LOCUS       sequence-382526-        3472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of 47 rCAG repeats fused to a translational unit 
            over-expressing superfolder GFP using a strong RBS. The RBS contains
            mutation (m1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3472)
  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 3472)
  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-RBS""m1""-sfGFP
FEATURES             Location/Qualifiers
     source          1..3472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      918..989
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1005..1032
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1060..1077)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1194..1739)
                     /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(1274..1293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1853..1947)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1968..2627)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2542..2561
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2628..2730)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(2760..3386)
                     /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     3291..3310
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        3402..3457
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    3438..3456
                     /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 aaagaaatga
      181 gaatactagt actagatgag caaaggcgaa gaactgttta ccggcgtggt gccgattctg
      241 gttgaactgg atggcgatgt taatggccat aagtttagcg tgcgcggcga aggcgaaggc
      301 gatgcgacca acggcaaact gaccctgaag tttatttgca ccaccggcaa actgccggtg
      361 ccgtggccga ccctggtgac caccctgacc tatggcgtgc agtgctttag ccgctatccg
      421 gatcacatga aacgccatga tttctttaag tccgcgatgc cggaaggcta tgttcaggaa
      481 cgcaccatta gctttaaaga tgatggcacc tataagaccc gcgcggaagt gaaatttgaa
      541 ggcgataccc tggtgaaccg cattgaactg aaaggcattg acttcaaaga agatggcaac
      601 attctgggcc ataaactgga gtacaatttc aacagccata acgtgtatat taccgcggac
      661 aaacagaaga acggcatcaa agcgaatttc aaaatccgcc ataatgtgga agatggcagc
      721 gtgcagctgg cggatcacta tcagcagaat accccgatcg gcgatggccc ggtgctgctg
      781 ccggacaatc actacctgag cacccagtcc gtgctgagca aagatccgaa tgagaaacgc
      841 gatcacatgg ttctgctgga gtttgtgacc gcggcgggta tcactcatgg ctaactcgag
      901 taaggatctc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
      961 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     1021 ggcctttctg cgtttatacc tagggatata ttccgcttcc tcgctcactg actcgctacg
     1081 ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga
     1141 agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat
     1201 aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac
     1261 ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct
     1321 gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc
     1381 acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc
     1441 ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga
     1501 aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct
     1561 tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc
     1621 caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct
     1681 gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag
     1741 aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt atctcttcaa
     1801 atgtagcacc tgaagtcagc cccatacgat ataagttgtt actagtgctt ggattctcac
     1861 caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg
     1921 tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc
     1981 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     2041 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     2101 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     2161 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     2221 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     2281 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     2341 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     2401 gccatacgaa attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     2461 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     2521 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     2581 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     2641 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     2701 tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccagat atcgacgtct
     2761 taagacccac tttcacattt aagttgtttt tctaatccgc atatgatcaa ttcaaggccg
     2821 aataagaagg ctggctctgc accttggtga tcaaataatt cgatagcttg tcgtaataat
     2881 ggcggcatac tatcagtagt aggtgtttcc ctttcttctt tagcgacttg atgctcttga
     2941 tcttccaata cgcaacctaa agtaaaatgc cccacagcgc tgagtgcata taatgcattc
     3001 tctagtgaaa aaccttgttg gcataaaaag gctaattgat tttcgagagt ttcatactgt
     3061 ttttctgtag gccgtgtacc taaatgtact tttgctccat cgcgatgact tagtaaagca
     3121 catctaaaac ttttagcgtt attacgtaaa aaatcttgcc agctttcccc ttctaaaggg
     3181 caaaagtgag tatggtgcct atctaacatc tcaatggcta aggcgtcgag caaagcccgc
     3241 ttatttttta catgccaata caatgtaggc tgctctacac ctagcttctg ggcgagttta
     3301 cgggttgtta aaccttcgat tccgacctca ttaagcagct ctaatgcgct gttaatcact
     3361 ttacttttat ctaatctaga catcattaat tcctaatttt tgttgacact ctatcgttga
     3421 tagagttatt ttaccactcc ctatcagtga tagagaaaag aattcggtct ca
//
