LOCUS       sequence-271994-        3394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  7 copies of the E-box motif.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3394)
  AUTHORS   Sarrion-Perdigones A, Gonzalez Y, Venken KJT
  TITLE     Rapid and Efficient Synthetic Assembly of Multiplex Luciferase 
            Reporter Plasmids for the Simultaneous Monitoring of Up to Six 
            Cellular Signaling Pathways.
  JOURNAL   Curr Protoc Mol Biol. 2020 Jun;131(1):e121. doi: 10.1002/cpmb.121.
  PUBMED    32539183
REFERENCE   2  (bases 1 to 3394)
  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: "Curr Protoc
            Mol Biol."; date: "2020-06"; volume: "131(1):e121. doi"; pages: " 
            10.1002/cpmb.121"
COMMENT     Sequence Label: TB:7xE-Box:MiniP
FEATURES             Location/Qualifiers
     source          1..3394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(79..870,871..939))
                     /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     702..721
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(940..1044)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1122..1577)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1259..1280)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1471..1490
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1703..1725
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1717..1734
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1718..1734
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1741..1760
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1741..1759
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1790..1799
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1794..1803
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1873..1904)
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    2020..2111
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(2052..2071)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    2125..2173
                     /note="synthetic polyadenylation signal"
     promoter        complement(2276..2294)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2277..2294)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2312..2328)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2312..2328)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2325..2347)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2336..2352
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2360..2366,2367..2384,2385..2390))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2405..2426
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2543..2560)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2714..3302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2794..2813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
       61 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
      121 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
      181 gggcttacca tctggcccca gtgctgcaat gataccgcgc gacccacgct caccggctcc
      241 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
      301 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
      361 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
      421 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
      481 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
      541 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
      601 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
      661 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
      721 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
      781 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
      841 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
      901 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
      961 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     1021 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg
     1081 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     1141 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1201 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     1261 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     1321 cgtctatcag ggcgctggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     1381 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     1441 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     1501 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     1561 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     1621 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     1681 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg
     1741 taatacgact cactataggg cgaattgggc ccgacgtcgc atgctcccgg ccgccatggc
     1801 ggccgcggga attcgattgg gcgatgagtg gtctcgcatt ggtggcttta ccaacagtac
     1861 cggattgcca agctggaagt cgagcttcca ttatataccc tctagtgtct aagcttggcc
     1921 gccgaggcca gatttgatat cctcgaggga cacgtgagca cgtgagcacg tgtccacgtg
     1981 cccgcacacg tgtccacgtg cacacgtgcg catgcagcga gagaaatgtt ctggcacctg
     2041 cacttgcact ggggacagcc tattttgcta gtttgttttg tttcgttttg ttttgatgga
     2101 gagcgtatgt tagtactatc gattcacaca aaaaaccaac acacagatgt aatgaaaata
     2161 aagatatttt attctcccga gaccactcat cgcccaatca ctagtgaatt cgcggccgcc
     2221 tgcaggtcga ccatatggga gagctcccaa cgcgttggat gcatagcttg agtattctat
     2281 agtgtcacct aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2341 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2401 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2461 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2521 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2581 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2641 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2701 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2761 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2821 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2881 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2941 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3001 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3061 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3121 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3181 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3241 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3301 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3361 aagggatttt ggtcatgaga ttatcaaaaa ggat
//
