pT3-EF1aH Yap S127A

LOCUS       pT3-EF1aH Yap S127A               6693 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression of FLAG-tagged Yap containing S127A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    pT3-EF1aH Yap S127A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6693)
  AUTHORS   Tschaharganeh DF, Chen X, Latzko P, Malz M, Gaida MM, Felix K, Ladu 
            S, Singer S, Pinna F, Gretz N, Sticht C, Tomasi ML, Delogu S, Evert 
            M, Fan B, Ribback S, Jiang L, Brozzetti S, Bergmann F, Dombrowski F,
            Schirmacher P, Calvisi DF, Breuhahn K
  TITLE     Yes-Associated Protein Up-regulates Jagged-1 and Activates the NOTCH
            Pathway in Human Hepatocellular Carcinoma.
  JOURNAL   Gastroenterology. 2013 Jun;144(7):1530-1542.e12. doi: 
            10.1053/j.gastro.2013.02.009. Epub 2013 Feb 16.
  PUBMED    23419361
REFERENCE   2  (bases 1 to 6693)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6693
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1210..1234
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1265..1288
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(2800..2824)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2852..2871
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2906..2923)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2912..3136
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3554..3570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3554..3570)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3567..3589)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3578..3594
                     /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(3602..3632)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3647..3668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3785..3802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3956..4544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4036..4055)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4715..5575)
                     /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     5338..5357
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5576..5680)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5748..5766
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5804..5826)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5926..5945
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6139..6161
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6153..6170
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6154..6170
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6552..6572)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtaccaata
     1201 cggatatcaa caagtttgta caaaaaagca ggctttaaag gaaccaattc agtcgaccac
     1261 catggactac aaagacgatg acgataaagc aaggctcgaa tcggtaccta tggatcccgg
     1321 gcagcagccg ccgcctcaac cggcccccca gggccaaggg cagccgcctt cgcagccccc
     1381 gcaggggcag ggcccgccgt ccggacccgg gcaaccggca cccgcggcga cccaggcggc
     1441 gccgcaggca ccccccgccg ggcatcagat cgtgcacgtc cgcggggact cggagaccga
     1501 cctggaggcg ctcttcaacg ccgtcatgaa ccccaagacg gccaacgtgc cccagaccgt
     1561 gcccatgagg ctccggaagc tgcccgactc cttcttcaag ccgccggagc ccaaatccca
     1621 ctcccgacag gccagtactg atgcaggcac tgcaggagcc ctgactccac agcatgttcg
     1681 agctcatgcc tctccagctt ctctgcagtt gggagctgtt tctcctggga cactgacccc
     1741 cactggagta gtctctggcc cagcagctac acccacagct cagcatcttc gacagtcttc
     1801 ttttgagata cctgatgatg tacctctgcc agcaggttgg gagatggcaa agacatcttc
     1861 tggtcagaga tacttcttaa atcacatcga tcagacaaca acatggcagg accccaggaa
     1921 ggccatgctg tcccagatga acgtcacagc ccccaccagt ccaccagtgc agcagaatat
     1981 gatgaactcg gcttcaggtc ctcttcctga tggatgggaa caagccatga ctcaggatgg
     2041 agaaatttac tatataaacc ataagaacaa gaccacctct tggctagacc caaggcttga
     2101 ccctcgtttt gccatgaacc agagaatcag tcagagtgct ccagtgaaac agccaccacc
     2161 cctggctccc cagagcccac agggaggcgt catgggtggc agcaactcca accagcagca
     2221 acagatgcga ctgcagcaac tgcagatgga gaaggagagg ctgcggctga aacagcaaga
     2281 actgcttcgg caggagttag ccctgcgtag ccagttacca acactggagc aggatggtgg
     2341 gactcaaaat ccagtgtctt ctcccgggat gtctcaggaa ttgagaacaa tgacgaccaa
     2401 tagctcagat cctttcctta acagtggcac ctatcactct cgagatgaga gtacagacag
     2461 tggactaagc atgagcagct acagtgtccc tcgaacccca gatgacttcc tgaacagtgt
     2521 ggatgagatg gatacaggtg atactatcaa ccaaagcacc ctgccctcac agcagaaccg
     2581 tttcccagac taccttgaag ccattcctgg gacaaatgtg gaccttggaa cactggaagg
     2641 agatggaatg aacatagaag gagaggagct gatgccaagt ctgcaggaag ctttgagttc
     2701 tgacatcctt aatgacatgg agtctgtttt ggctgccacc aagctagata aagaaagctt
     2761 tcttacatgg ttataggcgg ccgcactcga gatatctaga cccagctttc ttgtacaaag
     2821 tggttgatat cgcggccgct gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2881 tatcatgtct ggatccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2941 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3001 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3061 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3121 gcggtgggct ctatggcttc tgaggcggga tccccttgaa atacatccac aggtacacct
     3181 ccaattgact caaatgatgt caattagtct atcagaagct tctaaagcca tgacatcatt
     3241 ttctggaatt ttccaagctg tttaaaggca cagtcaactt agtgtatgta aacttctgac
     3301 ccactggaat tgtgatacag tgaattataa gtgaaataat ctgtctgtaa acaattgttg
     3361 gaaaaatgac ttgtgtcatg cacaaagtag atgtcctaac tgacttgcca aaactattgt
     3421 ttgttaacaa gaaatttgtg gagtagttga aaaacgagtt ttaatgactc caacttaagt
     3481 gtatgtaaac ttccgacttc aactgtatag gtctagagtc gacctgcagg catgcaagct
     3541 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3601 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3661 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3721 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3781 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3841 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3901 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3961 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4021 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4081 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4141 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4201 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4261 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4321 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4381 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4441 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4501 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4561 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4621 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4681 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4741 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4801 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4861 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4921 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4981 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5041 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5101 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5161 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5221 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5281 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5341 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5401 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5461 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5521 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5581 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5641 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5701 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     5761 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     5821 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     5881 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     5941 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     6001 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     6061 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     6121 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc
     6181 tcggtaccct acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact
     6241 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta
     6301 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta
     6361 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt
     6421 tgactgtgcc tttaaacagc ttggaaaatt ccagaaaatg atgtcatggc tttagaagct
     6481 tctgatagac taattgacat catttgagtc aattggaggt gtacctgtgg atgtatttca
     6541 aggaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     6601 cagaagtatg caaagcatgc atatcgatac tagtttaatt aagatcctct ttgcagctaa
     6661 tggaccttct aggtcttgaa aggagtggga att
//