LOCUS       sequence-456434-        7563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knockout of MST1 and MST2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7563)
  AUTHORS   Klingbeil O, Skopelitis D, Tonelli C, Yoshimoto T, Alpsoy A, 
            Panepinto MC, Minicozzi F, Merrill JR, Cafiero AM, Aggarwal D, Russo
            S, Ha T, Demerdash OE, Wee TL, Spector DL, Lyons SK, Tuveson DA, 
            Cifani P, Vakoc CR
  TITLE     MARK2/MARK3 kinases are catalytic co-dependencies of YAP/TAZ in 
            human cancer.
  JOURNAL   Cancer Discov. 2024 Jul 26. doi: 10.1158/2159-8290.CD-23-1529.
  PUBMED    39058094
REFERENCE   2  (bases 1 to 7563)
  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: "Cancer 
            Discov. 2024 Jul 26. doi: 10.1158/2159-8290.CD-23-1529."
COMMENT     Sequence Label: sgMST1/2-1
FEATURES             Location/Qualifiers
     source          1..7563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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"
     promoter        1717..1965
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1717..1737
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1888..1907
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2546..2663
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2732..2943
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    3420..4008
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3473..3493)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3891..3902)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4080..4313
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4391..4512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4428..4447)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4482..4501
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4552..4687
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4707..4726)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4708..4726)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4736..4753)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4736..4752)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4745..4767)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4894..5349
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4981..5000)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5191..5212
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5375..5479
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5480..5548,5549..6340)
                     /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(5698..5717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6511..7099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7000..7019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7253..7270
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7387..7408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7423..7428,7429..7446,7447..7453)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7461..7477
                     /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     7466..7488
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7485..7501
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7485..7501
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7520..7540
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7522..7540
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct
     1741 tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta
     1801 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     1861 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     1921 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgatac accgagatat
     1981 caaggcgttt cagagctatg cacaagagtg catagcaagt tgaaataagg ctagtccgtt
     2041 atcaactttg ctggaaacag caaagtggca ccgagtcggt gctttttttg cgtgaccgag
     2101 cttgtctgcc cgcgcagtcc actagacaga cgcgcagagg tcgggcgggc ctagggccaa
     2161 tttcccatgg ttcccttcat ttgcatatat gatgtaataa ggttatggag actattaaac
     2221 ttagccctaa tcaaactata tgatgataat gtgtgtggta caaaaggtca taacttatta
     2281 tatactttga aacttaaaaa agggttacag tttagtcacc ataactgtaa aattttttct
     2341 attcttagct ttatatagtt cttgagaggc catgtttatc gagtactcca taacaatcca
     2401 ggtttaagag ctatgctgga aacagcatag caagtttaaa taaggctagt ccgttatcaa
     2461 ctttgctgga aacagcaaag tggcaccgag tcggtgcttt ttttgaattc tagatcttga
     2521 gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg
     2581 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     2641 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc
     2701 gccggctcga gtggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     2761 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
     2821 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     2881 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     2941 caggtgtcgt gacgcgggat ccgccaccat gatgaagacc ttcaacatct ctcagcagga
     3001 ccttgagctg gtggaagttg ccactgagaa aatcaccatg ctctatgagg acaacaagca
     3061 ccatgttggt gctgccatca ggaccaagac aggagaaatc atctctgctg tccacattga
     3121 agcctacatt ggcagggtca ctgtgtgtgc agaggccatt gccattgggt ctgcagtctc
     3181 caatgggcag aaggactttg acaccattgt ggctgtgagg cacccctaca gtgatgaggt
     3241 ggacaggtcc atcagagtgg tgtccccctg tggcatgtgc agggagctca tctcagacta
     3301 tgcccctgat tgctttgttc tgattgagat gaatggcaag ctggtcaaga caaccattga
     3361 ggagctgatt ccactgaaat acaccagaaa ctaatgtaca agtagcgcgt taagtcgaca
     3421 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3481 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3541 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3601 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3661 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3721 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3781 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3841 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3901 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3961 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag
     4021 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     4081 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
     4141 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     4201 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     4261 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta
     4321 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     4381 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4441 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4501 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
     4561 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4621 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4681 ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     4741 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     4801 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     4861 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     4921 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4981 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     5041 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5101 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     5161 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5221 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     5281 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     5341 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5401 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5461 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5521 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5581 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5641 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5701 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5761 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5821 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5881 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5941 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6001 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6061 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6121 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6181 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6241 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6301 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6361 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6421 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6481 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6541 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6601 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6661 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6721 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6781 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6841 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6901 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6961 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7021 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7081 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7141 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     7201 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     7261 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     7321 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     7381 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     7441 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     7501 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
     7561 agc
//
