LOCUS       sequence-333563-        5064 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Luciferase reporter containing four copies of the Smad binding site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5064)
  AUTHORS   Zawel L, Dai JL, Buckhaults P, Zhou S, Kinzler KW, Vogelstein B, 
            Kern SE
  TITLE     Human Smad3 and Smad4 are sequence-specific transcription 
            activators.
  JOURNAL   Mol Cell. 1998 Mar . 1(4):611-7.
  PUBMED    9660945
REFERENCE   2  (bases 1 to 5064)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            1998 Mar . 1(4):611-7."
COMMENT     Sequence Label: SBE4-Luc
FEATURES             Location/Qualifiers
     source          1..5064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1555..1573
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      1636..1645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1675..1871)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1689..1824
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    1980..2071
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(2012..2031)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    2085..2133
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(2264..2719)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2401..2422)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2613..2632
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2746..2850
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2851..2919,2920..3711)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3069..3088)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3882..4470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4371..4390
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4624..4641
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4889..5010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4926..4945)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4980..4999
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5051..5064,1..1639))
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
ORIGIN
        1 ttccgccctt cttggccttt atgaggatct ctctgatttt tcttgcgtcg agttttccgg
       61 taagaccttt cggtacttcg tccacaaaca caactcctcc gcgcaacttt ttcgcggttg
      121 ttacttgact ggcgacgtaa tccacgatct ctttttccgt catcgtcttt ccgtgctcca
      181 aaacaacaac ggcggcggga agttcaccgg cgtcatcgtc gggaagacct gcgacacctg
      241 cgtcgaagat gttggggtgt tggagcaaga tggattccaa ttcagcggga gccacctgat
      301 agcctttgta cttaatcaga gacttcaggc ggtcaacgat gaagaagtgt tcgtcttcgt
      361 cccagtaagc tatgtctcca gaatgtagcc atccatcctt gtcaatcaag gcgttggtcg
      421 cttccggatt gtttacataa ccggacataa tcataggacc tctcacacac agttcgcctc
      481 tttgattaac gcccagcgtt ttcccggtat ccagatccac aaccttcgct tcaaaaaatg
      541 gaacaacttt accgaccgcg cccggtttat catccccctc gggtgtaatc agaatagctg
      601 atgtagtctc agtgagccca tatccttgcc tgatacctgg cagatggaac ctcttggcaa
      661 ccgcttcccc gacttcctta gagaggggag cgccaccaga agcaatttcg tgtaaattag
      721 ataaatcgta tttgtcaatc agagtgcttt tggcgaagaa ggagaatagg gttggcacca
      781 gcagcgcact ttgaatcttg taatcctgaa ggctcctcag aaacagctct tcttcaaatc
      841 tatacattaa gacgactcga aatccacata tcaaatatcc gagtgtagta aacattccaa
      901 aaccgtgatg gaatggaaca acacttaaaa tcgcagtatc cggaatgatt tgattgccaa
      961 aaataggatc tctggcatgc gagaatctca cgcaggcagt tctatgaggc agagcgacac
     1021 ctttaggcag accagtagat ccagaggagt tcatgatcag tgcaattgtc ttgtccctat
     1081 cgaaggactc tggcacaaaa tcgtattcat taaaaccggg aggtagatga gatgtgacga
     1141 acgtgtacat cgactgaaat ccctggtaat ccgttttaga atccatgata ataatttttt
     1201 ggatgattgg gagctttttt tgcacgttca aaattttttg caaccccttt ttggaaacga
     1261 acaccacggt aggctgcgaa atgcccatac tgttgagcaa ttcacgttca ttataaatgt
     1321 cgttcgcggg cgcaactgca actccgataa ataacgcgcc caacaccggc ataaagaatt
     1381 gaagagagtt ttcactgcat acgacgattc tgtgatttgt attcagccca tatcgtttca
     1441 tagcttctgc caaccgaacg gacatttcga agtactcagc gtaagtgatg tccacctcga
     1501 tatgtgcatc tgtaaaagca attgttccag gaaccagggc gtatctcttc atagccttat
     1561 gcagttgctc tccagcggtt ccatcttcca gcggatagaa tggcgccggg cctttcttta
     1621 tgtttttggc gtcttccatg gtggctttac caacagtacc ggaatgccaa gctttttgca
     1681 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     1741 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcga tggggcggag aatgggcgga
     1801 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     1861 attgagatgc agatcgcaga tctcgagccc gggctagcac gcgtaagagc tcggtacgtc
     1921 tagactgccg tctagactta gtacgtctag actgccgtct agacttagta cctatcgata
     1981 gagaaatgtt ctggcacctg cacttgcact ggggacagcc tattttgcta gtttgttttg
     2041 tttcgttttg ttttgatgga gagcgtatgt tagtactatc gattcacaca aaaaaccaac
     2101 acacagatgt aatgaaaata aagatatttt attgcggccg ctccaagtac ctcccgtacc
     2161 ttaatattac ttacttatca tggtagcttg ggctggcgta atagcgaaga ggcccgcacc
     2221 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcaaattgt aagcgttaat
     2281 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2341 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2401 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2461 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2521 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2581 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2641 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2701 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2761 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2821 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2881 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2941 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3001 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3061 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3121 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3181 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3241 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3301 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3361 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3421 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3481 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3541 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3601 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3661 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3721 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3781 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3841 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3901 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3961 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4021 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4081 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4141 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4201 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4261 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4321 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4381 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4441 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4501 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4561 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4621 cgcagcgagt cagtgagcga ggaagcggaa gagcgctgcc ggcacctgtc ctacgagttg
     4681 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     4741 ggagctgact gggttgaagg ctctcaaggg catcggtcga cggatcctta tcgattttac
     4801 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     4861 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     4921 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4981 tggtttgtcc aaactcatca atgtatctta tcatgtctgc tcgaagcggc cggccgcccc
     5041 gactctagaa ttacacggcg atct
//
