LOCUS       sequence-340792-        3307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modified pET17b (Novagen) vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3307)
  AUTHORS   Daniels C, Morona R
  TITLE     Analysis of Shigella flexneri wzz (Rol) function by mutagenesis and 
            cross-linking: wzz is able to oligomerize.
  JOURNAL   Mol Microbiol. 1999 Oct;34(1):181-94. doi: 
            10.1046/j.1365-2958.1999.01591.x.
  PUBMED    10540296
REFERENCE   2  (bases 1 to 3307)
  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: 
            "10.1046/j.1365-2958.1999.01591"; journalName: "Mol Microbiol"; 
            date: "1999-10"; volume: "34"; issue: "1"; pages: "181-94"
COMMENT     Sequence Label: pRMCD77
FEATURES             Location/Qualifiers
     source          1..3307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(48..95)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     91..109
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(257..289)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     RBS             297..319
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(350..369)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(351..369)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             882..1073
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1067..1089)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1175..1314
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1230..1249
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1329..1346)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1500..2088)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1580..1599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2259..3050,3051..3119))
                     /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     2882..2901
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3120..3224)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3292..3307,1..3)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggccctttcg tcttcaagaa ttatccggat atagttcctc ctttcagcaa aaaacccctc
       61 aagacccgtt tagaggcccc aaggggttat gctagttatt gctcagcggt ggcagcagcc
      121 aactcagctt cctttcgggc tttgttagca gccggatctg ctcgagcggc cgccagtgtg
      181 atggatatct gcagaattcc agcacactgg cggccgttac tagtggatcc gagctcggta
      241 ccaagcttga atcccgaccc atttgctgtc caccagtcat gctagccata tgtatatctc
      301 cttcttaaag ttaaacaaaa ttatttctag agggaaaccg ttgtggtctc cctatagtga
      361 gtcgtattaa tttcgcggga tcgagatcga tctcgggcag cgttgggtcc tggccacggg
      421 tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg
      481 gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt
      541 ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa
      601 cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc
      661 taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt
      721 ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc
      781 gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca
      841 ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa
      901 ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca
      961 acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg
     1021 agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     1081 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     1141 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     1201 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     1261 ccattgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct
     1321 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1381 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1441 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1501 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1561 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1621 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1681 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1741 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1801 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1861 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1921 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1981 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2041 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2101 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2161 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2221 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2281 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2341 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2401 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2461 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2521 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc
     2581 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     2641 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     2701 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     2761 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     2821 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     2881 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     2941 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3001 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3061 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3121 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3181 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3241 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     3301 atcacga
//
