LOCUS       sequence-493006-        5572 bp    DNA     circular SYN 18-NOV-2025
DEFINITION  Express anti-sigma protein, AsiA, in E. coli. His-Tag for Western 
            Blots or Purification..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5572)
  AUTHORS   Weng X, Bohrer CH, Bettridge K, Lagda AC, Cagliero C, Jin DJ, Xiao J
  TITLE     Spatial organization of RNA polymerase and its relationship with 
            transcription in Escherichia coli.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Oct 1;116(40):20115-20123. doi: 
            10.1073/pnas.1903968116. Epub 2019 Sep 16.
  PUBMED    31527272
REFERENCE   2  (bases 1 to 5572)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1073/pnas.1903968116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-10-1- 1"; volume: "116"; issue: "40"; pages: 
            "20115-20123"
COMMENT     Sequence Label: pDR-pBAD-ATG-6xHIS-AsiA
FEATURES             Location/Qualifiers
     source          1..5572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      309..531
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             579..1529
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             2781..3572
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             complement(4123..4140)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4155..4177
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(4204..4488)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(4262..4281)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             4515..5393
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 acgccctgag aagcccgtga cgggcttttc ttgtattatg ggtagtttcc ttgcatgaat
       61 ccataaaagg cgcctgtagt gccatttacc cccattcact gccagagccg tgagcgcagc
      121 gaactgaatg tcacgaaaaa gacagcgact caggtgcctg atggtcggag acaaaaggaa
      181 tattcagcga tttgcccgag cttgcgaggg tgctacttaa gcctttaggg ttttaaggtc
      241 tgttttgtag aggagcaaac agcgtttgcg acatcctttt gtaatactgc ggaactgact
      301 aaagtagtga gttatacaca gggctgggat ctattctttt tatctttttt tattctttct
      361 ttattctata aattataacc acttgaatat aaacaaaaaa aacacacaaa ggtctagcgg
      421 aatttacaga gggtctagca gaatttacaa gttttccagc aaaggtctag cagaatttac
      481 agatacccac aactcaaagg aaaaggacta gtaattatca ttgactagcc catctcaatt
      541 ggtatagtga ttaaaatcac ctagaccaat tgagatgtat gtctgaatta gttgttttca
      601 aagcaaatga actagcgatt agtcgctatg acttaacgga gcatgaaacc aagctaattt
      661 tatgctgtgt ggcactactc aaccccacga ttgaaaaccc tacaaggaaa gaacggacgg
      721 tatcgttcac ttataaccaa tacgctcaga tgatgaacat cagtagggaa aatgcttatg
      781 gtgtattagc taaagcaacc agagagctga tgacgagaac tgtggaaatc aggaatcctt
      841 tggttaaagg ctttgagatt ttccagtgga caaactatgc caagttctca agcgaaaaat
      901 tagaattagt ttttagtgaa gagatattgc cttatctttt ccagttaaaa aaattcataa
      961 aatataatct ggaacatgtt aagtcttttg aaaacaaata ctctatgagg atttatgagt
     1021 ggttattaaa agaactaaca caaaagaaaa ctcacaaggc aaatatagag attagccttg
     1081 atgaatttaa gttcatgtta atgcttgaaa ataactacca tgagtttaaa aggcttaacc
     1141 aatgggtttt gaaaccaata agtaaagatt taaacactta cagcaatatg aaattggtgg
     1201 ttgataagcg aggccgcccg actgatacgt tgattttcca agttgaacta gatagacaaa
     1261 tggatctcgt aaccgaactt gagaacaacc agataaaaat gaatggtgac aaaataccaa
     1321 caaccattac atcagattcc tacctacata acggactaag aaaaacacta cacgatgctt
     1381 taactgcaaa aattcagctc accagttttg aggcaaaatt tttgagtgac atgcaaagta
     1441 agtatgatct caatggttcg ttctcatggc tcacgcaaaa acaacgaacc acactagaga
     1501 acatactggc taaatacgga aggatctgag gttcttatgg ctcttgtatc tatcagtgaa
     1561 gcatcaagac taacaaacaa aagtagaaca actgttcacc gttacatatc aaagggaaaa
     1621 ctgtccatat gcacagatga aaacggtgta aaaaagatag atacatcaga gcttttacga
     1681 gtttttggtg cattcaaagc tgttcaccat gaacagatcg acaatgtaac agatgaacag
     1741 catgtaacac ctaatagaac aggtgaaacc agtaaaacaa agcaactaga acatgaaatt
     1801 gaacacctga gacaacttgt tacagctcaa cagtcacaca tagacagcct gaaacaggcg
     1861 atgctgctta tcgaatcaaa gctgccgaca acacgggagc cagtgacgcc tcccgtgggg
     1921 aaaaaatcat ggcaattctg gaagaaatag cgctttcagc cggcaaaccg gctgaagccg
     1981 gatctgcgat tctgataaca aactagcaac accagaacag cccgtttgcg ggcagcaaaa
     2041 cccgtgggaa ttaattcccc tgctcgcgca ggctgggtgc caagctctcg ggtaacatca
     2101 aggcccgatc cttggagccc ttgccctccc gcacgatgat cgtgccgtga tcgaaatcca
     2161 gatccttgac ccgcagttgc aaaccctcac tgatccgcat gcccgttcca tacagaagct
     2221 gggcgaacaa acgatgctcg ccttccagaa aaccgaggat gcgaaccact tcatccgggg
     2281 tcagcaccac cggcaagcgc cgcgacggcc gaggtcttcc gatctcctga agccagggca
     2341 gatccgtgca cagcaccttg ccgtagaaga acagcaaggc cgccaatgcc tgacgatgcg
     2401 tggagaccga aaccttgcgc tcgttcgcca gccaggacag aaatgcctcg acttcgctgc
     2461 tgcccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga
     2521 cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt
     2581 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     2641 atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
     2701 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc
     2761 ctaaaacaaa gttaaacatc atgagggaag cggtgatcgc cgaagtatcg actcaactat
     2821 cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt
     2881 acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg
     2941 tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt
     3001 cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg
     3061 acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc
     3121 gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct
     3181 tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct
     3241 ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga
     3301 actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt
     3361 ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg
     3421 agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac
     3481 aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga
     3541 aaggcgagat caccaaggta gtcggcaaat aatgtctaac aattcgttca agccgacgcc
     3601 gcttcgcggc gcggcttaac tcaagcgtta gatgcactaa gcacataatt gctcacagcc
     3661 aaactatcag gtcaagtctg cttttattat ttttaagcgt gcataataag ccctacacaa
     3721 attgggagat atatcatgaa aggctggctt tttcttgtta tcgcaatagt tggcgaagta
     3781 atcgcaacat ccgcattaaa atctagcgag ggctttacta agcttgcatg cctgcaggtc
     3841 gacctactgc agttatttgt tggtatacat ttccagatag cgatacacac cttcaaagcc
     3901 ttcgttaaat tcatcgatca gggttttttt gtcctcttgg gtcagttcgc tcacaatttt
     3961 gcgaaagcta ttctgattca gtttacgacc ttcatgggtc acgccaattt cattcagaaa
     4021 ggcaataaag tttgcacggt tttccacgat atcttcacgg ctaaatttga tcagaatgct
     4081 tgcaacggtg ataatttcac gcacggtatc aatgtttttg ttatgatgat gatgatgatg
     4141 catgctagcg tatatctcct tcttaaagtt aaacaaaatt atttctagcc caaaaaaacg
     4201 ggtatggaga aacagtagag agttgcgata aaaagcgtca ggtaggatcc gctaatctta
     4261 tggataaaaa tgctatggca tagcaaagtg tgacgccgtg caaataatca atgtggactt
     4321 ttttgccgtg attatagaca cttttgttac gcgtttttgt catggctttg gtcccgcttt
     4381 gttacagaat gcttttaata agcggggtta ccggtttggt tagcgagaag agccagtaaa
     4441 agacgcagtg acggcaatgt ctgatgcaat atggacaatt ggtttcttct ctgaatggcg
     4501 ggagtatgaa aagtatggct gaagcgcaaa atgatcccct gctgccggga tactcgttta
     4561 atgcccatct ggtggcgggt ttaacgccga ttgaggccaa cggttatctc gattttttta
     4621 tcgaccgacc gctgggaatg aaaggttata ttctcaatct caccattcgc ggtcaggggg
     4681 tggtgaaaaa tcagggacga gaatttgttt gccgaccggg tgatattttg ctgttcccgc
     4741 caggagagat tcatcactac ggtcgtcatc cggaggctcg cgaatggtat caccagtggg
     4801 tttactttcg tccgcgcgcc tactggcatg aatggcttaa ctggccgtca atatttgcca
     4861 atacggggtt ctttcgcccg gatgaagcgc accagccgca tttcagcgac ctgtttgggc
     4921 aaatcattaa cgccgggcaa ggggaagggc gctattcgga gctgctggcg ataaatctgc
     4981 ttgagcaatt gttactgcgg cgcatggaag cgattaacga gtcgctccat ccaccgatgg
     5041 ataatcgggt acgcgaggct tgtcagtaca tcagcgatca cctggcagac agcaattttg
     5101 atatcgccag cgtcgcacag catgtttgct tgtcgccgtc gcgtctgtca catcttttcc
     5161 gccagcagtt agggattagc gtcttaagct ggcgcgagga ccaacgtatc agccaggcga
     5221 agctgctttt gagcaccacc cggatgccta tcgccaccgt cggtcgcaat gttggttttg
     5281 acgatcaact ctatttctcg cgggtattta aaaaatgcac cggggccagc ccgagcgagt
     5341 tccgtgccgg ttgtgaagaa aaagtgaatg atgtagccgt caagttgtca taagaattcc
     5401 cgacagtaag acgggtaagc ctgttgatga taccgctgcc ttactgggtg cattagccag
     5461 tctgaatgac ctgtcacggg ataatccgaa gtggtcagac tggaaaatca gagggcagga
     5521 actgctgaac agcaaaaagt cagatagcac cacatagcag acccgccata aa
//
