LOCUS       sequence-471813-        6076 bp    DNA     circular SYN 28-MAY-2025
DEFINITION  Express Gli response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6076)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6076)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGL4 [luc2P/Gli-RE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..6076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        143..174
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      201..210
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             207..1856
                     /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             1860..1979
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2031..2152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2042..2061)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2096..2115
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2346..2703
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2346..2537
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2369..2389)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2554..2689
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2616..2635
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2734..3771
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3750..3758
                     /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    3795..3843
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3999..4016)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4170..4758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4250..4269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4958..5749,5750..5818))
                     /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    5923..5971
                     /note="synthetic polyadenylation signal"
     misc_feature    5985..6076
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6025..6044
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgacc acccagacca cccagaccac ccagaccacc
       61 cagaccaccc agaccaccca gaccacccag accacccact cgaggatatc aagatctggc
      121 ctcggcggcc aagcttagac actagagggt atataatgga agctcgactt ccagcttggc
      181 aatccggtac tgttggtaaa gccaccatgg aagatgccaa aaacattaag aagggcccag
      241 cgccattcta cccactcgaa gacgggaccg ccggcgagca gctgcacaaa gccatgaagc
      301 gctacgccct ggtgcccggc accatcgcct ttaccgacgc acatatcgag gtggacatta
      361 cctacgccga gtacttcgag atgagcgttc ggctggcaga agctatgaag cgctatgggc
      421 tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg
      481 tgttgggtgc cctgttcatc ggtgtggctg tggccccagc taacgacatc tacaacgagc
      541 gcgagctgct gaacagcatg ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag
      601 ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat catacaaaag atcatcatca
      661 tggatagcaa gaccgactac cagggcttcc aaagcatgta caccttcgtg acttcccatt
      721 tgccacccgg cttcaacgag tacgacttcg tgcccgagag cttcgaccgg gacaaaacca
      781 tcgccctgat catgaacagt agtggcagta ccggattgcc caagggcgta gccctaccgc
      841 accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc catcttcggc aaccagatca
      901 tccccgacac cgctatcctc agcgtggtgc catttcacca cggcttcggc atgttcacca
      961 cgctgggcta cttgatctgc ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc
     1021 tattcttgcg cagcttgcaa gactataaga ttcaatctgc cctgctggtg cccacactat
     1081 ttagcttctt cgctaagagc actctcatcg acaagtacga cctaagcaac ttgcacgaga
     1141 tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc
     1201 acctaccagg catccgccag ggctacggcc tgacagaaac aaccagcgcc attctgatca
     1261 cccccgaagg ggacgacaag cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta
     1321 aggtggtgga cttggacacc ggtaagacac tgggtgtgaa ccagcgcggc gagctgtgcg
     1381 tccgtggccc catgatcatg agcggctacg ttaacaaccc cgaggctaca aacgctctca
     1441 tcgacaagga cggctggctg cacagcggcg acatcgccta ctgggacgag gacgagcact
     1501 tcttcatcgt ggaccggctg aagagcctga tcaaatacaa gggctaccag gtagccccag
     1561 ccgaactgga gagcatcctg ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc
     1621 tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa
     1681 ccatgaccga gaaggagatc gtggactatg tggccagcca ggttacaacc gccaagaagc
     1741 tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg actgaccggc aagttggacg
     1801 cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg cggcaagatc gccgtgaatt
     1861 ctcacggctt ccctcccgag gtggaggagc aggccgccgg caccctgccc atgagctgcg
     1921 cccaggagag cggcatggat agacaccctg ctgcttgcgc cagcgccagg atcaacgtct
     1981 aaggccgcga ctctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat
     2041 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2101 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2161 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     2221 gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg tttgcgtatt gggcgctctt
     2281 ccgctgatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttagct
     2341 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2401 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2461 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2521 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     2581 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     2641 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     2701 aaaaagctcg attcttctga cactagcgcc accatgaaga agcccgaact caccgctacc
     2761 agcgttgaaa aatttctcat cgagaagttc gacagtgtga gcgacctgat gcagttgtcg
     2821 gagggcgaag agagccgagc cttcagcttc gatgtcggcg gacgcggcta tgtactgcgg
     2881 gtgaatagct gcgctgatgg cttctacaaa gaccgctacg tgtaccgcca cttcgccagc
     2941 gctgcactac ccatccccga agtgttggac atcggcgagt tcagcgagag cctgacatac
     3001 tgcatcagta gacgcgccca aggcgttact ctccaagacc tccccgaaac agagctgcct
     3061 gctgtgttac agcctgtcgc cgaagctatg gatgctattg ccgccgccga cctcagtcaa
     3121 accagcggct tcggcccatt cgggccccaa ggcatcggcc agtacacaac ctggcgggat
     3181 ttcatttgcg ccattgctga tccccatgtc taccactggc agaccgtgat ggacgacacc
     3241 gtgtccgcca gcgtagctca agccctggac gaactgatgc tgtgggccga agactgtccc
     3301 gaggtgcgcc acctcgtcca tgccgacttc ggcagcaaca acgtcctgac cgacaacggc
     3361 cgcatcaccg ccgtaatcga ctggtccgaa gctatgttcg gggacagtca gtacgaggtg
     3421 gccaacatct tcttctggcg gccctggctg gcttgcatgg agcagcagac tcgctacttc
     3481 gagcgccggc atcccgagct ggccggcagc cctcgtctgc gagcctacat gctgcgcatc
     3541 ggcctggatc agctctacca gagcctcgtg gacggcaact tcgacgatgc tgcctgggct
     3601 caaggccgct gcgatgccat cgtccgcagc ggggccggca ccgtcggtcg cacacaaatc
     3661 gctcgccgga gcgcagccgt atggaccgac ggctgcgtcg aggtgctggc cgacagcggc
     3721 aaccgccggc ccagtacacg accgcgcgct aaggaggtag gtcgagttta aactctagaa
     3781 ccggtcatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     3841 gtgttcgaac tagatgctgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     3901 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     3961 aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc
     4021 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4081 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4141 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4201 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4261 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4321 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4381 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4441 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4501 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4561 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4621 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4681 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4741 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4801 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4861 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4921 tctgacagcg gccgcaaatg ctaaaccact gcagtggtta ccagtgcttg atcagtgagg
     4981 caccgatctc agcgatctgc ctatttcgtt cgtccatagt ggcctgactc cccgtcgtgt
     5041 agatcactac gattcgtgag ggcttaccat caggccccag cgcagcaatg atgccgcgag
     5101 agccgcgttc accggccccc gatttgtcag caatgaacca gccagcaggg agggccgagc
     5161 gaagaagtgg tcctgctact ttgtccgcct ccatccagtc tatgagctgc tgtcgtgatg
     5221 ctagagtaag aagttcgcca gtgagtagtt tccgaagagt tgtggccatt gctactggca
     5281 tcgtggtatc acgctcgtcg ttcggtatgg cttcgttcaa ctctggttcc cagcggtcaa
     5341 gccgggtcac atgatcaccc atattatgaa gaaatgcagt cagctcctta gggcctccga
     5401 tcgttgtcag aagtaagttg gccgcggtgt tgtcgctcat ggtaatggca gcactacaca
     5461 attctcttac cgtcatgcca tccgtaagat gcttttccgt gaccggcgag tactcaacca
     5521 agtcgttttg tgagtagtgt atacggcgac caagctgctc ttgcccggcg tctatacggg
     5581 acaacaccgc gccacatagc agtactttga aagtgctcat catcgggaat cgttcttcgg
     5641 ggcggaaaga ctcaaggatc ttgccgctat tgagatccag ttcgatatag cccactcttg
     5701 cacccagttg atcttcagca tcttttactt tcaccagcgt ttcggggtgt gcaaaaacag
     5761 gcaagcaaaa tgccgcaaag aagggaatga gtgcgacacg aaaatgttgg atgctcatac
     5821 tcgtcctttt tcaatattat tgaagcattt atcagggtta ctagtacgtc tctcaaggat
     5881 aagtaagtaa tattaaggta cgggaggtat tggacaggcc gcaataaaat atctttattt
     5941 tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag tactaacata cgctctccat
     6001 caaaacaaaa cgaaacaaaa caaactagca aaataggctg tccccagtgc aagtgcaggt
     6061 gccagaacat ttctct
//
