LOCUS       sequence-272282-        3426 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3 Copies of the Daf-16 family binding element.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3426)
  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 3426)
  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:3xDBE_RE:MiniP
FEATURES             Location/Qualifiers
     source          1..3426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..49
                     /note="synthetic polyadenylation signal"
     misc_feature    63..154
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     103..122
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        302..333
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      403..412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      407..416
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(446..465)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(447..465)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(472..489)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(472..488)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(481..503)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      629..1084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(716..735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     926..947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1162..1266
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1267..1335,1336..2127)
                     /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(1485..1504)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2298..2886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2787..2806
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3040..3057
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3174..3195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3210..3215,3216..3233,3234..3240)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3248..3264
                     /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)."
     primer_bind     3253..3275
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3272..3288
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3272..3288
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3306..3324
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3306..3323
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
       61 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      121 cccagtgcaa gtgcaggtgc cagaacattt ctctcgctga attcgatcaa gtaaacaact
      181 atgtaaacaa gatgcagatc aagtaaacaa ctatgtaaac aactccgttg atcaagtaaa
      241 caactatgta aacaatccct cgaggatatc aaatctggcc tcggcggcca agcttagaca
      301 ctagagggta tataatggaa gctcgacttc cagcttggca atccggtact gttggtaaag
      361 ccaccaatgc gagaccactc atcgcccaat cgaattcccg cggccgccat ggcggccggg
      421 agcatgcgac gtcgggccca attcgcccta tagtgagtcg tattacaatt cactggccgt
      481 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
      541 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
      601 acagttgcgc agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg
      661 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
      721 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
      781 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
      841 gattagggtg atggttcacg tagtgggcca gcgccctgat agacggtttt tcgccctttg
      901 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
      961 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     1021 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     1081 atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatcaggt
     1141 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1201 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1261 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1321 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1381 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1441 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1501 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1561 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1621 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1681 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1741 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1801 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1861 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1921 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1981 gggtcgcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     2041 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2101 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2161 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2221 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2281 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2341 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2401 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2461 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2521 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2581 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2641 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2701 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2761 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2821 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2881 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2941 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3001 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3061 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3121 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     3181 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     3241 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     3301 aagctattta ggtgacacta tagaatactc aagctatgca tccaacgcgt tgggagctct
     3361 cccatatggt cgacctgcag gcggccgcga attcactagt gattgggcga tgagtggtct
     3421 cgggag
//
