LOCUS       sequence-361824-        3163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of the lambda-CI protein; used as 
            three-hybrid control corresponding to pCW17 (addgene #174664).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3163)
  AUTHORS   Stockert OM, Gravel CM, Berry KE
  TITLE     A bacterial three-hybrid assay for forward and reverse genetic 
            analysis of RNA-protein interactions.
  JOURNAL   Nat Protoc. 2022 Apr;17(4):941-961. doi: 10.1038/s41596-021-00657-4.
            Epub 2022 Feb 23.
  PUBMED    35197605
REFERENCE   2  (bases 1 to 3163)
  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: 
            "10.1038/s41596-021-00657-4"; journalName: "Nat Protoc"; date: 
            "2022-04"; volume: "17"; issue: "4"; pages: "941-961"
COMMENT     Sequence Label: pRM16
FEATURES             Location/Qualifiers
     source          1..3163
                     /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             1620..2330
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             complement(join(2723..3163,1..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 tttcagtgca
     1441 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     1501 atgtttgaca gcttatcatc gataagctaa ttctcactca ttaggcaccc caggcctcga
     1561 gacgatagct agctcagtcc taggtatagt gctagcgcat gccacacagg aaacagcgta
     1621 tgagcacaaa aaagaaacca ttaacacaag agcagcttga ggacgcacgt cgccttaaag
     1681 caatttatga aaaaaagaaa aatgaacttg gcttatccca ggaatctgtc gcagacaaga
     1741 tggggatggg gcagtcaggc gttggtgctt tatttaatgg catcaatgca ttaaatgctt
     1801 ataacgccgc attgcttgca aaaattctca aagttagcgt tgaagaattt agcccttcaa
     1861 tcgccagaga aatctacgag atgtatgaag cggttagtat gcagccgtca cttagaagtg
     1921 agtatgagta ccctgttttt tctcatgttc aggcagggat gttctcacct gagcttagaa
     1981 cctttaccaa aggtgatgcg gagagatggg taagcacaac caaaaaagcc agtgattctg
     2041 cattctggct tgaggttgaa ggtaattcca tgaccgcacc aacaggctcc aagccaagct
     2101 ttcctgacgg aatgttaatt ctcgttgacc ctgagcaggc tgttgagcca ggtgatttct
     2161 gcatagccag acttgggggt gatgagttta ccttcaagaa actgatcagg gatagcggtc
     2221 aggtgttttt acaaccacta aacccacagt acccaatgat cccatgcaat gagagttgtt
     2281 ccgttgtggg gaaagttatc gctagtcagt ggcctgaaga gacgtttggc gcggccgcat
     2341 aaggatcctt accttacaga tctgcatcgc aggatgctgc tggctaccct gtggaacacc
     2401 tacatctgta ttaacgaagc gctaaccgtt tttatcaggc tctgggaggc agaataaatg
     2461 atcatatcgt caattattac ctccacgggg agagcctgag caaactggcc tcaggcattt
     2521 gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca
     2581 taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat
     2641 ttctgccatt catccgctta ttatcactta ttcaggcgta gcaaccaggc gtttaagggc
     2701 accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta
     2761 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     2821 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     2881 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     2941 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     3001 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     3061 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     3121 cactatccca tatcaccagc tcaccgtctt tcattgccat acg
//
