LOCUS       sequence-240581-        6811 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 6811)
  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 6811)
  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 1-1gamma
FEATURES             Location/Qualifiers
     source          1..6811
                     /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(2168..2185)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2174..2398
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2444..2872
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2531..2550)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2741..2762
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2881..2901)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2886..3215
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3066..3201
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3128..3147
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3282..4076
                     /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(3336..3355)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3946..3965
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4250..4371
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4287..4306)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4341..4360
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4420..4436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4420..4436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4433..4455)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4444..4460
                     /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(4468..4474,4475..4492,4493..4498))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4513..4534
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4651..4668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4822..5410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4902..4921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5581..6372,6373..6441))
                     /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     6204..6223
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6442..6546)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6621..6640)
                     /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 caatgcggaa tatcaatccc agcacagcaa attctccaaa atgtcaggag
     1621 ttagccctgc gtagccagtt accaacactg gagcaggatg gtgggactca aaatccagtg
     1681 tcttctcccg ggatgtctca ggaattgaga acaatgacga ccaatagctc agatcctttc
     1741 cttaacagtg gcacctatca ctctcgagat gagagtacag acagtggact aagcatgagc
     1801 agctacagtg tccctcgaac cccagatgac ttcctgaaca gtgtggatga gatggataca
     1861 ggtgatacta tcaaccaaag caccctgccc tcacagcaga accgtttccc agactacctt
     1921 gaagccattc ctgggacaaa tgtggacctt ggaacactgg aaggagatgg aatgaacata
     1981 gaaggagagg agctgatgcc aagtctgcag gaagctttga gttctgacat ccttaatgac
     2041 atggagtctg ttttggctgc caccaagcta gataaagaaa gctttcttac atggttatag
     2101 gaattccacc acactggact agtggatccg agctcggtac caagcttaag tttaaaccgc
     2161 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2221 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2281 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2341 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2401 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2461 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2521 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2581 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2641 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2701 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2761 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2821 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     2881 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2941 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3001 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3061 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3121 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3181 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3241 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3301 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3361 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3421 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3481 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3541 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3601 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3661 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3721 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3781 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     3841 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3901 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3961 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4021 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4081 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4141 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4201 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4261 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4321 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4381 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4441 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4501 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4561 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4621 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4681 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4741 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4801 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4861 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4921 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4981 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5041 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5101 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5161 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5221 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5281 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5341 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5401 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5461 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5521 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5581 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5641 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5701 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5761 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5821 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5881 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5941 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6001 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6061 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6121 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6181 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6241 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6301 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6361 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6421 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6481 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6541 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     6601 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     6661 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     6721 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     6781 cttcgcgatg tacgggccag atatacgcgt t
//
