LOCUS       sequence-382579-        3398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of tdMCP from a lacZ-alpha inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3398)
  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 3398)
  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: placZ-alpha-tdMCP
FEATURES             Location/Qualifiers
     source          1..3398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    23..41
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    48..66
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             85..96
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(177..194)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(177..193)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(186..208)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(402..421)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             478..864
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2
                     /note="binds to a specific stem-loop structure in the viral
                     RNA (Peabody, 1993)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQNRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     misc_feature    1266..1286
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1287..1344
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1368..1385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1539..2127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1619..1638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2298..3089,3090..3158))
                     /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     2921..2940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3159..3263)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3331..3349
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3353..3396)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agtccctatc agtgatagag attgacatcc ctatcagtga
       61 tagagatact gagcactact agagaaagag gagaaatact agatgaccat gattacgcca
      121 agcttgcatg catgcaggtc gacgctagag gatccccggg taccgagctc aaattcactg
      181 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
      241 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
      301 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
      361 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
      421 gcagcggtct caaccggagg tggtggcagt accagcggtg gaggcggttc aaccatggca
      481 tccaacttca cgcaattcgt gttagtagac aatggcggca ctggggatgt gacagtggcg
      541 ccctcaaact ttgcaaatgg tgtagcagag tggatttcga gtaattctcg ctcgcaagct
      601 tataaagtga cttgttcggt acgtcagagc tcggcccaaa atcgtaaata tacgatcaag
      661 gtcgaagtac ccaaagtcgc gactcaaacc gtagggggcg ttgagttgcc tgtcgccgcc
      721 tggcgtagtt accttaatat ggagcttacg attcccattt ttgcgacaaa ttcggattgc
      781 gaattaattg tgaaagccat gcaagggctg ctgaaggacg gtaatcccat cccgagcgca
      841 attgccgcta actctggcat ttatgcgaat tttacccagt ttgttttggt cgataatggt
      901 ggtactggtg atgttactgt agcgccttca aactttgcga atggagtcgc agagtggatc
      961 agctcgaact cgcgctccca agcatataag gtgacgtgta gtgttcgtca aagctccgcg
     1021 cagaaccgta agtacactat taaagtcgag gtgcccaagg tcgccaccca gacggtcgga
     1081 ggggttgaat tacctgtagc agcctggcgt tcttatctta acatggagtt gactatccct
     1141 atcttcgcta cgaactctga ttgtgaatta atcgttaagg ccatgcaagg cctgttaaag
     1201 gacggaaatc ccatcccctc cgctattgca gccaattccg gcatctacta ataaattctg
     1261 agacctacta gtagcggccg ctgcagtccg gcaaaaaagg gcaaggtgtc accaccctgc
     1321 cctttttctt taaaaccgaa aagattactt cgcgttatgc aggcttcctc gctcactgac
     1381 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1441 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1501 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccacaggct ccgcccccct
     1561 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1621 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1681 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1741 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1801 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1861 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1921 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1981 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2041 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2101 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2161 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2221 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2281 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2341 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2401 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     2461 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2521 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     2581 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     2641 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2701 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2761 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     2821 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     2881 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     2941 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3001 ttaccgctgt tgagatccag ttcgatataa cccactcgtg cacccaactg atcttcagca
     3061 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3121 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3181 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3241 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     3301 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggca gaatttcaga
     3361 taaaaaaaat ccttagcttt cgctaaggat gatttctg
//
