LOCUS       sequence-455420-        4103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription of 47xCAG and 12xMS2 repeats from a 
            tetracycline-inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4103)
  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 4103)
  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-MS2
FEATURES             Location/Qualifiers
     source          1..4103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        214..232
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        253..271
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        323..341
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        362..380
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        432..450
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        471..489
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        541..559
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        580..598
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        650..668
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        689..707
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        759..777
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        798..816
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    922..1510
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(975..995)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1393..1404)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1513..1529)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1514..1530)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      1549..1620
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1636..1663
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1691..1708)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1825..2370)
                     /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(1905..1924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2484..2578)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2599..3258)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3173..3192
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3259..3361)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(3391..4017)
                     /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     3922..3941
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        4033..4088
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    4069..4087
                     /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 ggccgcaagc ttggtaccga
      181 gctcggatcc taaggtacct aattgcctag aaaacatgag gatcacccat gtctgcaggt
      241 cgactctaga aaacatgagg atcacccatg tctgcagtat tcccgggttc attagatcct
      301 aaggtaccta attgcctaga aaacatgagg atcacccatg tctgcaggtc gactctagaa
      361 aacatgagga tcacccatgt ctgcagtatt cccgggttca ttagatccta aggtacctaa
      421 ttgcctagaa aacatgagga tcacccatgt ctgcaggtcg actccagaaa acatgaggat
      481 cacccatgtc tgcagtattc ccgggttcat tagatcctaa ggtacctaat tgcctagaaa
      541 acatgaggat cacccatgtc tgcaggtcga ctctagaaaa catgaggatc acccatgtct
      601 gcagtattcc cgggttcatt agatcctaag gtacctaatt gcctagaaaa catgaggatc
      661 acccatgtct gcaggtcgac tctagaaaac atgaggatca cccatgtctg cagtattccc
      721 gggttcatta gatcctaagg tacctaattg cctagaaaac atgaggatca cccatgtctg
      781 caggtcgact ccagaaaaca tgaggatcac ccatgtctgc agtattcccg ggttcattag
      841 atctgcgcgc gatcgatatc agcgctttaa atttgcgcat gctagtgtaa ggcatgcaag
      901 cttgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
      961 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     1021 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     1081 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     1141 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     1201 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     1261 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     1321 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     1381 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     1441 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     1501 gcctccccgc atcgataccg tcgacctcga gtaaggatct ccaggcatca aataaaacga
     1561 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1621 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatac ctagggatat
     1681 attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa
     1741 tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag
     1801 tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga
     1861 aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1921 tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat
     1981 tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag
     2041 ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat
     2101 ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag
     2161 ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact
     2221 gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg
     2281 gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa
     2341 gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat
     2401 ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga
     2461 tataagttgt tactagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc
     2521 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca
     2581 agcgagctcg atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc
     2641 attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag
     2701 cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa
     2761 gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc
     2821 tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta
     2881 acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact
     2941 ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact
     3001 atcccatatc accagctcac cgtctttcat tgccatacga aattccggat gagcattcat
     3061 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt
     3121 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga
     3181 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc
     3241 agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     3301 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc
     3361 aacgtctcat tttcgccaga tatcgacgtc ttaagaccca ctttcacatt taagttgttt
     3421 ttctaatccg catatgatca attcaaggcc gaataagaag gctggctctg caccttggtg
     3481 atcaaataat tcgatagctt gtcgtaataa tggcggcata ctatcagtag taggtgtttc
     3541 cctttcttct ttagcgactt gatgctcttg atcttccaat acgcaaccta aagtaaaatg
     3601 ccccacagcg ctgagtgcat ataatgcatt ctctagtgaa aaaccttgtt ggcataaaaa
     3661 ggctaattga ttttcgagag tttcatactg tttttctgta ggccgtgtac ctaaatgtac
     3721 ttttgctcca tcgcgatgac ttagtaaagc acatctaaaa cttttagcgt tattacgtaa
     3781 aaaatcttgc cagctttccc cttctaaagg gcaaaagtga gtatggtgcc tatctaacat
     3841 ctcaatggct aaggcgtcga gcaaagcccg cttatttttt acatgccaat acaatgtagg
     3901 ctgctctaca cctagcttct gggcgagttt acgggttgtt aaaccttcga ttccgacctc
     3961 attaagcagc tctaatgcgc tgttaatcac tttactttta tctaatctag acatcattaa
     4021 ttcctaattt ttgttgacac tctatcgttg atagagttat tttaccactc cctatcagtg
     4081 atagagaaaa gaattcggtc tca
//
