LOCUS       sequence-352718-        8250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses  Cebpa_v1 (mU6), Cebpb_v1 (hU6) gRNAs and Cre-recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8250)
  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 8250)
  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: mU6-sgCebpa_v1-hu6-sgCebpb_v1
FEATURES             Location/Qualifiers
     source          1..8250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             225..405
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(467..1055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(547..566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1226..2017,2018..2086))
                     /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     1849..1868
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2087..2191)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2266..2285)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2457..2836
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2838..3036
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2991..3011
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3054..3234
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3281..3406
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3903..4136
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4321..4365
                     /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    4663..4780
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     5093..5113
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     5339..5359
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5510..5529
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        5750..6249
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5785..5808)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6182..6201
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6253..6273
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6274..7302
                     /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    7358..7946
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7411..7431)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7829..7840)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7949..7965)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7950..7966)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
       61 agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga
      121 gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc
      181 tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag
      241 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
      301 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
      361 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag
      421 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
      481 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      541 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      601 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      661 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
      721 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
      781 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      841 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      901 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      961 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1021 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1081 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     1141 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1201 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1261 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1321 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     1381 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     1441 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     1501 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     1561 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     1621 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     1681 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     1741 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     1801 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     1861 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     1921 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     1981 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2041 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     2101 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     2161 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     2221 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     2281 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     2341 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     2401 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     2461 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     2521 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     2581 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     2641 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2701 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2761 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2821 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     2881 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     2941 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     3001 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
     3061 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     3121 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     3181 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     3241 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     3301 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     3361 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     3421 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     3481 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     3541 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     3601 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     3661 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     3721 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     3781 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     3841 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     3901 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     3961 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     4021 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     4081 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     4141 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     4201 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     4261 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     4321 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     4381 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     4441 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     4501 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     4561 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     4621 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     4681 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     4741 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     4801 cagagatcca gtttggttag taccgggccc gctctagccc tgcaggaact gtttaaactg
     4861 tgatccgacg cgccatctct aggcccgcgc cggccccctc gcacggactt gtgggagaag
     4921 ctcggctact cccctgcccc ggttaatttg catataatat ttcctagtaa ctatagaggc
     4981 ttaatgtgcg ataaaagaca gataatctgt tctttttaat actagctaca ttttacatga
     5041 taggcttgga tttctataag agatacaaat actaaattat tattttaaaa aacagcacaa
     5101 aaggaaactc accctaactg taaagtaatt gtgtgttttg agactataag tatcccttgg
     5161 agaaccacct tgttggtgct cgcagatgcc gcccaggttt gagagctaag cagaaagctg
     5221 catagcaagt tcaaataagg ctagtccgta cacaacttga aaaagtggca gccgagtcgg
     5281 ctgctttttt tgctcgaatc tacactcagc tatggcgcgc cccaaggtcg ggcaggaaga
     5341 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     5401 aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     5461 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
     5521 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaagccag
     5581 aaacatggat gggtctaaag gcggcggggt ttcagagcta agcacaagag tgcatagcaa
     5641 gttgaaataa ggctagtccg tttacaactt gaaaaagtgg cacccgagtc gggtgctttt
     5701 tttgtttggc gcgcctatgg cttaattaac atggcggccg cattctaccg ggtaggggag
     5761 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
     5821 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
     5881 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
     5941 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
     6001 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
     6061 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
     6121 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
     6181 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     6241 ggacccggga tgcccaagaa gaagaggaag gtgtccaatt tactgaccgt acaccaaaat
     6301 ttgcctgcat taccggtcga tgcaacgagt gatgaggttc gcaagaacct gatggacatg
     6361 ttcagggatc gccaggcgtt ttctgagcat acctggaaaa tgcttctgtc cgtttgccgg
     6421 tcgtgggcgg catggtgcaa gttgaataac cggaaatggt ttcccgcaga acctgaagat
     6481 gttcgcgatt atcttctata tcttcaggcg cgcggtctgg cagtaaaaac tatccagcaa
     6541 catttgggcc agctaaacat gcttcatcgt cggtccgggc tgccacgacc aagtgacagc
     6601 aatgctgttt cactggttat gcggcggatc cgaaaagaaa acgttgatgc cggtgaacgt
     6661 gcaaaacagg ctctagcgtt cgaacgcact gatttcgacc aggttcgttc actcatggaa
     6721 aatagcgatc gctgccagga tatacgtaat ctggcatttc tggggattgc ttataacacc
     6781 ctgttacgta tagccgaaat tgccaggatc agggttaaag atatctcacg tactgacggt
     6841 gggagaatgt taatccatat tggcagaacg aaaacgctgg ttagcaccgc aggtgtagag
     6901 aaggcactta gcctgggggt aactaaactg gtcgagcgat ggatttccgt ctctggtgta
     6961 gctgatgatc cgaataacta cctgttttgc cgggtcagaa aaaatggtgt tgccgcgcca
     7021 tctgccacca gccagctatc aactcgcgcc ctggaaggga tttttgaagc aactcatcga
     7081 ttgatttacg gcgctaagga tgactctggt cagagatacc tggcctggtc tggacacagt
     7141 gcccgtgtcg gagccgcgcg agatatggcc cgcgctggag tttcaatacc ggagatcatg
     7201 caagctggtg gctggaccaa tgtaaatatt gtcatgaact atatccgtaa cctggatagt
     7261 gaaacagggg caatggtgcg cctgctggaa gatggcgatt agccatgttt aagggttccg
     7321 gttccactag gtacaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     7381 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     7441 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     7501 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     7561 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     7621 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     7681 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     7741 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     7801 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     7861 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     7921 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgatcga gacctagaaa
     7981 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
     8041 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
     8101 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     8161 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
     8221 gctacttccc tgattggcag aactacacac
//
