LOCUS       sequence-381605-        5676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid for split TEV assays; firefly luciferase reporter 
            gene driven by 10x clustered UAS binding sites linked to a minimal 
            MLP promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5676)
  AUTHORS   Wu Y, von Hauff IV, Jensen N, Rossner MJ, Wehr MC
  TITLE     Improved Split TEV GPCR beta-arrestin-2 Recruitment Assays via 
            Systematic Analysis of Signal Peptide and beta-arrestin Binding 
            Motif Variants.
  JOURNAL   Biosensors (Basel). 2022 Dec 29;13(1):48. doi: 10.3390/bios13010048.
  PUBMED    36671883
REFERENCE   2  (bases 1 to 5676)
  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"; journalName: "Biosensors 
            (Basel)."; date: "2022-12-29"; volume: "13(1):48. doi"; pages: " 
            10.3390/bios13010048"
COMMENT     Sequence Label: pGL4_10xUAS-MLPmin-luc2
FEATURES             Location/Qualifiers
     source          1..5676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      366..375
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             372..2024
                     /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"
     polyA_signal    2068..2189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2079..2098)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2133..2152
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2383..2740
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2383..2574
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2406..2426)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2591..2726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2653..2672
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2771..3370
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    3395..3443
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3599..3616)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3770..4358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3850..3869)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4558..5349,5350..5418))
                     /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    5523..5571
                     /note="synthetic polyadenylation signal"
     misc_feature    5585..5676
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5625..5644
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtaccg agtttctaga cggagtactg tcctccgagc ggagtactgt
       61 cctccgactc gagcggagta ctgtcctccg atcggagtac tgtcctccgc gaattccgga
      121 gtactgtcct ccgaagacgc tagtgccggt accgagtttc tagacggagt actgtcctcc
      181 gagcggagta ctgtcctccg actcgagcgg agtactgtcc tccgatcgga gtactgtcct
      241 ccgcgaattc cggagtactg tcctccgaag acgctagcgg ggggctataa aagggggtgg
      301 gggcgttcgt cctcactcta gatctgcgat ctaagtaagc ttggcaatcc ggtactgttg
      361 gtaaagccac catggaagat gccaaaaaca ttaagaaggg cccagcgcca ttctacccac
      421 tcgaagacgg gaccgccggc gagcagctgc acaaagccat gaagcgctac gccctggtgc
      481 ccggcaccat cgcctttacc gacgcacata tcgaggtgga cattacctac gccgagtact
      541 tcgagatgag cgttcggctg gcagaagcta tgaagcgcta tgggctgaat acaaaccatc
      601 ggatcgtggt gtgcagcgag aatagcttgc agttcttcat gcccgtgttg ggtgccctgt
      661 tcatcggtgt ggctgtggcc ccagctaacg acatctacaa cgagcgcgag ctgctgaaca
      721 gcatgggcat cagccagccc accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc
      781 tcaacgtgca aaagaagcta ccgatcatac aaaagatcat catcatggat agcaagaccg
      841 actaccaggg cttccaaagc atgtacacct tcgtgacttc ccatttgcca cccggcttca
      901 acgagtacga cttcgtgccc gagagcttcg accgggacaa aaccatcgcc ctgatcatga
      961 acagtagtgg cagtaccgga ttgcccaagg gcgtagccct accgcaccgc accgcttgtg
     1021 tccgattcag tcatgcccgc gaccccatct tcggcaacca gatcatcccc gacaccgcta
     1081 tcctcagcgt ggtgccattt caccacggct tcggcatgtt caccacgctg ggctacttga
     1141 tctgcggctt tcgggtcgtg ctcatgtacc gcttcgagga ggagctattc ttgcgcagct
     1201 tgcaagacta taagattcaa tctgccctgc tggtgcccac actatttagc ttcttcgcta
     1261 agagcactct catcgacaag tacgacctaa gcaacttgca cgagatcgcc agcggcgggg
     1321 cgccgctcag caaggaggta ggtgaggccg tggccaaacg cttccaccta ccaggcatcc
     1381 gccagggcta cggcctgaca gaaacaacca gcgccattct gatcaccccc gaaggggacg
     1441 acaagcctgg cgcagtaggc aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg
     1501 acaccggtaa gacactgggt gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga
     1561 tcatgagcgg ctacgttaac aaccccgagg ctacaaacgc tctcatcgac aaggacggct
     1621 ggctgcacag cggcgacatc gcctactggg acgaggacga gcacttcttc atcgtggacc
     1681 ggctgaagag cctgatcaaa tacaagggct accaggtagc cccagccgaa ctggagagca
     1741 tcctgctgca acaccccaac atcttcgacg ccggggtcgc cggcctgccc gacgacgatg
     1801 ccggcgagct gcccgccgca gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg
     1861 agatcgtgga ctatgtggcc agccaggtta caaccgccaa gaagctgcgc ggtggtgttg
     1921 tgttcgtgga cgaggtgcct aaaggactga ccggcaagtt ggacgcccgc aagatccgcg
     1981 agattctcat taaggccaag aagggcggca agatcgccgt gtaataattc tagagtcggg
     2041 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     2101 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2161 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2221 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     2281 aaatcgataa ggatccgttt gcgtattggg cgctcttccg ctgatctgcg cagcaccatg
     2341 gcctgaaata acctctgaaa gaggaacttg gttagctacc ttctgaggcg gaaagaacca
     2401 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2461 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2521 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2581 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2641 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     2701 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcgatt cttctgacac
     2761 tagcgccacc atgaccgagt acaagcctac cgtgcgcctg gccactcgcg atgatgtgcc
     2821 ccgcgccgtc cgcactctgg ccgccgcttt cgccgactac cccgctaccc ggcacaccgt
     2881 ggaccccgac cggcacatcg agcgtgtgac agagttgcag gagctgttcc tgacccgcgt
     2941 cgggctggac atcggcaagg tgtgggtagc cgacgacggc gcggccgtgg ccgtgtggac
     3001 tacccccgag agcgttgagg ccggcgccgt gttcgccgag atcggccccc gaatggccga
     3061 gctgagcggc agccgcctgg ccgcccagca gcaaatggag ggcctgcttg ccccccatcg
     3121 tcccaaggag cctgcctggt ttctggccac tgtaggagtg agccccgacc accagggcaa
     3181 gggcttgggc agcgccgtcg tgttgcccgg cgtagaggcc gccgaacgcg ccggtgtgcc
     3241 cgcctttctc gaaacaagcg caccaagaaa ccttccattc tacgagcgcc tgggcttcac
     3301 cgtgaccgcc gatgtcgagg tgcccgaggg acctaggacc tggtgtatga cacgaaaacc
     3361 tggcgcctaa tgatctagaa ccggtcatgg ccgcaataaa atatctttat tttcattaca
     3421 tctgtgtgtt ggttttttgt gtgttcgaac tagatgctgt cgaccgatgc ccttgagagc
     3481 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     3541 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct
     3601 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3661 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3721 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3781 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3841 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3901 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3961 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4021 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4081 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4141 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4201 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4261 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4321 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4381 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4441 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4501 gtatatatga gtaaacttgg tctgacagcg gccgcaaatg ctaaaccact gcagtggtta
     4561 ccagtgcttg atcagtgagg caccgatctc agcgatctgc ctatttcgtt cgtccatagt
     4621 ggcctgactc cccgtcgtgt agatcactac gattcgtgag ggcttaccat caggccccag
     4681 cgcagcaatg atgccgcgag agccgcgttc accggccccc gatttgtcag caatgaacca
     4741 gccagcaggg agggccgagc gaagaagtgg tcctgctact ttgtccgcct ccatccagtc
     4801 tatgagctgc tgtcgtgatg ctagagtaag aagttcgcca gtgagtagtt tccgaagagt
     4861 tgtggccatt gctactggca tcgtggtatc acgctcgtcg ttcggtatgg cttcgttcaa
     4921 ctctggttcc cagcggtcaa gccgggtcac atgatcaccc atattatgaa gaaatgcagt
     4981 cagctcctta gggcctccga tcgttgtcag aagtaagttg gccgcggtgt tgtcgctcat
     5041 ggtaatggca gcactacaca attctcttac cgtcatgcca tccgtaagat gcttttccgt
     5101 gaccggcgag tactcaacca agtcgttttg tgagtagtgt atacggcgac caagctgctc
     5161 ttgcccggcg tctatacggg acaacaccgc gccacatagc agtactttga aagtgctcat
     5221 catcgggaat cgttcttcgg ggcggaaaga ctcaaggatc ttgccgctat tgagatccag
     5281 ttcgatatag cccactcttg cacccagttg atcttcagca tcttttactt tcaccagcgt
     5341 ttcggggtgt gcaaaaacag gcaagcaaaa tgccgcaaag aagggaatga gtgcgacacg
     5401 aaaatgttgg atgctcatac tcgtcctttt tcaatattat tgaagcattt atcagggtta
     5461 ctagtacgtc tctcaaggat aagtaagtaa tattaaggta cgggaggtat tggacaggcc
     5521 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     5581 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
     5641 tccccagtgc aagtgcaggt gccagaacat ttctct
//
