LOCUS       sequence-83920-e        5782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of alpha NTD fused via three alanines to E. coli RNAP 
            subunit beta, 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 5782)
  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 5782)
  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: pBR-alpha-beta flap (831-1057)
FEATURES             Location/Qualifiers
     source          1..5782
                     /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(1859..1878)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             3336..3527
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3521..3543)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3629..3769
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3684..3703
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3784..3801)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3955..4543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4035..4054)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4714..5505,5506..5574))
                     /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     5337..5356
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5575..5679)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5747..5765
                     /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 aatccacatt caggaactgg cgtgtgtgtc
     1141 ccgtgacacc aagctgggtc cggaagagat caccgctgac atcccgaacg tgggtgaagc
     1201 tgcgctctcc aaactggatg aatccggtat cgtttacatt ggtgcggaag tgaccggtgg
     1261 cgacattctg gttggtaagg taacgccgaa aggtgaaact cagctgaccc cagaagaaaa
     1321 actgctgcgt gcgatcttcg gtgagaaagc ctctgacgtt aaagactctt ctctgcgcgt
     1381 accaaacggt gtatccggta cggttatcga cgttcaggtc tttactcgcg atggcgtaga
     1441 aaaagacaaa cgtgcgctgg aaatcgaaga aatgcagctc aaacaggcga agaaagacct
     1501 gtctgaagaa ctgcagatcc tcgaagcggg tctgttcagc cgtatccgtg ctgtgctggt
     1561 agccggtggc gttgaagctg agaagctcga caaactgccg cgcgatcgct ggctggagct
     1621 gggcctgaca gacgaagaga aacaaaatca gctggaacag ctggctgagc agtatgacga
     1681 actgaaacac gagttcgaga agaaactcga agcgaaacgc cgcaaaatca cccagggcga
     1741 cgatctggca ccgggcgtgc tgaagattgt taaggtatat ctggcggtta aataaggatc
     1801 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
     1861 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
     1921 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
     1981 ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc
     2041 ttcctaatgc aggagtcgca taagggagag cgtcgaccga tgcccttgag agccttcaac
     2101 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     2161 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag
     2221 gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg
     2281 cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag
     2341 gccattatcg ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg
     2401 cgaggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat cgggatgccc
     2461 gcgttgcagg ccatgctgtc caggcaggta gatgacgacc atcagggaca gcttcaagga
     2521 tcgctcgcgg ctcttaccag cctaacttcg atcactggac cgctgatcgt cacggcgatt
     2581 tatgccgcct cggcgagcac atggaacggg ttggcatgga ttgtaggcgc cgccctatac
     2641 cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc gacctgaatg
     2701 gaagccggcg gcacctcgct aacggattca ccactccaag aattggagcc aatcaattct
     2761 tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa catatccatc gcgtccgcca
     2821 tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca cgggtgcgca
     2881 tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc
     2941 agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga
     3001 cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga
     3061 agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct
     3121 gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct
     3181 ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat
     3241 gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc
     3301 ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc
     3361 ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc
     3421 tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt
     3481 accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     3541 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     3601 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     3661 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     3721 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
     3781 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3841 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3901 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3961 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4021 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4081 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4141 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4201 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4261 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4321 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4381 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4441 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4501 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4561 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4621 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4681 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4741 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4801 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4861 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4921 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4981 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt
     5041 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5101 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5161 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5221 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5281 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa
     5341 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5401 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5461 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5521 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5581 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5641 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5701 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5761 gaggcccttt ggataaccag aa
//
