LOCUS       sequence-230543-        3113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GBPart - Operator/DNA Response element - 4 copies the SMAD DNA 
            binding motif.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3113)
  AUTHORS   Sarrion-Perdigones A, Chang L, Gonzalez Y, Gallego-Flores T, Young 
            DW, Venken KJT
  TITLE     Examining multiple cellular pathways at once using multiplex 
            hextuple luciferase assaying.
  JOURNAL   Nat Commun. 2019 Dec 13;10(1):5710. doi: 10.1038/s41467-019-13651-y.
  PUBMED    31836712
REFERENCE   2  (bases 1 to 3113)
  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: "Nat 
            Commun."; date: "2019-12-13"; volume: "10(1):5710. doi"; pages: " 
            10.1038/s41467-019-13651-y"
COMMENT     Sequence Label: p4xSMAD_RE
FEATURES             Location/Qualifiers
     source          1..3113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     138..157
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(256..702)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDWAMSG
                     LGRSRCRLDLVRLDCRLDGSSPRPLIAQSNSRGRHGGREHATSGPIRPIVSRITIHWPS
                     FYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     370..392
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     384..401
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     385..401
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     408..427
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        408..426
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      457..466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      461..470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(662..680)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(663..680)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(698..714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(698..714)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(711..733)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    722..738
                     /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(746..752,753..770,771..776))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    791..812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(929..946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1100..1688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1180..1199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1859..2650,2651..2719))
                     /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     2482..2501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2720..2824)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(2902..3113,1..244))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3039..3060)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gctggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
       61 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
      121 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
      181 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
      241 gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
      301 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      361 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac
      421 tatagggcga attgggcccg acgtcgcatg ctcccggccg ccatggcggc cgcgggaatt
      481 cgattgggcg atgagtggtc tcggggagga tccgtctaga cggcagtcta gacgtactaa
      541 gtctagacgg catctagacc tcccgagacc actcatcgcc caatcactag tgaattcgcg
      601 gccgcctgca ggtcgaccat atgggagagc tcccaacgcg ttggatgcat agcttgagta
      661 ttctatagtg tcacctaaat agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
      721 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
      781 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
      841 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
      901 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
      961 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1021 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1081 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1141 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1201 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1261 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1321 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1381 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1441 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1501 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     1561 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1621 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1681 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1741 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     1801 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     1861 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1921 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1981 gtgctgcaat gataccgcgc gacccacgct caccggctcc agatttatca gcaataaacc
     2041 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2101 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2161 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2221 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     2281 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     2341 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     2401 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2461 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     2521 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     2581 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     2641 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     2701 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     2761 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     2821 cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg aaataccgca cagatgcgta
     2881 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2941 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3001 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3061 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggc
//
