LOCUS       sequence-340280-        8745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of V5-tagged AMOT (p130).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8745)
  AUTHORS   Ma X, Mandausch FJ, Wu Y, Sahoo VK, Ma W, Leoni G, Hostiuc M, 
            Wintgens JP, Qiu J, Kannaiyan N, Rossner MJ, Wehr MC
  TITLE     Comprehensive split TEV based protein-protein interaction screening 
            reveals TAOK2 as a key modulator of Hippo signalling to limit 
            growth.
  JOURNAL   Cell Signal. 2024 Jan;113:110917. doi: 
            10.1016/j.cellsig.2023.110917. Epub 2023 Oct 7.
  PUBMED    37813295
REFERENCE   2  (bases 1 to 8745)
  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: "Cell 
            Signal."; date: "2024-01"; volume: "113:110917. doi"; pages: " 
            10.1016/j.cellsig.2023.110917. Epub 2023 Oct 7"
COMMENT     Sequence Label: pcDNA3.1_nV5-AMOT (p130)
FEATURES             Location/Qualifiers
     source          1..8745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             914..955
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             956..976
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    1007..1031
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2189..2209
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             3739..3756
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(4293..4317)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4342..4566
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4612..5040
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4699..4718)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4909..4930
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5049..5069)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5054..5383
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5234..5369
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5296..5315
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5450..6244
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5504..5523)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6114..6133
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6418..6539
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6455..6474)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6509..6528
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6588..6604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6588..6604)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6601..6623)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6612..6628
                     /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)."
     promoter        complement(join(6636..6642,6643..6660,6661..6666))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6681..6702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6819..6836)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6990..7578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7070..7089)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7749..8540,8541..8609))
                     /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     8372..8391
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8610..8714)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttacc atgggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacggaaaa
      961 cctgtatttt cagggcccgg atccgtcgac gaattctgca gatatcacaa gtttgtacaa
     1021 aaaagcaggc tccaccatga gaaattctga agaacagcca agtggaggga ccacggtatt
     1081 gcagcgtttg ctacaagagc agcttcgcta tggcaatcct agtgagaatc gcagcctgct
     1141 tgccatacac cagcaagcca cagggaatgg ccctcctttc cccagtggca gtgggaaccc
     1201 gggccctcag agtgatgtgt tgagtcccca agaccaccac caacagcttg tggctcatgc
     1261 tgctcgacaa gaaccccagg ggcaggaaat ccagtcagaa aacctcatca tggagaagca
     1321 gctgtctcct cgaatgcaaa ataatgaaga actcccgacc tatgaagaag ccaaggtcca
     1381 gtcccagtac tttcggggcc aacagcatgc cagtgttgga gctgccttct atgtcactgg
     1441 agtcaccaac cagaagatga ggactgaggg acgcccatca gttcagcggc tcaatcctgg
     1501 aaagatgcac caagatgagg gactcagaga ccttaagcaa gggcatgtcc gttccttgag
     1561 tgaacgacta atgcagatgt cactggccac cagtggagtt aaggcccatc cacctgttac
     1621 cagtgctccc ctctccccac cacaacccaa tgacctctac aagaatccca caagttccag
     1681 tgaattctac aaggcccaag ggccacttcc taaccagcat agcctgaagg gcatggaaca
     1741 ccgaggcccc ccaccagaat atcccttcaa gggcatgcca ccccaatctg tagtgtgcaa
     1801 gccccaagag ccagggcact tctatagtga gcatcgcctg aaccagccag ggagaacaga
     1861 ggggcaactg atgaggtatc agcatccccc tgagtatgga gcagccaggc cagcgcagga
     1921 catctcattg ccattgtcag ccaggaactc gcagcctcac agccctactt cttctctgac
     1981 ctctgggggg tccttgccct tgctacaatc tccaccatcc actagattgt ctcctgcccg
     2041 acaccccttg gtcccaaacc agggagacca ttcagctcac ctgcctaggc cgcagcagca
     2101 tttccttcct aatcaggctc accaggggga tcattaccgt ctctcccaac ctggcctgag
     2161 tcagcagcag cagcaacagc agcagcagca ccatcatcac catcaccacc aacaacagca
     2221 gcagcagcag ccacagcagc agccaggaga agcctattca gctatgcctc gggctcagcc
     2281 atcctctgct tcttatcagc cagtgccagc agaccctttt gccattgttt ccagagccca
     2341 gcagatggtt gagatcctct cagacgagaa ccggaacttg aggcaagagt tggaaggatg
     2401 ctatgagaag gtggcaagac tgcagaaggt ggagacagaa atccagcgcg tctcggaggc
     2461 atatgagaac ctcgtgaagt catcctccaa aagagaggcc ctagagaaag ccatgagaaa
     2521 caagctagag ggcgagattc ggaggatgca tgatttcaac agggatctga gagagcgtct
     2581 agagactgcc aacaagcagc ttgcagagaa ggaatatgag gggtcagagg acaccagaaa
     2641 aaccatctcg cagctctttg caaaaaataa agaaagccag cgtgagaagg agaagctgga
     2701 agcggagctg gccactgccc gttctaccaa tgaggaccaa agacgacaca tcgaaatccg
     2761 agatcaggcc ctgagtaatg cccaggccaa ggtggtaaag ctggaagaag agctgaaaaa
     2821 gaagcaagtg tacgttgaca aggtggagaa gatgcagcag gcccttgtac agctccaggc
     2881 agcatgtgaa aaacgtgagc agctagagca ccgtctccgg acacgactgg agagggaact
     2941 ggaatccctg agaatccagc agcgtcaggg caactgtcag cccaccaacg tttcagaata
     3001 caatgctgct gcactgatgg agctccttcg ggagaaagag gagaggattc tggctctgga
     3061 agctgatatg acaaagtggg agcagaaata tttggaggag aatgtgatga gacattttgc
     3121 tctggatgct gctgcaactg tggctgctca gagggacaca acagtcatca gtcactctcc
     3181 taacaccagc tatgacacag ctctagaagc tcgcatccag aaagaggagg aagaaatctt
     3241 gatggccaat aagcgttgcc ttgacatgga gggcaggatt aagaccctcc atgcccagat
     3301 tattgagaag gatgccatga tcaaagtact ccagcagcgt tcccggaagg agccgagcaa
     3361 gacagagcag ctgtcgtgca tgcggccagc gaagtctctg atgtccattt ccaatgctgg
     3421 atcaggcttg ctctcccact catccaccct gactggctcc cccatcatgg aagaaaagcg
     3481 agacgacaag agctggaagg ggagcctagg cattctcctg ggtggagact accgtgctga
     3541 atatgtccct tccacaccct cgcctgtgcc accctcgact cccctgctct cggctcactc
     3601 caagacagac agccgagact gcagtaccca aactgaacgt gggacggaat cgaacaaaac
     3661 tgcagctgtt gctcccatct ctgttcctgc tccagttgct gctgccgcca ctgctgccgc
     3721 catcactgcc actgctgcca ccatcaccac caccatggta gctgctgctc cagttgctgt
     3781 tgctgctgct gctgctccag ctgctgctgc tgccccgtct ccagccactg ccgctgctac
     3841 tgctgctgct gtttctccag ctgctgctgg tcagattcca gctgctgcct ctgttgcctc
     3901 agctgctgcc gttgctcctt ctgctgctgc tgctgctgct gttcaggttg ctccagctgc
     3961 tccggctcca gttccagctc cggctctggt tccggtccca gctccagcag cggctcaggc
     4021 ttctgctcct gctcagactc aggcaccaac ttcagctccg gctgtggctc caactccagc
     4081 tccaactcca actccagctg tggctcaggc tgaggttcct gcaagtccag ctaccggtcc
     4141 tggaccacat cgtttgtcta taccaagttt gacctgcaat ccagacaaaa cagatgggcc
     4201 tgtgttccac tccaatactc tggaaagaaa aactcccatt cagatcctgg gacaagagcc
     4261 tgatgcagag atggtggaat atctcatcta agacccagct ttcttgtaca aagtggtgat
     4321 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     4381 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     4441 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     4501 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     4561 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     4621 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4681 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4741 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4801 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4861 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4921 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4981 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5041 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     5101 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     5161 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     5221 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5281 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     5341 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     5401 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     5461 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5521 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5581 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     5641 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5701 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5761 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5821 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5881 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5941 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     6001 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     6061 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     6121 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     6181 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6241 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6301 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     6361 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     6421 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6481 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6541 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     6601 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6661 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6721 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6781 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6841 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6901 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6961 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7021 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7081 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7141 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7201 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7261 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7321 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7381 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     7441 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7501 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7561 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7621 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7681 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7741 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7801 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7861 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7921 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7981 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     8041 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     8101 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     8161 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     8221 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8281 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     8341 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     8401 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8461 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8521 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8581 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8641 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8701 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
