LOCUS       sequence-484662-        2780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains an AraC gene to assemble a selection phage missing gene III
            with SapI Golden Gate Assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2780)
  TITLE     Phage-Assisted Evolution of Allosteric Protein Switches
REFERENCE   2  (bases 1 to 2780)
  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"
COMMENT     Sequence Label: PACE_SP_SD4_AraC
FEATURES             Location/Qualifiers
     source          1..2780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1073..1888)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1073..1888)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1796..1815
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1880..1899
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2010..2598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2499..2518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2752..2769
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agtaaggagg aaaaatgtcc gcggaagcgc agaacgatcc gctgctgccg ggctatagct
       61 ttaacgcgca tctggtggcg ggcctgaccc cgattgaagc gaacggctat ctggattttt
      121 ttattgatcg cccgctgggc atgaaaggct atattctgaa cctgaccatt cgcggccagg
      181 gcgtggtgaa aaaccagggc cgcgaatttg tgtgccgccc gggcgatatt ctgctgtttc
      241 cgccgggcga aattcatcat tatggccgcc atccggaagc gcgcgaatgg tatcatcagt
      301 gggtgtattt tcgcccgcgc gcgtattggc atgaatggct gaactggccg agcatttttg
      361 cgaacaccgg cttttttcgc ccggatgaag cgcatcagcc gcattttagc gatctgtttg
      421 gccagattat taacgcgggc cagggcgaag gccgctatag cgaactgctg gcgattaacc
      481 tgctggaaca gctgctgctg cgccgcatgg aagcgattaa cgaaagcctg catccgccga
      541 tggataaccg cgtgcgcgaa gcgtgccagt atattagcga tcatctggcg gatagcaact
      601 ttgatattgc gagcgtggcg cagcatgtgt gcctgagccc gagccgcctg agccatctgt
      661 ttcgccagca gctgggcatt agcgtgctga gctggcgcga agatcagcgc attagccagg
      721 cgaaactgct gctgagcacc acccgcatgc cgattgcgac cgtgggccgc aacgtgggct
      781 ttgatgatca gctgtatttt agccgcgtgt ttaaaaaatg caccggcgcg agcccgagcg
      841 aatttcgcgc gggctgcgaa gaaaaagtga acgatgtggc ggtgaaactg agcgggtaag
      901 gctgaagagc ctggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
      961 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1021 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa
     1081 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     1141 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     1201 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     1261 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     1321 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     1381 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     1441 ccagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     1501 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     1561 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     1621 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     1681 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     1741 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     1801 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc
     1861 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     1921 acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1981 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2041 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2101 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2161 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2221 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2281 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2341 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2401 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2461 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2521 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2581 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2641 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     2701 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     2761 gtgagcgaga atgctcttct
//
