LOCUS       sequence-456372-        8595 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 8595)
  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 8595)
  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-TAZ-4A
FEATURES             Location/Qualifiers
     source          1..8595
                     /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"
     regulatory      2140..2149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2152..2178
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2182..2208
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    join(3427..4001,4002..4004,4005..4013)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3594..3611)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3821..3840
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    4452..5040
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4505..4525)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4923..4934)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5112..5345
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5423..5544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5460..5479)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5514..5533
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5584..5719
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5739..5758)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5740..5758)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5768..5785)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5768..5784)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5777..5799)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5926..6381
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6013..6032)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6223..6244
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6407..6511
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6512..6580,6581..7372)
                     /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(6730..6749)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7543..8131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8032..8051
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8285..8302
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8419..8440
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8455..8460,8461..8478,8479..8485)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8493..8509
                     /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     8498..8520
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8517..8533
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8517..8533
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8552..8572
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8554..8572
                     /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 ccaccatggc ttacccctac
     2161 gacgttcccg actatgctgg ctacccctat gacgttcctg actacgcggg gagcatgaat
     2221 cccgcttcag cacctccgcc gctgccgccc ccaggtcagc aggtaataca tgtaacacag
     2281 gatctggaca cggacttgga agctttgttt aactccgtca tgaacccgaa gccatcaagc
     2341 tggagaaaga aaatattgcc tgaatctttc ttcaaagaac cagacgcggg tgcacacgca
     2401 cgacaatctt ccactgactc tagtggaggt catcctggac cccgcttggc gggaggcgca
     2461 cagcatgtaa gatcacatgc agcacctgcc agtctccagc ttggcacggg ggccggagct
     2521 gcggggtctc ctgcacaaca gcacgcccat ctgcggcagc aatcttatga tgtcactgat
     2581 gaacttccac tgcccccagg ctgggagatg acgtttaccg cgactgggca gaggtatttt
     2641 ctcaaccata tagagaagat aaccacttgg caagacccaa ggaaggccat gaaccagccc
     2701 ctcaatcaca tgaatctcca cccagctgtt tcttcaactc cagtcccgca gcgcagtatg
     2761 gcagtgagtc aaccgaatct ggttatgaat catcaacatc aacaacaaat ggctcccagt
     2821 acgctctcac aacaaaacca tccgacgcaa aatccccctg ctggactcat gtcaatgccc
     2881 aacgcgctta cgacccaaca acagcagcaa cagaagttga gactgcagag gatccaaatg
     2941 gagagggaaa gaatacggat gcgccaggag gaacttatga gacaagaagc tgcgctttgc
     3001 cgccagcttc ccatggaagc agagaccctg gccccggtac aagcggcggt gaatccccca
     3061 accatgacgc cggacatgag atctatcacc aataactcct ctgacccatt cttgaatgga
     3121 ggtccctacc atagccgaga acagtccacc gattctggtc tgggattggg atgctacagt
     3181 gtcccgacca ccccagaaga tttcctcagc aacgtcgacg aaatggatac tggtgaaaac
     3241 gctggtcaga cccctatgaa catcaaccca cagcagactc gatttccgga cttcctcgac
     3301 tgcttgccgg gcactaacgt cgacttggga acgctcgagt cagaggattt gataccgctc
     3361 ttcaacgacg tagaatccgc gctcaataaa agtgagccat tcctgacgtg gctctgagca
     3421 attccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
     3481 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
     3541 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
     3601 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
     3661 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
     3721 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     3781 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
     3841 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     3901 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
     3961 ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca accatgaaga
     4021 ccttcaacat ctctcagcag gaccttgagc tggtggaagt tgccactgag aaaatcacca
     4081 tgctctatga ggacaacaag caccatgttg gtgctgccat caggaccaag acaggagaaa
     4141 tcatctctgc tgtccacatt gaagcctaca ttggcagggt cactgtgtgt gcagaggcca
     4201 ttgccattgg gtctgcagtc tccaatgggc agaaggactt tgacaccatt gtggctgtga
     4261 ggcaccccta cagtgatgag gtggacaggt ccatcagagt ggtgtccccc tgtggcatgt
     4321 gcagggagct catctcagac tatgcccctg attgctttgt tctgattgag atgaatggca
     4381 agctggtcaa gacaaccatt gaggagctga ttccactgaa atacaccaga aactaaacgc
     4441 gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     4501 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     4561 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     4621 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     4681 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     4741 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     4801 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     4861 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     4921 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     4981 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     5041 gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     5101 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     5161 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     5221 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     5281 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     5341 tagcagtacg tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     5401 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     5461 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5521 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     5581 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5641 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5701 ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg
     5761 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     5821 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     5881 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     5941 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     6001 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     6061 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     6121 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     6181 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     6241 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     6301 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     6361 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6421 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6481 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6541 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6601 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6661 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6721 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6781 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6841 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6901 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6961 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     7021 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     7081 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     7141 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     7201 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     7261 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7321 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     7381 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7441 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7501 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7561 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7621 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7681 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7741 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7801 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7861 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7921 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7981 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     8041 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     8101 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     8161 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     8221 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     8281 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     8341 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     8401 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     8461 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     8521 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
     8581 gctggagctg caagc
//
