LOCUS       sequence-382576-        3955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription of 12xMS2 repeats from a tetracycline-inducible 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3955)
  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 3955)
  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: pMS2
FEATURES             Location/Qualifiers
     source          1..3955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        73..91
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        112..130
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        182..200
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        221..239
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        291..309
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        330..348
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        400..418
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        439..457
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        509..527
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        548..566
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        618..636
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        657..675
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    781..1369
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(834..854)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1252..1263)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1372..1388)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1373..1389)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      1408..1479
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1495..1522
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1550..1567)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1684..2229)
                     /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(1764..1783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2343..2437)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2458..3117)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3032..3051
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3118..3220)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(3250..3876)
                     /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     3781..3800
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        3892..3947
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    3928..3946
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 agacggagct cgtcgacgcg gccgcaagct tggtaccgag ctcggatcct aaggtaccta
       61 attgcctaga aaacatgagg atcacccatg tctgcaggtc gactctagaa aacatgagga
      121 tcacccatgt ctgcagtatt cccgggttca ttagatccta aggtacctaa ttgcctagaa
      181 aacatgagga tcacccatgt ctgcaggtcg actctagaaa acatgaggat cacccatgtc
      241 tgcagtattc ccgggttcat tagatcctaa ggtacctaat tgcctagaaa acatgaggat
      301 cacccatgtc tgcaggtcga ctctagaaaa catgaggatc acccatgtct gcagtattcc
      361 cgggttcatt agatcctaag gtacctaatt gcctagaaaa catgaggatc acccatgtct
      421 gcaggtcgac tctagaaaac atgaggatca cccatgtctg cagtattccc gggttcatta
      481 gatcctaagg tacctaattg cctagaaaac atgaggatca cccatgtctg caggtcgact
      541 ctagaaaaca tgaggatcac ccatgtctgc agtattcccg ggttcattag atcctaaggt
      601 acctaattgc ctagaaaaca tgaggatcac ccatgtctgc aggtcgactc cagaaaacat
      661 gaggatcacc catgtctgca gtattcccgg gttcattaga tctgcgcgcg atcgatatca
      721 gcgctttaaa tttgcgcatg ctagtgtaag gcatgcaagc ttgatatcaa gcttatcgat
      781 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
      841 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
      901 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
      961 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     1021 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     1081 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     1141 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     1201 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     1261 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     1321 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
     1381 cgacctcgag taaggatctc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     1441 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca
     1501 ccttcgggtg ggcctttctg cgtttatacc tagggatata ttccgcttcc tcgctcactg
     1561 actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag
     1621 atttcctgga agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc
     1681 gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg
     1741 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt
     1801 gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt
     1861 gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta
     1921 tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1981 ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag
     2041 gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac
     2101 tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa
     2161 aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa
     2221 cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt
     2281 atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgtt actagtgctt
     2341 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     2401 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta
     2461 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     2521 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     2581 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     2641 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     2701 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     2761 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     2821 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     2881 gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa gaatgtgaat
     2941 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     3001 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     3061 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     3121 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     3181 ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccagat
     3241 atcgacgtct taagacccac tttcacattt aagttgtttt tctaatccgc atatgatcaa
     3301 ttcaaggccg aataagaagg ctggctctgc accttggtga tcaaataatt cgatagcttg
     3361 tcgtaataat ggcggcatac tatcagtagt aggtgtttcc ctttcttctt tagcgacttg
     3421 atgctcttga tcttccaata cgcaacctaa agtaaaatgc cccacagcgc tgagtgcata
     3481 taatgcattc tctagtgaaa aaccttgttg gcataaaaag gctaattgat tttcgagagt
     3541 ttcatactgt ttttctgtag gccgtgtacc taaatgtact tttgctccat cgcgatgact
     3601 tagtaaagca catctaaaac ttttagcgtt attacgtaaa aaatcttgcc agctttcccc
     3661 ttctaaaggg caaaagtgag tatggtgcct atctaacatc tcaatggcta aggcgtcgag
     3721 caaagcccgc ttatttttta catgccaata caatgtaggc tgctctacac ctagcttctg
     3781 ggcgagttta cgggttgtta aaccttcgat tccgacctca ttaagcagct ctaatgcgct
     3841 gttaatcact ttacttttat ctaatctaga catcattaat tcctaatttt tgttgacact
     3901 ctatcgttga tagagttatt ttaccactcc ctatcagtga tagagaaaag aattc
//
