LOCUS       sequence-361823-        5404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plpp/PlacUV5- directed synthesis of the alpha NTD fused to E. coli 
            Hfq. Useful as three-hybrid positive control..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5404)
  AUTHORS   Berry KE, Hochschild A
  TITLE     A bacterial three-hybrid assay detects Escherichia coli Hfq-sRNA 
            interactions in vivo.
  JOURNAL   Nucleic Acids Res. 2018 Jan 25;46(2):e12. doi: 10.1093/nar/gkx1086.
  PUBMED    29140461
REFERENCE   2  (bases 1 to 5404)
  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: "Nucleic 
            Acids Res."; date: "2018-01-25"; volume: "46(2):e12. doi"; pages: " 
            10.1093/nar/gkx1086"
COMMENT     Sequence Label: pKB817 (pBra-Hfq)
FEATURES             Location/Qualifiers
     source          1..5404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(235..240,241..258,259..265)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    273..289
                     /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     278..300
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     297..313
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     297..313
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1481..1500)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             2958..3149
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3143..3165)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3251..3391
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3306..3325
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3406..3423)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3577..4165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3657..3676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4336..5127,5128..5196))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4959..4978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5197..5301)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5369..5387
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcaataaaaa atcaaatcgg atttcactat ataatctcac tttatctaag atgaatccga
       61 tggaagcatc ctgttttctc tcaatttttt tatctaaaac ccagcgttcg atgcttcttt
      121 gagcgaacga tcaaaaataa gtgccttccc atcaaaaaaa tattgacaac ataaaaaact
      181 ttgtgttata cttgtaacgc tacatggaga ttaactcaat ctagctagag aggctttaca
      241 ctttatgctt ccggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg
      301 aaacagctat gaccatgatt acggattcac tggaactcta gaccaaagag aggacacaat
      361 gcagggttct gtgacagagt ttctaaaacc gcgcctggtt gatatcgagc aagtgagttc
      421 gacgcacgcc aaggtgaccc ttgagccttt agagcgtggc tttggccata ctctgggtaa
      481 cgcactgcgc cgtattctgc tctcatcgat gccgggttgc gcggtgaccg aggttgagat
      541 tgatggtgta ctacatgagt acagcaccaa agaaggcgtt caggaagata tcctggaaat
      601 cctgctcaac ctgaaagggc tggcggtgag agttcagggc aaagatgaag ttattcttac
      661 cttgaataaa tctggcattg gccctgtgac tgcagccgat atcacccacg acggtgatgt
      721 cgaaatcgtc aagccgcagc acgtgatctg ccacctgacc gatgagaacg cgtctattag
      781 catgcgtatc aaagttcagc gcggtcgtgg ttatgtgccg gcttctaccc gaattcattc
      841 ggaagaagat gagcgcccaa tcggccgtct gctggtcgac gcatgctaca gccctgtgga
      901 gcgtattgcc tacaatgttg aagcagcgcg tgtagaacag cgtaccgacc tggacaagct
      961 ggtcatcgaa atggaaacca acggcacaat cgatcctgaa gaggcgattc gtcgtgcggc
     1021 aaccattctg gctgaacaac tggaagcttt cgttgactta cgtgatgtac gtcagcctga
     1081 agtgaaagaa gagaaaccag aggcggccgc agctaagggg caatctttac aagatccgtt
     1141 cctgaacgca ctgcgtcggg aacgtgttcc agtttctatt tatttggtga atggtattaa
     1201 gctgcaaggg caaatcgagt cttttgatca gttcgtgatc ctgttgaaaa acacggtcag
     1261 ccagatggtt tacaagcacg cgatttctac tgttgtcccg tctcgcccgg tttctcatca
     1321 cagtaacaac gccggtggcg gtaccagcag taactaccat catggtagca gcgcgcagaa
     1381 tacttccgcg caacaggaca gcgaagaaac cgaataagga tcctctacgc cggacgcatc
     1441 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     1501 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
     1561 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt
     1621 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
     1681 cataagggag agcgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     1741 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     1801 gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc
     1861 gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc
     1921 ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg
     1981 gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc
     2041 cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg
     2101 tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc
     2161 agcctaactt cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc
     2221 acatggaacg ggttggcatg gattgtaggc gccgccctat accttgtctg cctccccgcg
     2281 ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg
     2341 ctaacggatt caccactcca agaattggag ccaatcaatt cttgcggaga actgtgaatg
     2401 cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc catctccagc agccgcacgc
     2461 ggcgcatctc gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt
     2521 tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac
     2581 gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa
     2641 tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca
     2701 ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg
     2761 tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata
     2821 ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc
     2881 gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc
     2941 cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta
     3001 tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg
     3061 cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc
     3121 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     3181 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     3241 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
     3301 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     3361 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     3421 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3481 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3541 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3601 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3661 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3721 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3781 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3841 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3901 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3961 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4021 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4081 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4141 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4201 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4261 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4321 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4381 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4441 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4501 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4561 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4621 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     4681 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4741 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4801 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4861 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4921 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     4981 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5041 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5101 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5161 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5221 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5281 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5341 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttggataacc
     5401 agaa
//
