LOCUS       sequence-490655-        3543 bp    DNA     circular SYN 24-OCT-2025
DEFINITION  template plasmid for site-saturation mutagenesis library cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3543)
  AUTHORS   Li S, Lee B, Lao Y, Lertwiriyapiti S, Huang X, Wang T
  TITLE     Directed evolution of a plant Rubisco chaperone with altered client 
            recognition.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 Sep 16;122(37):e2510701122. doi: 
            10.1073/pnas.2510701122. Epub 2025 Sep 11.
  PUBMED    40932770
REFERENCE   2  (bases 1 to 3543)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2025-09-16"; volume: "122(37):e2510701122. 
            doi"; pages: " 10.1073/pnas.2510701122. Epub 2025 Sep 11"
COMMENT     Sequence Label: pSYL017
FEATURES             Location/Qualifiers
     source          1..3543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     489..508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     742..759
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    774..914
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(840..859)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1000..1022
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1016..1207)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKPALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2651..3442)
                     /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"
     promoter        complement(3443..3534)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(3543,1..588)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
       61 gcggtggttt gtttgccgga tcaagagcta ccacctcttt ttccgaaggt aactggcttc
      121 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
      181 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      241 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      301 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      361 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      421 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      481 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      541 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      601 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
      661 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
      721 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
      781 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
      841 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
      901 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      961 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1021 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     1081 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     1141 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcgggt ttttcctgtt
     1201 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     1261 aacgagagag gatgctcacg atacgggtta ctgatgatga acaagaggac atccggtcaa
     1321 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttgctgagga gacttaggga
     1381 ccctggtctc tgggtatgca acagctctac caaccgttcc gaccgccgtc atctcccatt
     1441 ccaacccaat tccgttcact cgactccgcc ggtaaaatcg aagtcctcgc cggtcgaatg
     1501 gctctctggt tcgaatacgc acctcttatc tcttctcttt acaccgatgg tttcactcct
     1561 ccaaccatcg aagaactcac cggaatctca agcttcgaac agaaccgttt aatcgtcggt
     1621 gcgcaagttc atgactcaat tcttcaatct atccatgaac cggaacttat ttctgcgttc
     1681 gacaccggtg gtgcagagct tctatacgaa attcgtctcc ttagcaccac gcagcgtgtt
     1741 gcagccgcca cattcatcat tgatcgtaat attgattcga aaggagcaca ggatttggca
     1801 cgagccatca aggattatcc taaccgtcgt ggagacgttg gatggttgga ttttgattac
     1861 aacttaccag gagattgtct ttcattcttg tattacaggc agagcagaga gaataagaat
     1921 ccatcggatc agagaacctc gaagcttcta caagcattgg gggttgcaga atcagaaaag
     1981 gctaagaaca ggttgaacac agagctatac ggggacaagg aagcagagaa ggagaaggtg
     2041 aagaagaaga aggaagagga agtcaaagct attcggattc cagtggtgag gcttaagttt
     2101 ggagaagtag cagaagcgac ctcggtggta gttttaccgg tttgtaaagc agaggaaggg
     2161 gaaaagaaga ttcttgaagc tccaatggag atcatagccg gaggggattt taaggcggtt
     2221 gaggcggaga aaggatggaa gagatgggta gtgcttccgt cgtggaaccc agtggcagcc
     2281 attgggaaag gcggtgtggc ggtttctttc agggatgata gaaaagtgct gccttgggat
     2341 ggaaaggagg agcctttact ggtagtggcc gatagggtga ggaatgttgt ggaggctgat
     2401 gacgggtatt atctcgtggt ggctgagaac ggacttaagc tagagatagg atcagatttg
     2461 aaggcgagag aggtgaagga gagtttaggg atggttgttt tggtggtggg gccgccaaga
     2521 gaagatgatg atgattggca gacaagtcat cagaactggg actgaattaa gagaccatta
     2581 acggcactcc tcagcaaata taatgaccct cttgataacc caagagggca ttttttaatg
     2641 cccatggcgt ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt
     2701 cttccaactg atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc
     2761 tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag
     2821 cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa
     2881 gcactacatt tcgctcatct ccagcccagt cgggcggcga gttccatagc gttaaggttt
     2941 catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg
     3001 gacctaccaa ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt
     3061 cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt
     3121 gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga
     3181 cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt
     3241 tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa
     3301 tatcactgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca
     3361 acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt
     3421 cggcgatcac cgcttccctc atactcttcc tttttcaata ttattgaagc atttatcagg
     3481 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaatagg ccaaataggc
     3541 cgt
//
