LOCUS       sequence-272188-        3439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  7 copies the SMAD DNA binding motif.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3439)
  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 3439)
  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:7xSMAD_RE:MiniP
FEATURES             Location/Qualifiers
     source          1..3439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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     6..28
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     25..41
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     25..41
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        59..77
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     59..76
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    180..228
                     /note="synthetic polyadenylation signal"
     misc_feature    242..333
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     282..301
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        494..525
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      595..604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(638..657)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(639..657)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(664..681)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(664..680)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(673..695)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      821..1276
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(908..927)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1118..1139
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1354..1458
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1459..1527,1528..2319)
                     /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(1677..1696)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2490..3078
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2979..2998
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3232..3249
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3366..3387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3402..3407,3408..3425,3426..3432)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
       61 ttaggtgaca ctatagaata ctcaagctat gcatccaacg cgttgggagc tctcccatat
      121 ggtcgacctg caggcggccg cgaattcact agtgattggg cgatgagtgg tctcgggaga
      181 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac
      241 taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc
      301 ccagtgcaag tgcaggtgcc agaacatttc tctcgctgtc aggaggtcta gactgccgtc
      361 tagacttaac agtacgtcta gactcggccg tctagactta gtacgtctag actcggtatg
      421 tctagactga cccgtctaga cggatcctcc ctcgaggata tcaaatctgg cctcggcggc
      481 caagcttaga cactagaggg tatataatgg aagctcgact tccagcttgg caatccggta
      541 ctgttggtaa agccaccaat gcgagaccac tcatcgccca atcgaattcc cgcggccgcc
      601 atggcggccg ggagcatgcg acgtcgggcc caattcgccc tatagtgagt cgtattacaa
      661 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
      721 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
      781 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca
      841 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
      901 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
      961 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1021 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc cagcgccctg atagacggtt
     1081 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     1141 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     1201 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     1261 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     1321 accgcatcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1381 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1441 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1501 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1561 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1621 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1681 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     1741 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1801 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1861 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1921 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1981 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2041 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2101 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2161 gccggtgagc gtgggtcgcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2221 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2281 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2341 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2401 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2461 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2521 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2581 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2641 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2701 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2761 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2821 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2881 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2941 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3001 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3061 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3121 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3181 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3241 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3301 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3361 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     3421 gctcgtatgt tgtgtggaa
//
