LOCUS       sequence-382578-        3189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription of 47xCAG and 12xBoxB repeats from a 
            tetracycline-inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3189)
  AUTHORS   Guo H, Ryan JC, Song X, Mallet A, Zhang M, Pabst V, Decrulle AL, 
            Ejsmont P, Wintermute EH, Lindner AB
  TITLE     Spatial engineering of E. coli with addressable phase-separated 
            RNAs.
  JOURNAL   Cell. 2022 Sep 29;185(20):3823-3837.e23. doi: 
            10.1016/j.cell.2022.09.016.
  PUBMED    36179672
REFERENCE   2  (bases 1 to 3189)
  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"; doi: 
            "10.1016/j.cell.2022.09"; journalName: "Cell"; date: "2022-09-29- 
            29"; volume: "185"; issue: "20"; pages: "3823-3837.e23"
COMMENT     Sequence Label: prCAG-BoxB
FEATURES             Location/Qualifiers
     source          1..3189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    183..201
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    227..245
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    251..269
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    295..313
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    319..337
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    363..381
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    409..427
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    453..471
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    477..495
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    521..539
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    545..563
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    589..607
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     terminator      635..706
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      722..749
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(777..794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(911..1456)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(991..1010)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1570..1664)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1685..2344)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2259..2278
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2345..2447)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(2477..3103)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     3008..3027
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        3119..3174
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    3155..3173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
       61 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
      121 cagcagcagc agcagcagca gagacggagc tcgtcgacgc ggccaagctt ggtaccgagc
      181 tcgggccctg aagaagggcc cctcgactaa gtccaactac taaactgggc cctgaagaag
      241 ggcccatata gggccctgaa gaagggcccc tcgactaagt ccaactacta aactgggccc
      301 tgaagaaggg cccatatagg gccctgaaga agggcccctc gactaagtcc aactactaaa
      361 ctgggccctg aagaagggcc catatagatc aagcttggta ccgagctcgg gccctgaaga
      421 agggcccctc gactaagtcc aactactaaa ctgggccctg aagaagggcc catatagggc
      481 cctgaagaag ggcccctcga ctaagtccaa ctactaaact gggccctgaa gaagggccca
      541 tatagggccc tgaagaaggg cccctcgact aagtccaact actaaactgg gccctgaaga
      601 agggcccata tatcgagtaa ggatctccag gcatcaaata aaacgaaagg ctcagtcgaa
      661 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac
      721 tggctcacct tcgggtgggc ctttctgcgt ttatacctag ggatatattc cgcttcctcg
      781 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
      841 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
      901 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
      961 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct
     1021 ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc
     1081 cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg
     1141 gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt
     1201 cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga
     1261 tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt
     1321 tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac
     1381 cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag
     1441 accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt
     1501 gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata agttgttact
     1561 agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
     1621 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat
     1681 caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     1741 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     1801 tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat
     1861 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca
     1921 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     1981 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     2041 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     2101 gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa
     2161 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     2221 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     2281 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     2341 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     2401 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc
     2461 gccagatatc gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata
     2521 tgatcaattc aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga
     2581 tagcttgtcg taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag
     2641 cgacttgatg ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga
     2701 gtgcatataa tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt
     2761 cgagagtttc atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc
     2821 gatgacttag taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc
     2881 tttccccttc taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg
     2941 cgtcgagcaa agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta
     3001 gcttctgggc gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta
     3061 atgcgctgtt aatcacttta cttttatcta atctagacat cattaattcc taatttttgt
     3121 tgacactcta tcgttgatag agttatttta ccactcccta tcagtgatag agaaaagaat
     3181 tcggtctca
//
