LOCUS       sequence-240580-        6775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression for funtional studies of cell growth promotion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6775)
  AUTHORS   Gaffney CJ, Oka T, Mazack V, Hilman D, Gat U, Muramatsu T, Inazawa 
            J, Golden A, Carey DJ, Farooq A, Tromp G, Sudol M
  TITLE     Identification, basic characterization and evolutionary analysis of 
            differentially spliced mRNA isoforms of human YAP1 gene.
  JOURNAL   Gene. 2012 Nov 10;509(2):215-22. doi: 10.1016/j.gene.2012.08.025. 
            Epub 2012 Aug 24.
  PUBMED    22939869
REFERENCE   2  (bases 1 to 6775)
  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"; doi: 
            "10.1016/j.gene.2012.08.025"; journalName: "Gene"; date: 
            "2012-11-10- 10"; volume: "509"; issue: "2"; pages: "215-22"
COMMENT     Sequence Label: pcDNA3.1 (-) hYAP 1-1beta
FEATURES             Location/Qualifiers
     source          1..6775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2132..2149)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2138..2362
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2408..2836
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2495..2514)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2705..2726
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2845..2865)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2850..3179
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3030..3165
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3092..3111
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3246..4040
                     /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(3300..3319)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3910..3929
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4214..4335
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4251..4270)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4305..4324
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4384..4400)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4384..4400)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4397..4419)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4408..4424
                     /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(4432..4438,4439..4456,4457..4462))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4477..4498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4615..4632)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4786..5374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4866..4885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5545..6336,6337..6405))
                     /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     6168..6187
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6406..6510)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6585..6604)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacgggccc tctagactcg agcggccgca tggatcccgg gcagcagccg
      721 ccgcctcaac cggcccccca gggccaaggg cagccgcctt cgcagccccc gcaggggcag
      781 ggcccgccgt ccggacccgg gcaaccggca cccgcggcga cccaggcggc gccgcaggca
      841 ccccccgccg ggcatcagat cgtgcacgtc cgcggggact cggagaccga cctggaggcg
      901 ctcttcaacg ccgtcatgaa ccccaagacg gccaacgtgc cccagaccgt gcccatgagg
      961 ctccggaagc tgcccgactc cttcttcaag ccgccggagc ccaaatccca ctcccgacag
     1021 gccagtactg atgcaggcac tgcaggagcc ctgactccac agcatgttcg agctcattcc
     1081 tctccagctt ctctgcagtt gggagctgtt tctcctggga cactgacccc cactggagta
     1141 gtctctggcc cagcagctac acccacagct cagcatcttc gacagtcttc ttttgagata
     1201 cctgatgatg tacctctgcc agcaggttgg gagatggcaa agacatcttc tggtcagaga
     1261 tacttcttaa atcacatcga tcagacaaca acatggcagg accccaggaa ggccatgctg
     1321 tcccagatga acgtcacagc ccccaccagt ccaccagtgc agcagaatat gatgaactcg
     1381 gcttcagcca tgaaccagag aatcagtcag agtgctccag tgaaacagcc accacccctg
     1441 gctccccaga gcccacaggg aggcgtcatg ggtggcagca actccaacca gcagcaacag
     1501 atgcgactgc agcaactgca gatggagaag gagaggctgc ggctgaaaca gcaagaactg
     1561 cttcggcagg tgaggccaca ggagttagcc ctgcgtagcc agttaccaac actggagcag
     1621 gatggtggga ctcaaaatcc agtgtcttct cccgggatgt ctcaggaatt gagaacaatg
     1681 acgaccaata gctcagatcc tttccttaac agtggcacct atcactctcg agatgagagt
     1741 acagacagtg gactaagcat gagcagctac agtgtccctc gaaccccaga tgacttcctg
     1801 aacagtgtgg atgagatgga tacaggtgat actatcaacc aaagcaccct gccctcacag
     1861 cagaaccgtt tcccagacta ccttgaagcc attcctggga caaatgtgga ccttggaaca
     1921 ctggaaggag atggaatgaa catagaagga gaggagctga tgccaagtct gcaggaagct
     1981 ttgagttctg acatccttaa tgacatggag tctgttttgg ctgccaccaa gctagataaa
     2041 gaaagctttc ttacatggtt ataggaattc caccacactg gactagtgga tccgagctcg
     2101 gtaccaagct taagtttaaa ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2161 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2221 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2281 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2341 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     2401 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2461 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2521 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2581 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2641 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2701 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2761 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2821 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2881 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2941 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3001 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3061 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3121 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3181 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     3241 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3301 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3361 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3421 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3481 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3541 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3601 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3661 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3721 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     3781 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3841 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3901 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3961 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4021 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4081 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4141 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4201 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4261 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4321 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4381 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4441 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4501 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4561 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4621 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4681 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4741 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4801 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4861 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4921 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4981 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5041 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5101 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5161 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5221 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5281 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5341 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5401 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5461 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5521 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5581 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5641 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5701 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5761 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5821 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5881 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5941 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6001 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6061 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6121 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6181 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6241 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6301 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6361 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6421 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6481 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6541 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     6601 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     6661 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     6721 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtt
//
