LOCUS       Exported                6076 bp ds-DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6076)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pREBFB13 = pGL4[luc2P/Gli-RE/Hygro]
FEATURES             Location/Qualifiers
     source          1..6076
                     /organism="synthetic DNA construct"
                     /label=Firefly luciferase reporter vector
     misc_feature    complement(4..104)
                     /label=BFB874
                     /note="BFB874"
     misc_feature    27..98
                     /label=8x direct repeats of Gli consensus RE
                     /note="8x direct repeats of Gli consensus RE"
     promoter        144..174
                     /label=minimal promoter
                     /note="minimal promoter"
     gene            207..1982
                     /label=luc2
     CDS             207..1982
                     /codon_start=1
                     /label=firefly luciferase
                     /note="reporter gene; contains hPEST (P) destabilization 
                     sequence for rapid luciferase degradation"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAVNSHGFPPEVEEQAAGTLPMSCAQESG
                     MDRHPAACASARINV"
     polyA_signal    2022..2243
                     /label=SV40 late poly(A) signal
                     /note="SV40 late poly(A) signal"
     CDS             2734..3771
                     /codon_start=1
                     /label=synthetic hygromycin
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     primer_bind     3910..3929
                     /label=reporter vector primer 4 binding region
                     /note="reporter vector primer 4 binding region"
     rep_origin      4167
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="ColE1-derived plasmid replication origin"
     gene            complement(4958..5818)
                     /label=Amp(r)
     CDS             complement(4958..5818)
                     /codon_start=1
                     /label=synthetic beta-lactamase
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    5853..5887
                     /label=BFB861
                     /note="BFB861"
     polyA_signal    5923..6076
                     /label=poly
                     /note="poly(A) signal/transcriptional pause site"
     primer_bind     6025..6044
                     /label=RVprimer3 binding region
                     /note="RVprimer3 binding region"
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
//
