LOCUS       sequence-456137-        8040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral or overpexression of cDNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8040)
  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 8040)
  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: HA-MOB1B
FEATURES             Location/Qualifiers
     source          1..8040
                     /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"
     misc_feature    1723..1840
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1909..2120
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2149..2175
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2149..2175
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2179..2205
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    join(2872..3446,3447..3449,3450..3458)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3039..3056)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3266..3285
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    3897..4485
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3950..3970)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4368..4379)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4557..4790
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4868..4989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4905..4924)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4959..4978
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5029..5164
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5184..5203)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5185..5203)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5213..5230)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5213..5229)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5222..5244)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5371..5826
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5458..5477)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5668..5689
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5852..5956
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5957..6025,6026..6817)
                     /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(6175..6194)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6988..7576
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7477..7496
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7730..7747
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7864..7885
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7900..7905,7906..7923,7924..7930)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7938..7954
                     /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     7943..7965
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7962..7978
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7962..7978
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7997..8017
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7999..8017
                     /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 cccagaccac aattttaaaa gaaaaggggg
     1741 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     1801 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     1861 cagagatcca ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg gcagagcgca
     1921 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     1981 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2041 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2101 ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg ccaccatgta cccctacgac
     2161 gtgcccgact acgccggcta tccgtatgat gtcccggact atgcaggaag cggaagcttc
     2221 ttgtttggta gtcgctcttc taaaactttt aaaccaaaga agaacattcc agagggttct
     2281 caccagtatg agctcttaaa acacgcagaa gccacacttg gcagtggcaa ccttcggatg
     2341 gctgtcatgc ttcctgaagg ggaagatctc aatgaatggg ttgcagttaa cactgtggat
     2401 ttcttcaatc agatcaacat gctttatgga actatcacag acttctgtac agaagagagt
     2461 tgtccagtga tgtcagctgg cccaaaatat gagtatcatt gggcagatgg aacgaacata
     2521 aagaaaccta ttaagtgctc tgcaccaaag tatattgatt acttgatgac ttgggttcag
     2581 gaccagttgg atgatgagac gttatttcca tcaaaaattg gtgtcccgtt cccaaagaat
     2641 ttcatgtctg tggcaaaaac tatactcaaa cgcctcttta gggtttatgc tcacatttat
     2701 catcagcatt ttgaccctgt gatccagctt caggaggaag cacatctaaa tacatctttc
     2761 aagcacttta ttttttttgt ccaggaattc aaccttattg atagaagaga acttgcacca
     2821 ctccaagaac tgattgaaaa actcacctca aaagacagat gagcaattcc gcccctctcc
     2881 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     2941 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     3001 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     3061 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     3121 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     3181 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     3241 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     3301 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     3361 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     3421 tttcctttga aaaacacgat gataatatgg ccacaaccat gaagaccttc aacatctctc
     3481 agcaggacct tgagctggtg gaagttgcca ctgagaaaat caccatgctc tatgaggaca
     3541 acaagcacca tgttggtgct gccatcagga ccaagacagg agaaatcatc tctgctgtcc
     3601 acattgaagc ctacattggc agggtcactg tgtgtgcaga ggccattgcc attgggtctg
     3661 cagtctccaa tgggcagaag gactttgaca ccattgtggc tgtgaggcac ccctacagtg
     3721 atgaggtgga caggtccatc agagtggtgt ccccctgtgg catgtgcagg gagctcatct
     3781 cagactatgc ccctgattgc tttgttctga ttgagatgaa tggcaagctg gtcaagacaa
     3841 ccattgagga gctgattcca ctgaaataca ccagaaacta aacgcgttaa gtcgacaatc
     3901 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     3961 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     4021 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     4081 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     4141 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     4201 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     4261 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     4321 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     4381 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     4441 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga ctttaagacc
     4501 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     4561 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     4621 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     4681 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     4741 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtacgtatag
     4801 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     4861 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4921 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4981 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta
     5041 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5101 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5161 ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     5221 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     5281 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     5341 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     5401 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5461 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5521 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5581 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5641 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5701 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5761 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     5821 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5881 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5941 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6001 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     6061 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     6121 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     6181 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6241 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     6301 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6361 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6421 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6481 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6541 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6601 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6661 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6721 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6781 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6841 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6901 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6961 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     7021 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7081 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     7141 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7201 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7261 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7321 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7381 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7441 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7501 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7561 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7621 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     7681 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     7741 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     7801 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     7861 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     7921 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     7981 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
//
