LOCUS       sequence-340276-        7833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of V5-tagged AMOTL2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7833)
  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 7833)
  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-AMOTL2
FEATURES             Location/Qualifiers
     source          1..7833
                     /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"
     protein_bind    complement(3381..3405)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3424..3441)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3430..3654
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3700..4128
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3787..3806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3997..4018
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4137..4157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4142..4471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4322..4457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4384..4403
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4538..5332
                     /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(4592..4611)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5202..5221
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5506..5627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5543..5562)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5597..5616
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5676..5692)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5676..5692)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5689..5711)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5700..5716
                     /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(5724..5730,5731..5748,5749..5754))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5769..5790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5907..5924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6078..6666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6158..6177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6837..7628,7629..7697))
                     /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     7460..7479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7698..7802)
                     /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 ggacactgga agactcctcg gggacagtcc tgcaccgcct
     1081 catccaggag cagctgcgct acggcaacct gactgagacg cgcacgctgc tagccatcca
     1141 gcagcaggcc ctgaggggtg gggctggaac tgggggtaca gggagccccc aggcctccct
     1201 ggagatcctg gccccagagg acagtcaggt gctgcagcag gccaccaggc aggagcccca
     1261 gggccaggag caccagggcg gtgagaacca cctggcagag aacaccctct accggctatg
     1321 cccacagccc agcaagggag aggagctgcc cacctatgag gaggccaaag cccactcgca
     1381 gtactatgcg gcccagcagg cagggacccg gccacatgcg ggggaccgag atccccgtgg
     1441 ggccccggga ggcagtcgga ggcaggacga ggccctgcgg gagctgaggc atgggcacgt
     1501 gcgctcgttg agtgaacggc tccttcagtt gtccctggag aggaacggcg cccgggcccc
     1561 cagccacatg agctcctccc acagcttccc acagctggcc cgcaaccagc agggcccccc
     1621 actgaggggc ccccctgctg agggcccaga gtcccgagga cccccacctc agtaccctca
     1681 tgttgtacta gctcatgaga ccaccactgc tgtcactgac ccacggtacc gtgcccgcgg
     1741 cagcccgcac ttccagcatg ctgaagtcag gatcctgcag gcccaggtgc ctcctgtgtt
     1801 cctccaacag cagcagcagt accagtacct gcagcaatct caggagcacc cccctccccc
     1861 acatccagct gctctcggcc atggccccct gagctccctc agtccacctg ctgtggaggg
     1921 gccagtgagt gcccaggcct cctcagccac ctcgggcagt gcccacctgg cccagatgga
     1981 ggccgtgctg agggagaatg ccaggctgca gagagacaat gagcggctgc agagggagct
     2041 ggagagctct gcggagaagg ctggccgcat tgagaagctg gaaagcgaaa tccagcggct
     2101 ctctgaggcc catgagagcc tgaccagagc ctcctccaag cgtgaggccc tggagaagac
     2161 catgcggaac aagatggaca gtgaaatgag gaggctgcaa gacttcaacc gggatcttag
     2221 agagagattg gaatctgcaa atcgccgcct ggcaagcaag acacaggagg cccaggccgg
     2281 cagtcaggac atggtggcca agctgcttgc tcagagctac gaacagcagc aggagcaaga
     2341 gaagctggag cgagagatgg cactgctgcg cggcgccatc gaggaccagc ggcggcgtgc
     2401 cgagctgctg gagcaggctc tgggcaatgc gcagggccgg gcagctcgag ccgaagagga
     2461 gctgcgcaag aagcaggcct atgtggagaa agtggagcgg ctgcagcagg cgctcgggca
     2521 gctgcaggca gcctgtgaga agcgggagca gctggagctg cgtctgcgga ctcgcctgga
     2581 gcaggaactc aaggccctgc gtgcacagca gagacaggca ggtgccccag gtggtagcag
     2641 tggcagtggt gggtctccag agctcagcgc cctgcgactg tcagaacaac tgcgagagaa
     2701 ggaggagcag atcctggcgc tggaggccga catgaccaag tgggagcaga agtatttgga
     2761 ggaacgtgcc atgaggcagt ttgccatgga tgcggctgcc acggctgctg ctcagcgtga
     2821 caccactctc atccgacatt ccccccagcc ctcacccagc agcagcttca atgagggtct
     2881 gctcactggt ggccacaggc atcaggagat ggaaagcagg ttaaaggtgc tccatgccca
     2941 gatcctggag aaggatgcag tgatcaaggt ccttcagcag cgctccagga gagaccctgg
     3001 caaggccatc cagggctccc tgcggcctgc caagtcggtg ccatctgttt tcgcggctgc
     3061 ggcagcagga acccagggct ggcaagggct ctcttctagt gagcgacaaa cagcagacgc
     3121 ccctgctcgg ctgactacag cagacagagc acccacagag gagccagtgg tcacagctcc
     3181 ccctgctgcc catgccaaac acgggagcag agatgggagc acccagactg acggcccccc
     3241 agacagcacc tccacctgcc tgccaccgga gcctgacagc cttctggggt gcagcagtag
     3301 ccagagagca gcctctctgg actctgtagc tacatccaga gtccaggact tgtcagacat
     3361 ggtggagata ctgatctgag acccagcttt cttgtacaaa gtggtgataa acccgctgat
     3421 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3481 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3541 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3601 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3661 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3721 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3781 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3841 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3901 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3961 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4021 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4081 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4141 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4201 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4261 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4321 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4381 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4441 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4501 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4561 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4621 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4681 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4741 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4801 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4861 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4921 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4981 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5041 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5101 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5161 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5221 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5281 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5341 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5401 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5461 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5521 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5581 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5641 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5701 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5761 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5821 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5881 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5941 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6001 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6061 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6121 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6181 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6241 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6301 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6361 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6421 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6481 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6541 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6601 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6661 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6721 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6781 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6841 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6901 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6961 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7021 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7081 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7141 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7201 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7261 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7321 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7381 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7441 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7501 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7561 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7621 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7681 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7741 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7801 cgcacatttc cccgaaaagt gccacctgac gtc
//
