LOCUS       sequence-340451-        8361 bp    DNA     circular SYN 14-APR-2025
DEFINITION  constitutive expression of V5-tagged AMOTL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8361)
  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 8361)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-AMOTL1
FEATURES             Location/Qualifiers
     source          1..8361
                     /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(3909..3933)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3952..3969)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3958..4182
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4228..4656
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4315..4334)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4525..4546
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4665..4685)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4670..4999
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4850..4985
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4912..4931
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5066..5860
                     /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(5120..5139)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5730..5749
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6034..6155
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6071..6090)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6125..6144
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6204..6220)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6204..6220)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6217..6239)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6228..6244
                     /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(6252..6258,6259..6276,6277..6282))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6297..6318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6435..6452)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6606..7194)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6686..6705)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7365..8156,8157..8225))
                     /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     7988..8007
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8226..8330)
                     /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 tccaccatgt ggagggcaaa gttgcgccgg ggaacttgtg agcctgcggt
     1081 gaaagggtcc ccttctgctt gttatagccc cagtagtcct gtccaggttc tagaagactc
     1141 cacctacttt tccccagact ttcagctcta ttctgggagg catgaaacat ctgctttgac
     1201 ggtggaggca accagtagca tcagggaaaa agttgttgaa gatcctcttt gtaacttcca
     1261 ctccccaaac ttcctgagga tctcagaggt ggaaatgaga ggttccgagg atgcggcagc
     1321 tggaacagta ttgcagcggc tgatccagga acaactgcgg tatggcaccc caaccgagaa
     1381 catgaacttg ctggccattc agcaccaggc cacagggagt gcaggaccag cccatcctac
     1441 aaacaacttt tcttccacgg aaaacctcac tcaagaagac ccacaaatgg tctaccagtc
     1501 agcacgccaa gaaccgcagg gtcaagaaca ccaggtggac aatacggtga tggagaaaca
     1561 ggtccggtcc acgcagcctc agcagaacaa cgaggaactg cccacttacg aggaggccaa
     1621 agcacagtcg cagttcttca gggggcagca gcagcagcaa cagcagcagg gggcggtggg
     1681 ccatggttac tacatggcag ggggcaccag tcagaagtcc cgaactgagg ggaggcccac
     1741 tgtgaaccgt gccaacagtg gacaggcgca taaggacgag gcgctgaagg aactgaagca
     1801 gggccacgtc cgctcgctca gcgagagaat catgcagctg tccctggaga ggaatggggc
     1861 caagcaacac cttcccggct cggggaatgg aaagggcttc aaagtaggag gggggccctc
     1921 ccctgcccag cctgcaggta aagtgctgga ccctcggggt cctccacctg agtacccctt
     1981 caagaccaag caaatgatgt ccccagtcag caagacccag gagcacggac ttttttatgg
     2041 tgaccagcac cccgggatgc tccacgagat ggtcaagccc taccctgctc ctcagcctgt
     2101 gagaacagat gtggccgtcc tgcggtacca gccaccccct gagtatgggg taacgagccg
     2161 cccatgccaa cttccgttcc catcaaccat gcagcagcac agccccatgt cctcccagac
     2221 ctcttccgcc agcgggccac tgcactctgt ctccctgccg cttccactcc cgatggccct
     2281 gggtgctcca cagcccccgc ctgccgcctc ccccagccag cagcttggtc cagatgcctt
     2341 tgcgattgtg gagcgagccc agcaaatggt ggagatatta acagaggaga accgggtgct
     2401 tcaccaggaa cttcagggtt actacgacaa tgccgacaag ctccacaagt ttgaaaaaga
     2461 acttcagaga atttcggaag cctatgaaag tctggtcaag tctaccacca agcgagaatc
     2521 gctggacaag gccatgagaa acaaattgga aggcgagatt agaagacttc atgatttcaa
     2581 cagagacctc cgagatcgac tagagactgc taacaggcaa ctatccagca gggaatacga
     2641 agggcatgaa gacaaagctg cagaggggca ttatgcttcc cagaacaaag aattcttgaa
     2701 ggaaaaggag aaattagaaa tggagttagc agcagtgcgg actgcaagtg aggaccatcg
     2761 gagacacatc gagatcctgg accaggcttt gagcaacgcc caggccaggg tcatcaagct
     2821 ggaagaggag ttacgagaga agcaagcata tgttgagaaa gttgagaagc tgcagcaggc
     2881 cctgacccag ctgcagtctg catgtgagaa gcgagaacag atggagcgga gactgcggac
     2941 ttggctggag agagagctgg atgcactgag aacccagcag aaacatggaa atggccagcc
     3001 agccaacatg ccggaataca atgccccagc cctcctggaa cttgtgcggg agaaggagga
     3061 gcggatcctg gccctggagg ccgacatgac aaagtgggag cagaagtacc tggaggagag
     3121 caccatccga cactttgcca tgaatgccgc agccactgca gcagctgaga gggacaccac
     3181 gatcatcaac cactcacgga atggcagcta cggagagagc tcgctggagg cccacatctg
     3241 gcaagaggag gaggaggtgg tgcaggccaa cagaaggtgt caggacatgg aatacactat
     3301 taaaaatctc catgccaaaa tcatagagaa agatgctatg attaaggtcc tgcagcagcg
     3361 atctcgtaaa gatgccggga agacagactc ctccagccta cgtcctgccc gctccgttcc
     3421 atccatagca gcagctactg ggacacactc tcgccagacc tctcttacca gcagccagct
     3481 ggctgaggaa aagaaggaag agaagacctg gaaggggagc ataggattgc tgctggggaa
     3541 ggagcaccat gagcatgcct ctgccccact gctgccaccc ccacccacct cagcactgtc
     3601 ctccatagcc tccactacgg cagccagcag tgcccacgcc aagacaggca gcaaggacag
     3661 cagcacacag actgacaaga gtgccgagct cttctggccc agcatggcct cccttcccag
     3721 ccgcggccgg ctgagcacga cccctgctca cagccccgtc ctgaaacacc cagcggccaa
     3781 agggaccgca gagaaactgg agaactctcc tggccatggg aagtcgcctg accacagagg
     3841 ccgggtcagc agcttgctgc acaagcccga gttccctgat ggagagatga tggaagtcct
     3901 catctaagac ccagctttct tgtacaaagt ggtgataaac ccgctgatca gcctcgactg
     3961 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4021 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     4081 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4141 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     4201 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     4261 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4321 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4381 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4441 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4501 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4561 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4621 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     4681 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4741 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4801 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4861 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4921 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4981 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     5041 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     5101 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5161 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5221 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     5281 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5341 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5401 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5461 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5521 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5581 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5641 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5701 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5761 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5821 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5881 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5941 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     6001 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     6061 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6121 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     6181 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6241 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6301 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6361 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6421 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6481 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6541 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6601 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6661 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6721 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6781 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6841 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6901 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6961 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7021 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     7081 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7141 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7201 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7261 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     7321 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7381 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7441 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7501 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7561 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7621 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7681 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7741 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7801 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7861 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7921 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7981 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     8041 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     8101 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     8161 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     8221 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8281 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8341 cgaaaagtgc cacctgacgt c
//
