LOCUS       sequence-361842-        3425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of lambda-CI fused via three alanines to the
            MS2 coat protein. Useful as RNA-DNA adapter in three-hybrid system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3425)
  AUTHORS   Wang CD, Mansky R, LeBlanc H, Gravel CM, Berry KE
  TITLE     Optimization of a bacterial three-hybrid assay through in vivo 
            titration of an RNA-DNA adapter protein.
  JOURNAL   RNA. 2021 Apr;27(4):513-526. doi: 10.1261/rna.077404.120. Epub 2021 
            Jan 26.
  PUBMED    33500316
REFERENCE   2  (bases 1 to 3425)
  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: 
            "10.1261/rna.077404.120"; journalName: "RNA"; date: "2021-04"; 
            volume: "27"; issue: "4"; pages: "513-526"
COMMENT     Sequence Label: p35u4
FEATURES             Location/Qualifiers
     source          1..3425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(714..731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(848..1393)
                     /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(928..947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1507..2217
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             complement(join(2985..3425,1..219))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgag
     1441 gcctcgagac agatgctagc tcagtcctag gtatagtgct agcgcatgcc acacaggaaa
     1501 cagcgtatga gcacaaaaaa gaaaccatta acacaagagc agcttgagga cgcacgtcgc
     1561 cttaaagcaa tttatgaaaa aaagaaaaat gaacttggct tatcccagga atctgtcgca
     1621 gacaagatgg ggatggggca gtcaggcgtt ggtgctttat ttaatggcat caatgcatta
     1681 aatgcttata acgccgcatt gcttgcaaaa attctcaaag ttagcgttga agaatttagc
     1741 ccttcaatcg ccagagaaat ctacgagatg tatgaagcgg ttagtatgca gccgtcactt
     1801 agaagtgagt atgagtaccc tgttttttct catgttcagg cagggatgtt ctcacctgag
     1861 cttagaacct ttaccaaagg tgatgcggag agatgggtaa gcacaaccaa aaaagccagt
     1921 gattctgcat tctggcttga ggttgaaggt aattccatga ccgcaccaac aggctccaag
     1981 ccaagctttc ctgacggaat gttaattctc gttgaccctg agcaggctgt tgagccaggt
     2041 gatttctgca tagccagact tgggggtgat gagtttacct tcaagaaact gatcagggat
     2101 agcggtcagg tgtttttaca accactaaac ccacagtacc caatgatccc atgcaatgag
     2161 agttgttccg ttgtggggaa agttatcgct agtcagtggc ctgaagagac gtttggcgcg
     2221 gccgcagaat tcccggggat tcacccgggt atggcttcta actttactca gttcgttctc
     2281 gtcgacaatg gcggaactgg cgacgtgact gtcgccccaa gcaacttcgc taacgggatc
     2341 gctgaatgga tcagctctaa ctcgcgttca caggcttaca aagtaacctg tagcgttcgt
     2401 cagagctctg cgcagaatcg caaatacacc atcaaagtcg aggtgcctaa aggcgcctgg
     2461 cgttcgtact taaatatgga actaaccatt ccaattttcg ccacgaattc cgactgcgag
     2521 cttattgtta aggcaatgca aggtctccta aaagatggaa acccgattcc ctcagcaatc
     2581 gcagcaaact ccggcatcta ctaaggatcc ttaccttaca gatctgcatc gcaggatgct
     2641 gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctaaccg tttttatcag
     2701 gctctgggag gcagaataaa tgatcatatc gtcaattatt acctccacgg ggagagcctg
     2761 agcaaactgg cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa
     2821 accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac
     2881 cgggtcgaat ttgctttcga atttctgcca ttcatccgct tattatcact tattcaggcg
     2941 tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc
     3001 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     3061 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     3121 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     3181 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     3241 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     3301 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     3361 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     3421 atacg
//
