LOCUS       sequence-496940-        6205 bp    DNA     circular SYN 12-JAN-2026
DEFINITION  Dual-luciferase GLI reporter plasmid. 8xGLI binding site followed by
            minPro-luc2P expression, in addition to constitutive SV-40-driven 
            Renilla luciferase expression. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6205)
  AUTHORS   Krantz S, Bell B, Lund K, Parra NS, Ng Y, De Oliveira Rosa N, 
            Mukhopadhyay S, Croix BS, Sarin KY, Weigert R, Raimondi F, 
            Iglesias-Bartolome R
  TITLE     Dissection of Galphas and Hedgehog signaling crosstalk reveals 
            therapeutic opportunities to target adenosine receptor 2b in 
            Hedgehog-dependent tumors.
  JOURNAL   bioRxiv [Preprint]. 2025 Feb 27:2025.02.21.639530. doi: 
            10.1101/2025.02.21.639530.
  PUBMED    40060632
REFERENCE   2  (bases 1 to 6205)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 12, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Feb 27:2025.02.21.639530. doi: 
            10.1101/2025.02.21.639530."
COMMENT     Sequence Label: pGL4.R_8xGLIBS_minPro_Luc2p_Ren
FEATURES             Location/Qualifiers
     source          1..6205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    969..1017
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(1173..1190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1344..1932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1424..1443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2132..2923,2924..2992))
                     /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    3097..3145
                     /note="synthetic polyadenylation signal"
     misc_feature    3159..3250
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3199..3218
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        3624..3655
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      3682..3691
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3688..5337
                     /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             5341..5460
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    5512..5633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5523..5542)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5577..5596
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5827..6184
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5827..6018
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5850..5870)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6035..6170
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6097..6116
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 agcgccacca tgaccagcaa ggtgtacgac cccgagcaga ggaagaggat gatcaccggc
       61 ccccagtggt gggccaggtg caagcagatg aacgtgctgg acagcttcat caactactac
      121 gacagcgaga agcacgccga gaacgccgtg atcttcctgc acggcaacgc cgctagcagc
      181 tacctgtgga ggcacgtggt gccccacatc gagcccgtgg ccaggtgcat catccccgat
      241 ctgatcggca tgggcaagag cggcaagagc ggcaacggca gctacaggct gctggaccac
      301 tacaagtacc tgaccgcctg gttcgagctc ctgaacctgc ccaagaagat catcttcgtg
      361 ggccacgact ggggcgcctg cctggccttc cactacagct acgagcacca ggacaagatc
      421 aaggccatcg tgcacgccga gagcgtggtg gacgtgatcg agagctggga cgagtggcca
      481 gacatcgagg aggacatcgc cctgatcaag agcgaggagg gcgagaagat ggtgctggag
      541 aacaacttct tcgtggagac catgctgccc agcaagatca tgagaaagct ggagcccgag
      601 gagttcgccg cctacctgga gcccttcaag gagaagggcg aggtgagaag acccaccctg
      661 agctggccca gagagatccc cctggtgaag ggcggcaagc ccgacgtggt gcagatcgtg
      721 agaaactaca acgcctacct gagagccagc gacgacctgc ccaagatgtt catcgagagc
      781 gaccccggct tcttcagcaa cgccatcgtg gagggcgcca agaagttccc caacaccgag
      841 ttcgtgaagg tgaagggcct gcacttcagc caggaggacg cccccgacga gatgggcaag
      901 tacatcaaga gcttcgtgga gagagtgctg aagaacgagc agtaaactct agaaccggtc
      961 atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgttc
     1021 gaactagatg ctgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     1081 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     1141 taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1201 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1261 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1321 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1381 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1441 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1501 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1561 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1621 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1681 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1741 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1801 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1861 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1921 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1981 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2041 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2101 agcggccgca aatgctaaac cactgcagtg gttaccagtg cttgatcagt gaggcaccga
     2161 tctcagcgat ctgcctattt cgttcgtcca tagtggcctg actccccgtc gtgtagatca
     2221 ctacgattcg tgagggctta ccatcaggcc ccagcgcagc aatgatgccg cgagagccgc
     2281 gttcaccggc ccccgatttg tcagcaatga accagccagc agggagggcc gagcgaagaa
     2341 gtggtcctgc tactttgtcc gcctccatcc agtctatgag ctgctgtcgt gatgctagag
     2401 taagaagttc gccagtgagt agtttccgaa gagttgtggc cattgctact ggcatcgtgg
     2461 tatcacgctc gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg tcaagccggg
     2521 tcacatgatc acccatatta tgaagaaatg cagtcagctc cttagggcct ccgatcgttg
     2581 tcagaagtaa gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta cacaattctc
     2641 ttaccgtcat gccatccgta agatgctttt ccgtgaccgg cgagtactca accaagtcgt
     2701 tttgtgagta gtgtatacgg cgaccaagct gctcttgccc ggcgtctata cgggacaaca
     2761 ccgcgccaca tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct tcggggcgga
     2821 aagactcaag gatcttgccg ctattgagat ccagttcgat atagcccact cttgcaccca
     2881 gttgatcttc agcatctttt actttcacca gcgtttcggg gtgtgcaaaa acaggcaagc
     2941 aaaatgccgc aaagaaggga atgagtgcga cacgaaaatg ttggatgctc atactcgtcc
     3001 tttttcaata ttattgaagc atttatcagg gttactagta cgtctctcaa ggataagtaa
     3061 gtaatattaa ggtacgggag gtattggaca ggccgcaata aaatatcttt attttcatta
     3121 catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac
     3181 aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga
     3241 acatttctct ggcctaactg gccggtacac gcgtagattc gcgatcgacc tcgacaagca
     3301 gggaacaccc aagtagaagt tccgacaagc agggaacacc caagtagaag ctctcgacct
     3361 cgacaagcag ggaacaccca agtagaagct ccgacaagca gggaacaccc aagtagaagc
     3421 tctcgacctc gacaagcagg gaacacccaa gtagaagctc cgacaagcag ggaacaccca
     3481 agtagaagct ctcgacctcg acaagcaggg aacacccaag tagaagctcc gacaagcagg
     3541 gaacacccaa gtagaagctc tcgatcgcga atctgtggta ccgaggatat caagatctgg
     3601 cctcggcggc caagcttaga cactagaggg tatataatgg aagctcgact tccagcttgg
     3661 caatccggta ctgttggtaa agccaccatg gaagatgcca aaaacattaa gaagggccca
     3721 gcgccattct acccactcga agacgggacc gccggcgagc agctgcacaa agccatgaag
     3781 cgctacgccc tggtgcccgg caccatcgcc tttaccgacg cacatatcga ggtggacatt
     3841 acctacgccg agtacttcga gatgagcgtt cggctggcag aagctatgaa gcgctatggg
     3901 ctgaatacaa accatcggat cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc
     3961 gtgttgggtg ccctgttcat cggtgtggct gtggccccag ctaacgacat ctacaacgag
     4021 cgcgagctgc tgaacagcat gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa
     4081 gggctgcaaa agatcctcaa cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc
     4141 atggatagca agaccgacta ccagggcttc caaagcatgt acaccttcgt gacttcccat
     4201 ttgccacccg gcttcaacga gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc
     4261 atcgccctga tcatgaacag tagtggcagt accggattgc ccaagggcgt agccctaccg
     4321 caccgcaccg cttgtgtccg attcagtcat gcccgcgacc ccatcttcgg caaccagatc
     4381 atccccgaca ccgctatcct cagcgtggtg ccatttcacc acggcttcgg catgttcacc
     4441 acgctgggct acttgatctg cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag
     4501 ctattcttgc gcagcttgca agactataag attcaatctg ccctgctggt gcccacacta
     4561 tttagcttct tcgctaagag cactctcatc gacaagtacg acctaagcaa cttgcacgag
     4621 atcgccagcg gcggggcgcc gctcagcaag gaggtaggtg aggccgtggc caaacgcttc
     4681 cacctaccag gcatccgcca gggctacggc ctgacagaaa caaccagcgc cattctgatc
     4741 acccccgaag gggacgacaa gcctggcgca gtaggcaagg tggtgccctt cttcgaggct
     4801 aaggtggtgg acttggacac cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc
     4861 gtccgtggcc ccatgatcat gagcggctac gttaacaacc ccgaggctac aaacgctctc
     4921 atcgacaagg acggctggct gcacagcggc gacatcgcct actgggacga ggacgagcac
     4981 ttcttcatcg tggaccggct gaagagcctg atcaaataca agggctacca ggtagcccca
     5041 gccgaactgg agagcatcct gctgcaacac cccaacatct tcgacgccgg ggtcgccggc
     5101 ctgcccgacg acgatgccgg cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa
     5161 accatgaccg agaaggagat cgtggactat gtggccagcc aggttacaac cgccaagaag
     5221 ctgcgcggtg gtgttgtgtt cgtggacgag gtgcctaaag gactgaccgg caagttggac
     5281 gcccgcaaga tccgcgagat tctcattaag gccaagaagg gcggcaagat cgccgtgaat
     5341 tctcacggct tccctcccga ggtggaggag caggccgccg gcaccctgcc catgagctgc
     5401 gcccaggaga gcggcatgga tagacaccct gctgcttgcg ccagcgccag gatcaacgtc
     5461 taaggccgcg actctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
     5521 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     5581 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     5641 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     5701 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtttgcgtat tgggcgctct
     5761 tccgctgatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagc
     5821 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5881 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5941 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6001 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6061 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6121 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6181 caaaaagctc gattcttctg acact
//
