LOCUS       sequence-470854-        6114 bp    DNA     circular SYN 02-MAY-2025
DEFINITION  Express 8X TEAD in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6114)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6114)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGL4 [luc2P/8X TEAD/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..6114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(33..39,40..41,42..48)
                     /label=2xGTIIC
                     /bound_moiety="TEF-1 family (TEAD) proteins"
                     /note="dimerized GTIIC motif from the SV40 enhancer, with 
                     high affinity for TEF-1 family (TEAD) proteins (Mahoney et 
                     al., 2005)"
     protein_bind    join(65..71,72..73,74..80)
                     /label=2xGTIIC
                     /bound_moiety="TEF-1 family (TEAD) proteins"
                     /note="dimerized GTIIC motif from the SV40 enhancer, with 
                     high affinity for TEF-1 family (TEAD) proteins (Mahoney et 
                     al., 2005)"
     protein_bind    join(119..125,126..127,128..134)
                     /label=2xGTIIC
                     /bound_moiety="TEF-1 family (TEAD) proteins"
                     /note="dimerized GTIIC motif from the SV40 enhancer, with 
                     high affinity for TEF-1 family (TEAD) proteins (Mahoney et 
                     al., 2005)"
     protein_bind    join(151..157,158..159,160..166)
                     /label=2xGTIIC
                     /bound_moiety="TEF-1 family (TEAD) proteins"
                     /note="dimerized GTIIC motif from the SV40 enhancer, with 
                     high affinity for TEF-1 family (TEAD) proteins (Mahoney et 
                     al., 2005)"
     promoter        181..212
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      239..248
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             245..1894
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             1898..2017
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2069..2190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2080..2099)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2134..2153
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2384..2741
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2384..2575
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2407..2427)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2592..2727
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2654..2673
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2772..3809
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3788..3796
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    3833..3881
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4037..4054)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4208..4796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4288..4307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4996..5787,5788..5856))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    5961..6009
                     /note="synthetic polyadenylation signal"
     misc_feature    6023..6114
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6063..6082
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gccattccac acattccaca ccttgcaaga
       61 gtcacattcc acacattcca ctgctaccaa ctagcccggc cagtgccaag ttgtgacaca
      121 ttccacacat tccacagcaa cgttgagaca cattccacac attccactaa gcttagacac
      181 tagagggtat ataatggaag ctcgacttcc agcttggcaa tccggtactg ttggtaaagc
      241 caccatggaa gatgccaaaa acattaagaa gggcccagcg ccattctacc cactcgaaga
      301 cgggaccgcc ggcgagcagc tgcacaaagc catgaagcgc tacgccctgg tgcccggcac
      361 catcgccttt accgacgcac atatcgaggt ggacattacc tacgccgagt acttcgagat
      421 gagcgttcgg ctggcagaag ctatgaagcg ctatgggctg aatacaaacc atcggatcgt
      481 ggtgtgcagc gagaatagct tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg
      541 tgtggctgtg gccccagcta acgacatcta caacgagcgc gagctgctga acagcatggg
      601 catcagccag cccaccgtcg tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt
      661 gcaaaagaag ctaccgatca tacaaaagat catcatcatg gatagcaaga ccgactacca
      721 gggcttccaa agcatgtaca ccttcgtgac ttcccatttg ccacccggct tcaacgagta
      781 cgacttcgtg cccgagagct tcgaccggga caaaaccatc gccctgatca tgaacagtag
      841 tggcagtacc ggattgccca agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt
      901 cagtcatgcc cgcgacccca tcttcggcaa ccagatcatc cccgacaccg ctatcctcag
      961 cgtggtgcca tttcaccacg gcttcggcat gttcaccacg ctgggctact tgatctgcgg
     1021 ctttcgggtc gtgctcatgt accgcttcga ggaggagcta ttcttgcgca gcttgcaaga
     1081 ctataagatt caatctgccc tgctggtgcc cacactattt agcttcttcg ctaagagcac
     1141 tctcatcgac aagtacgacc taagcaactt gcacgagatc gccagcggcg gggcgccgct
     1201 cagcaaggag gtaggtgagg ccgtggccaa acgcttccac ctaccaggca tccgccaggg
     1261 ctacggcctg acagaaacaa ccagcgccat tctgatcacc cccgaagggg acgacaagcc
     1321 tggcgcagta ggcaaggtgg tgcccttctt cgaggctaag gtggtggact tggacaccgg
     1381 taagacactg ggtgtgaacc agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag
     1441 cggctacgtt aacaaccccg aggctacaaa cgctctcatc gacaaggacg gctggctgca
     1501 cagcggcgac atcgcctact gggacgagga cgagcacttc ttcatcgtgg accggctgaa
     1561 gagcctgatc aaatacaagg gctaccaggt agccccagcc gaactggaga gcatcctgct
     1621 gcaacacccc aacatcttcg acgccggggt cgccggcctg cccgacgacg atgccggcga
     1681 gctgcccgcc gcagtcgtcg tgctggaaca cggtaaaacc atgaccgaga aggagatcgt
     1741 ggactatgtg gccagccagg ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt
     1801 ggacgaggtg cctaaaggac tgaccggcaa gttggacgcc cgcaagatcc gcgagattct
     1861 cattaaggcc aagaagggcg gcaagatcgc cgtgaattct cacggcttcc ctcccgaggt
     1921 ggaggagcag gccgccggca ccctgcccat gagctgcgcc caggagagcg gcatggatag
     1981 acaccctgct gcttgcgcca gcgccaggat caacgtctaa ggccgcgact ctagagtcgg
     2041 ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     2101 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2161 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2221 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     2281 aaaatcgata aggatccgtt tgcgtattgg gcgctcttcc gctgatctgc gcagcaccat
     2341 ggcctgaaat aacctctgaa agaggaactt ggttagctac cttctgaggc ggaaagaacc
     2401 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2461 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2521 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2581 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2641 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     2701 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcgat tcttctgaca
     2761 ctagcgccac catgaagaag cccgaactca ccgctaccag cgttgaaaaa tttctcatcg
     2821 agaagttcga cagtgtgagc gacctgatgc agttgtcgga gggcgaagag agccgagcct
     2881 tcagcttcga tgtcggcgga cgcggctatg tactgcgggt gaatagctgc gctgatggct
     2941 tctacaaaga ccgctacgtg taccgccact tcgccagcgc tgcactaccc atccccgaag
     3001 tgttggacat cggcgagttc agcgagagcc tgacatactg catcagtaga cgcgcccaag
     3061 gcgttactct ccaagacctc cccgaaacag agctgcctgc tgtgttacag cctgtcgccg
     3121 aagctatgga tgctattgcc gccgccgacc tcagtcaaac cagcggcttc ggcccattcg
     3181 ggccccaagg catcggccag tacacaacct ggcgggattt catttgcgcc attgctgatc
     3241 cccatgtcta ccactggcag accgtgatgg acgacaccgt gtccgccagc gtagctcaag
     3301 ccctggacga actgatgctg tgggccgaag actgtcccga ggtgcgccac ctcgtccatg
     3361 ccgacttcgg cagcaacaac gtcctgaccg acaacggccg catcaccgcc gtaatcgact
     3421 ggtccgaagc tatgttcggg gacagtcagt acgaggtggc caacatcttc ttctggcggc
     3481 cctggctggc ttgcatggag cagcagactc gctacttcga gcgccggcat cccgagctgg
     3541 ccggcagccc tcgtctgcga gcctacatgc tgcgcatcgg cctggatcag ctctaccaga
     3601 gcctcgtgga cggcaacttc gacgatgctg cctgggctca aggccgctgc gatgccatcg
     3661 tccgcagcgg ggccggcacc gtcggtcgca cacaaatcgc tcgccggagc gcagccgtat
     3721 ggaccgacgg ctgcgtcgag gtgctggccg acagcggcaa ccgccggccc agtacacgac
     3781 cgcgcgctaa ggaggtaggt cgagtttaaa ctctagaacc ggtcatggcc gcaataaaat
     3841 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gttcgaacta gatgctgtcg
     3901 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     3961 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     4021 agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4081 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4141 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4201 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4261 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4321 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4381 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4441 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4501 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4561 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4621 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4681 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4741 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4801 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4861 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4921 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagcggc cgcaaatgct
     4981 aaaccactgc agtggttacc agtgcttgat cagtgaggca ccgatctcag cgatctgcct
     5041 atttcgttcg tccatagtgg cctgactccc cgtcgtgtag atcactacga ttcgtgaggg
     5101 cttaccatca ggccccagcg cagcaatgat gccgcgagag ccgcgttcac cggcccccga
     5161 tttgtcagca atgaaccagc cagcagggag ggccgagcga agaagtggtc ctgctacttt
     5221 gtccgcctcc atccagtcta tgagctgctg tcgtgatgct agagtaagaa gttcgccagt
     5281 gagtagtttc cgaagagttg tggccattgc tactggcatc gtggtatcac gctcgtcgtt
     5341 cggtatggct tcgttcaact ctggttccca gcggtcaagc cgggtcacat gatcacccat
     5401 attatgaaga aatgcagtca gctccttagg gcctccgatc gttgtcagaa gtaagttggc
     5461 cgcggtgttg tcgctcatgg taatggcagc actacacaat tctcttaccg tcatgccatc
     5521 cgtaagatgc ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg agtagtgtat
     5581 acggcgacca agctgctctt gcccggcgtc tatacgggac aacaccgcgc cacatagcag
     5641 tactttgaaa gtgctcatca tcgggaatcg ttcttcgggg cggaaagact caaggatctt
     5701 gccgctattg agatccagtt cgatatagcc cactcttgca cccagttgat cttcagcatc
     5761 ttttactttc accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg ccgcaaagaa
     5821 gggaatgagt gcgacacgaa aatgttggat gctcatactc gtcctttttc aatattattg
     5881 aagcatttat cagggttact agtacgtctc tcaaggataa gtaagtaata ttaaggtacg
     5941 ggaggtattg gacaggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     6001 ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
     6061 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctct
//
