LOCUS       sequence-83914-e        3853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of lambda-CI fused via three alanines to E. coli RNAP 
            beta subunit, aa 831-1057 (beta-flap). Can be cut with Not1 and 
            BamHI to replace beta-flap with a moiety of interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3853)
  AUTHORS   Dove SL, Joung JK, Hochschild A
  TITLE     Activation of prokaryotic transcription through arbitrary 
            protein-protein contacts.
  JOURNAL   Nature. 1997 Apr 10;386(6625):627-30.
  PUBMED    9121589
REFERENCE   2  (bases 1 to 3853)
  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"; journalName: "Nature."; 
            date: "1997-04-10"; volume: "386(6625)"; pages: "627-30"
COMMENT     Sequence Label: pAC-lambda-CI-beta-flap (831-1057)
FEATURES             Location/Qualifiers
     source          1..3853
                     /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..1392)
                     /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"
     promoter        join(1555..1560,1561..1578,1579..1585)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1593..1609
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1598..1620
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1630..2340
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             complement(join(3413..3853,1..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 cgggtgaatg ctgccaactt actgattagt 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 gtttcccctg gcggctccct
      961 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg
     1021 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact
     1081 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg
     1141 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta
     1201 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt
     1261 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc
     1321 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca
     1381 aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat ttcagtgcaa
     1441 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca
     1501 tgtttgacag cttatcatcg ataagctaat tctcactcat taggcacccc aggctttaca
     1561 ctttatgctt ccggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg
     1621 aaacagcgta tgagcacaaa aaagaaacca ttaacacaag agcagcttga ggacgcacgt
     1681 cgccttaaag caatttatga aaaaaagaaa aatgaacttg gcttatccca ggaatctgtc
     1741 gcagacaaga tggggatggg gcagtcaggc gttggtgctt tatttaatgg catcaatgca
     1801 ttaaatgctt ataacgccgc attgcttgca aaaattctca aagttagcgt tgaagaattt
     1861 agcccttcaa tcgccagaga aatctacgag atgtatgaag cggttagtat gcagccgtca
     1921 cttagaagtg agtatgagta ccctgttttt tctcatgttc aggcagggat gttctcacct
     1981 gagcttagaa cctttaccaa aggtgatgcg gagagatggg taagcacaac caaaaaagcc
     2041 agtgattctg cattctggct tgaggttgaa ggtaattcca tgaccgcacc aacaggctcc
     2101 aagccaagct ttcctgacgg aatgttaatt ctcgttgacc ctgagcaggc tgttgagcca
     2161 ggtgatttct gcatagccag acttgggggt gatgagttta ccttcaagaa actgatcagg
     2221 gatagcggtc aggtgttttt acaaccacta aacccacagt acccaatgat cccatgcaat
     2281 gagagttgtt ccgttgtggg gaaagttatc gctagtcagt ggcctgaaga gacgtttggc
     2341 gcggccgcaa tccacattca ggaactggcg tgtgtgtccc gtgacaccaa gctgggtccg
     2401 gaagagatca ccgctgacat cccgaacgtg ggtgaagctg cgctctccaa actggatgaa
     2461 tccggtatcg tttacattgg tgcggaagtg accggtggcg acattctggt tggtaaggta
     2521 acgccgaaag gtgaaactca gctgacccca gaagaaaaac tgctgcgtgc gatcttcggt
     2581 gagaaagcct ctgacgttaa agactcttct ctgcgcgtac caaacggtgt atccggtacg
     2641 gttatcgacg ttcaggtctt tactcgcgat ggcgtagaaa aagacaaacg tgcgctggaa
     2701 atcgaagaaa tgcagctcaa acaggcgaag aaagacctgt ctgaagaact gcagatcctc
     2761 gaagcgggtc tgttcagccg tatccgtgct gtgctggtag ccggtggcgt tgaagctgag
     2821 aagctcgaca aactgccgcg cgatcgctgg ctggagctgg gcctgacaga cgaagagaaa
     2881 caaaatcagc tggaacagct ggctgagcag tatgacgaac tgaaacacga gttcgagaag
     2941 aaactcgaag cgaaacgccg caaaatcacc cagggcgacg atctggcacc gggcgtgctg
     3001 aagattgtta aggtatatct ggcggttaaa taaggatcct taccttacag atctgcatcg
     3061 caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctaaccgt
     3121 ttttatcagg ctctgggagg cagaataaat gatcatatcg tcaattatta cctccacggg
     3181 gagagcctga gcaaactggc ctcaggcatt tgagaagcac acggtcacac tgcttccggt
     3241 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
     3301 accgacgacc gggtcgaatt tgctttcgaa tttctgccat tcatccgctt attatcactt
     3361 attcaggcgt agcaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc
     3421 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc
     3481 catcacagac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
     3541 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
     3601 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
     3661 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
     3721 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
     3781 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
     3841 tcattgccat acg
//
