LOCUS       sequence-417219-        7821 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus expressing shRNA to human LKB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7821)
  AUTHORS   Goodwin JM, Svensson RU, Lou HJ, Winslow MM, Turk BE, Shaw RJ
  TITLE     An AMPK-independent signaling pathway downstream of the LKB1 tumor 
            suppressor controls Snail1 and metastatic potential.
  JOURNAL   Mol Cell. 2014 Aug 7;55(3):436-50. doi: 
            10.1016/j.molcel.2014.06.021. Epub 2014 Jul 17.
  PUBMED    25042806
REFERENCE   2  (bases 1 to 7821)
  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"; doi: 
            "10.1016/j.molcel.2014.06.021"; journalName: "Mol Cell"; date: 
            "2014-08-7- 7"; volume: "55"; issue: "3"; pages: "436-50"
COMMENT     Sequence Label: pLentiX2-PURO-shLKB1-Hu
FEATURES             Location/Qualifiers
     source          1..7821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    228..249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(264..269,270..287,288..294)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    302..318
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     307..329
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     326..342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     326..342
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     361..381
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        363..381
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        409..635
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             636..816
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    863..988
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1481..1714
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1899..1943
                     /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    2210..2326
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2382..2970
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2435..2455)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2853..2864)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        2984..3483
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3019..3042)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3416..3435
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3504..4103
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3504..3523)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4000..4020
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    4284..4308
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    4441..4459
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(4505..4524)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    complement(4702..4726)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     LTR             4801..4981
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    5053..5174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5090..5109)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5144..5163
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5214..5349
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5369..5388)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5370..5388)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5398..5415)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5398..5414)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5407..5429)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5556..6011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5643..5662)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5853..5874
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6037..6141
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6142..6210,6211..7002)
                     /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     complement(6360..6379)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7173..7761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7662..7681
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
      361 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg
      421 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
      481 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
      541 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
      601 atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct
      661 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
      721 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
      781 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
      841 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
      901 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
      961 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     1021 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     1081 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     1141 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     1201 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     1261 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     1321 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     1381 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     1441 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     1501 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     1561 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     1621 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     1681 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     1741 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     1801 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     1861 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     1921 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     1981 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     2041 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     2101 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     2161 gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact tttaaaagaa
     2221 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     2281 tacaaactaa agaattacaa aaacaaatta caaaattcaa aattttatcg tccagtgtgg
     2341 tggaattctg cagatatcca gcacagtggc ggccgctcga caatcaacct ctggattaca
     2401 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2461 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2521 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     2581 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     2641 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     2701 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     2761 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     2821 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     2881 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     2941 gtcggatctc cctttgggcc gcctccccgc ctggaattct accgggtagg ggaggcgctt
     3001 ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca
     3061 agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc
     3121 tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc
     3181 ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg
     3241 cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag
     3301 cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg
     3361 ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc
     3421 tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt
     3481 tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     3541 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     3601 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     3661 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     3721 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     3781 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     3841 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     3901 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     3961 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     4021 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     4081 tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga
     4141 gcgcacgacc ccatgcatct cgagggcccg gtacctttaa gaccaatgac ttacaaggca
     4201 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct agactcgagc
     4261 ggccgccact gtgctggata tcaaccactt tgtacaagaa agctgggtct agatatctcg
     4321 agtgcggccg cgaattccac cacactggac tagtggatcc gagctcggta ccaagctttt
     4381 ccaaaaacga agagaagcag aaaatgggac agcacaccat tttctgcttc tcttcgggga
     4441 tctctatcac tgatagggaa cttataagat tcccaaatcc aaagacattt cacgtttatg
     4501 gtgatttccc agaacacata gcgacatgca aatattgcag ggcgccactc ccctgtccct
     4561 cacagccatc ttcctgccag ggcgcacgcg cgctgggtgt tcccgcctag tgacactggg
     4621 cccgcgattc cttggagcgg gttgatgacg tcagcgttcg aattcggtac cggatccagt
     4681 cgactgaatt ggttccttta aagcctgctt ttttgtacaa acttgttgat atctgcagaa
     4741 ttccaccaca ctggactagc tcactcccaa cgaagacaag atctgctttt tgcttgtact
     4801 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     4861 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     4921 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     4981 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     5041 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5101 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5161 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     5221 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5281 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5341 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     5401 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     5461 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     5521 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     5581 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5641 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5701 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5761 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5821 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5881 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     5941 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6001 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6061 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6121 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6181 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6241 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6301 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6361 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6421 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6481 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6541 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6601 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6661 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6721 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6781 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6841 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6901 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6961 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7021 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7081 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7141 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7201 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7261 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7321 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7381 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7441 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7501 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7561 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7621 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7681 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7741 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7801 tgctggcctt ttgctcacat g
//
