LOCUS       sequence-201226-        6265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  luciferase reporter with ARHGAP29 mutant promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6265)
  AUTHORS   Qiao Y, Chen J, Lim YB, Finch-Edmondson ML, Seshachalam VP, Qin L, 
            Jiang T, Low BC, Singh H, Lim CT, Sudol M
  TITLE     YAP Regulates Actin Dynamics through ARHGAP29 and Promotes 
            Metastasis.
  JOURNAL   Cell Rep. 2017 May 23;19(8):1495-1502. doi: 
            10.1016/j.celrep.2017.04.075.
  PUBMED    28538170
REFERENCE   2  (bases 1 to 6265)
  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.celrep.2017.04"; journalName: "Cell Rep"; date: 
            "2017-05-23- 23"; volume: "19"; issue: "8"; pages: "1495-1502"
COMMENT     Sequence Label: pGL3-BASIC- ARHGAP29 Mutant PROMOTER
FEATURES             Location/Qualifiers
     source          1..6265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    93..141
                     /note="synthetic polyadenylation signal"
     misc_feature    155..246
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     195..214
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     CDS             1482..1493
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      1782..1791
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1788..3440
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1854..1872)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3481..3602
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3492..3511)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3546..3565
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3850..3867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4021..4609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4101..4120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4780..5571,5572..5640))
                     /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     5403..5422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5641..5745)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5772..6227
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5859..5878)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6069..6090
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgatcggtgc gggcctcttc gctattacgc cagcccaagc taccatgata agtaagtaat
       61 attaaggtac gggaggtact tggagcggcc gcaataaaat atctttattt tcattacatc
      121 tgtgtgttgg ttttttgtgt gaatcgatag tactaacata cgctctccat caaaacaaaa
      181 cgaaacaaaa caaactagca aaataggctg tccccagtgc aagtgcaggt gccagaacat
      241 ttctctatcg ataggtaccg attgaatcgt agttgcactg ttcagcctcc ccttggcctg
      301 gggactcctt tgtctcctct tattagtttg cttcagttca ggcatcattg gcttccatgg
      361 tgagagtcct tggcgcattt ttctttttca gcacgattag aatttggttt ttgaggtttc
      421 ctatttgtgg caggaccttg ttactattat tttggggatt tgaggggaaa atctgtgtaa
      481 gttgagtcca aaaaatccag tatggtgcca aagtaaccaa ggagctgagt tttcttattt
      541 atcttttcta agtagcacct gaggagccaa atttgttgta ttttcttagc attgtttttc
      601 ctgtaagggt tcaaagatat gtctatgtaa gatgtccctc ttctctttct ttgtcaatga
      661 gtttttctct tgccccgcta ccctactttt ggttctgtca tgctcactta cttttctgtt
      721 tgttaagtgg gtagtctctc cttttatttt tcttccttat tttctctttt tgccttcact
      781 tatttctgcc ttccttggtt ttcttccctt ttcctgtgtc tgcacctcat tttctttcca
      841 gttttcctca gcacattctc ctgtgtcttt ttctgtttct cccgtgtgtg gtcatttttc
      901 tcgcagaccc cagcgctacc cacagtctcc ctccctgtgt tttttcctct gtttctctat
      961 gagttctttc tgaatgttcc agcactgagg taagtgccat cgcttgttcc agcgagtggt
     1021 gtgatatagg gactatgtgc tttctagccc tgcttgctat aattattcca tctttctcag
     1081 actttttttt ttcttctcat tatgaagtga gtttgaaagt gcagctaaga aagaaaggac
     1141 cgggcacagt ggctcatgct tgtaatccca gtactttggg aggctgaggc gggtggatca
     1201 cctgaggtcg ggaggtcgtc gagaccagct tggccaacat agcgaaaccc tgcctctact
     1261 aaaaatacta aaattagctt ggcgtggtaa tgggtgccta taatcgcagc tactcaggtg
     1321 gctgaggtgg gagaatcaat tgagcccggg aggcggaggt tgcagtgagc cgagatcacg
     1381 ccattagact cgtctgggcg acagagcgag actccgtctc cnaaaaaaaa aaaaaaaaaa
     1441 aaaaaantac agataggcct aagtggaaac tattaccctc catagagggc agaggggcag
     1501 agtagatggc tggttttgag gtagttcatt cttcattcag acgtgtggcg cccttcccca
     1561 ggccctaacc cctggaagag ggaggaaaaa ggcgatccac agaagtgtct cataagaaat
     1621 gtagctgctc atctggccat tagccgcatc tagtcctcag gtgacttttc tctgagtcgc
     1681 tctgcctgtc ccctgcaagc caaaagggca gccgacgcag cccgtgatgc tgggacagcg
     1741 tccggccgct gaaagcttgg cattccggta ctgttggtaa agccaccatg gaagacgcca
     1801 aaaacataaa gaaaggcccg gcgccattct atccgctgga agatggaacc gctggagagc
     1861 aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct tttacagatg
     1921 cacatatcga ggtggacatc acttacgctg agtacttcga aatgtccgtt cggttggcag
     1981 aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact
     2041 ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca gttgcgcccg
     2101 cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gggcatttcg cagcctaccg
     2161 tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa aagctcccaa
     2221 tcatccaaaa aattattatc atggattcta aaacggatta ccagggattt cagtcgatgt
     2281 acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt gtgccagagt
     2341 ccttcgatag ggacaagaca attgcactga tcatgaactc ctctggatct actggtctgc
     2401 ctaaaggtgt cgctctgcct catagaactg cctgcgtgag attctcgcat gccagagatc
     2461 ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt ccattccatc
     2521 acggttttgg aatgtttact acactcggat atttgatatg tggatttcga gtcgtcttaa
     2581 tgtatagatt tgaagaagag ctgtttctga ggagccttca ggattacaag attcaaagtg
     2641 cgctgctggt gccaacccta ttctccttct tcgccaaaag cactctgatt gacaaatacg
     2701 atttatctaa tttacacgaa attgcttctg gtggcgctcc cctctctaag gaagtcgggg
     2761 aagcggttgc caagaggttc catctgccag gtatcaggca aggatatggg ctcactgaga
     2821 ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg gtcggtaaag
     2881 ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg ctgggcgtta
     2941 atcaaagagg cgaactgtgt gtgagaggtc ctatgattat gtccggttat gtaaacaatc
     3001 cggaagcgac caacgccttg attgacaagg atggatggct acattctgga gacatagctt
     3061 actgggacga agacgaacac ttcttcatcg ttgaccgcct gaagtctctg attaagtaca
     3121 aaggctatca ggtggctccc gctgaattgg aatccatctt gctccaacac cccaacatct
     3181 tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc gccgccgttg
     3241 ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac gtcgccagtc
     3301 aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag
     3361 gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag gccaagaagg
     3421 gcggaaagat cgccgtgtaa ttctagagtc ggggcggccg gccgcttcga gcagacatga
     3481 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3541 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3601 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     3661 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg tcgaccgatg
     3721 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     3781 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     3841 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3901 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3961 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4021 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4081 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4141 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4201 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4261 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4321 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4381 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4441 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4501 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4561 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4621 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4681 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4741 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4801 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4861 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4921 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4981 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5041 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5101 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5161 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5221 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5281 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5341 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5401 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5461 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5521 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5581 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5641 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5701 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5761 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5821 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5881 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5941 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6001 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6061 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6121 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6181 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttgcc attcgccatt
     6241 caggctgcgc aactgttggg aaggg
//
