac accgtgctcg ccaatggccg gctgtttaag agctatgctg gaaacagcat
     6001 agcaagttta aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     6061 tttttgtttg gcgcgcctat ggcttaatta acatggcggc cgcattctac cgggtagggg
     6121 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc
     6181 gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg
     6241 ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc
     6301 cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta
     6361 gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg
     6421 gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg
     6481 ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc
     6541 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc
     6601 cgggacccgg gatgcccaag aagaagagga aggtgtccaa tttactgacc gtacaccaaa
     6661 atttgcctgc attaccggtc gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca
     6721 tgttcaggga tcgccaggcg ttttctgagc atacctggaa aatgcttctg tccgtttgcc
     6781 ggtcgtgggc ggcatggtgc aagttgaata accggaaatg gtttcccgca gaacctgaag
     6841 atgttcgcga ttatcttcta tatcttcagg cgcgcggtct ggcagtaaaa actatccagc
     6901 aacatttggg ccagctaaac atgcttcatc gtcggtccgg gctgccacga ccaagtgaca
     6961 gcaatgctgt ttcactggtt atgcggcgga tccgaaaaga aaacgttgat gccggtgaac
     7021 gtgcaaaaca ggctctagcg ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg
     7081 aaaatagcga tcgctgccag gatatacgta atctggcatt tctggggatt gcttataaca
     7141 ccctgttacg tatagccgaa attgccagga tcagggttaa agatatctca cgtactgacg
     7201 gtgggagaat gttaatccat attggcagaa cgaaaacgct ggttagcacc gcaggtgtag
     7261 agaaggcact tagcctgggg gtaactaaac tggtcgagcg atggatttcc gtctctggtg
     7321 tagctgatga tccgaataac tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc
     7381 catctgccac cagccagcta tcaactcgcg ccctggaagg gatttttgaa gcaactcatc
     7441 gattgattta cggcgctaag gatgactctg gtcagagata cctggcctgg tctggacaca
     7501 gtgcccgtgt cggagccgcg cgagatatgg cccgcgctgg agtttcaata ccggagatca
     7561 tgcaagctgg tggctggacc aatgtaaata ttgtcatgaa ctatatccgt aacctggata
     7621 gtgaaacagg ggcaatggtg cgcctgctgg aagatggcga ttagccatgt ttaagggttc
     7681 cggttccact aggtacaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
     7741 atttgtgaaa gattgactgg tattcttaa
//
