LOCUS       sequence-484661-        3215 bp    DNA     circular SYN 26-AUG-2025
DEFINITION  Contains an AraC R5 Variant-LOV Chimera 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 3215)
  TITLE     Phage-Assisted Evolution of Allosteric Protein Switches
REFERENCE   2  (bases 1 to 3215)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 26, 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_R5_S170_LOV
FEATURES             Location/Qualifiers
     source          1..3215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     227..244
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1763..2578)
                     /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(1763..2578)
                     /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     2486..2505
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2570..2589
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(2700..3215,1..73)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3189..3208
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
       61 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      121 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      181 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
      241 cgagaatgct cttctagtaa ggaggaaaaa tgtccgagga agcgcagaac gatccgctgc
      301 tgccgggcta tagctttaac gcgcatctgg tggcgggcct gaccccgatt gaagcgaacg
      361 gctatctgga tttttttatt gatcgcccgc tgggcatgaa aggctatatt ctgaacctga
      421 ccattcgcgg ccagggcgtg gtgaaaaacc agggccgcga atttgtgtgc cgcccgggcg
      481 atattctgct gtttccgccg ggcgaaattc atcattatgg ccgccatccg gaagcgcgcg
      541 aatggtatca tcagtgggtg tattttcgcc cgcgcgcgta ttggcatgaa tggctgaact
      601 ggccgagcat ttttgcgaac accggctttt ttcgcccgga tgaagcgcat cagccgcatt
      661 ttagcgatct gtttggccag attattaacg cgggccaggg cgaaggccgc tatagcgaac
      721 tgctggcgat taacctgata gaacagctgc tgctgcgccg catggaagcg attaacgaaa
      781 gcagcggttt agccacaacg ctggaacgca ttgaaaagaa tttcgtaatc acagacccgc
      841 gccttcccga caatccaatt atttttgcgt ccgatagctt cctgcaatta accgaataca
      901 gccgcgaaga aattctgggt cgtaattgtc gcttccttca ggggccagag actgaccgtg
      961 ctacggtacg caaaatccgc gacgcaatcg acaatcaaac ggaagtcacg gttcagttga
     1021 ttaactatac gaagagcgga aaaaaattct ggaatttatt tcacttgcag cctatgcgtg
     1081 accagaaggg cgatgtccag tatttcattg gcgttcagct tgatggtacc gagcatgttc
     1141 gcgatgctgc ggagcgtgaa ggtgtaatgt taattaaaaa gactgctgaa aacattgatg
     1201 aggcggccaa agggagcctg catccgccga tggataaccg cgtgcgcgaa gcgtgccagt
     1261 atattagcga tcatctggcg gatagcaact ttgatattgc gagcgtggcg cagcatgtgt
     1321 gcctgagccc gagccgcctg agccatctgt ttcgccagca gctgggcatt agcgtgctga
     1381 gctggcgcga agatcagcgc attagccagg cgaaactgct gctgagcacc acccgcatgc
     1441 cgattgcgac cgtgggccac aacgtgggct ttgatgatca gctgtatttt agccgcgtgt
     1501 ttaaaaaatg caccggcgcg agcccgagcg aatttcgcgc gggctgcgaa gaaaaagtga
     1561 acgatgtggc ggtgaaactg agcgggtaag gctgaagagc ctggttaaaa aatgagctga
     1621 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     1681 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1741 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
     1801 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     1861 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     1921 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     1981 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     2041 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     2101 aaccgttatt cattcgtgat tgcgcctgag ccagacgaaa tacgcgatcg ctgttaaaag
     2161 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     2221 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     2281 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     2341 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     2401 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     2461 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     2521 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     2581 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct
     2641 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2701 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2761 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2821 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2881 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2941 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3001 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3061 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3121 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3181 cttccagggg gaaacgcctg gtatctttat agtcc
//
