LOCUS       sequence-240462-        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     91..110
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(209..655)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDWAMSG
                     LGRSRCRLDLVRLDCRLDGSSPRPLIAQSNSRGRHGGREHATSGPIRPIVSRITIHWPS
                     FYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     323..345
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     337..354
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     338..354
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     361..380
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        361..379
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      410..419
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      414..423
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(615..633)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(616..633)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(651..667)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(651..667)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(664..686)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    675..691
                     /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(699..705,706..723,724..729))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    744..765
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(882..899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1053..1641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1133..1152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1812..2603,2604..2672))
                     /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     2435..2454
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2673..2777)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(2855..3113,1..197))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2992..3013)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
       61 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
      121 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
      181 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
      241 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
      301 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg
      361 taatacgact cactataggg cgaattgggc ccgacgtcgc atgctcccgg ccgccatggc
      421 ggccgcggga attcgattgg gcgatgagtg gtctcgggga ggatccgtct agacggcagt
      481 ctagacgtac taagtctaga cggcatctag acctcccgag accactcatc gcccaatcac
      541 tagtgaattc gcggccgcct gcaggtcgac catatgggag agctcccaac gcgttggatg
      601 catagcttga gtattctata gtgtcaccta aatagcttgg cgtaatcatg gtcatagctg
      661 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
      721 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
      781 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
      841 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
      901 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      961 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1021 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1081 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1141 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1201 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1261 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1321 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1381 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1441 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1501 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1561 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1621 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1681 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1741 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1801 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1861 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1921 ccatctggcc ccagtgctgc aatgataccg cgcgacccac gctcaccggc tccagattta
     1981 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2041 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2101 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2161 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2221 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2281 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2341 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2401 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2461 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2521 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2581 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2641 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2701 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2761 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgatgcggt gtgaaatacc
     2821 gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta
     2881 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2941 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3001 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3061 cagggcgctg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtc
//
