LOCUS       sequence-242362-        5510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express endogenous biotin ligase in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5510)
  AUTHORS   Lomakin YA, Kaminskaya AN, Stepanov AV, Shmidt AA, Gabibov A, 
            Belogurov AA
  TITLE     Probing surface membrane receptors using engineered bacteriophage 
            bioconjugates.
  JOURNAL   Bioconjug Chem. 2019 Apr 25. doi: 10.1021/acs.bioconjchem.9b00218.
  PUBMED    31021608
REFERENCE   2  (bases 1 to 5510)
  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"; journalName: "Bioconjug 
            Chem. 2019 Apr 25. doi: 10.1021/acs.bioconjchem.9b00218."
COMMENT     Sequence Label: BirA pLEMO
FEATURES             Location/Qualifiers
     source          1..5510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             7..969
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSSEK"
     primer_bind     complement(1025..1043)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1039..1086
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1307..1966)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1881..1900
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1967..2069)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2461..2478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2595..3140)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2675..2694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3471..4319)
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     CDS             complement(4393..5229)
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     promoter        5375..5493
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
ORIGIN
        1 gtcgacatga aggataacac cgtgccactg aaattgattg ccctgttagc gaacggtgaa
       61 tttcactctg gcgagcagtt gggtgaaacg ctgggaatga gccgggcggc tattaataaa
      121 cacattcaga cactgcgtga ctggggcgtt gatgtcttta ccgttccggg taaaggatac
      181 agcctgcctg agcctatcca gttacttaat gctaaacaga tattgggtca gctggatggc
      241 ggtagtgtag ccgtgctgcc agtgattgac tccacgaatc agtaccttct tgatcgtatc
      301 ggagagctta aatcgggcga tgcttgcatt gcagaatacc agcaggctgg ccgtggtcgc
      361 cggggtcgga aatggttttc gccttttggc gcaaacttat atttgtcgat gttctggcgt
      421 ctggaacaag gcccggcggc ggcgattggt ttaagtctgg ttatcggtat cgtgatggcg
      481 gaagtattac gcaagctggg tgcagataaa gttcgtgtta aatggcctaa tgacctctat
      541 ctgcaggatc gcaagctggc aggcattctg gtggagctga ctggcaaaac tggcgatgcg
      601 gcgcaaatag tcattggagc cgggatcaac atggcaatgc gccgtgttga agagagtgtc
      661 gttaatcagg ggtggatcac gctgcaggaa gcggggatca atctcgatcg taatacgttg
      721 gcggccatgc taatacgtga attacgtgct gcgttggaac tcttcgaaca agaaggattg
      781 gcaccttatc tgtcgcgctg ggaaaagctg gataatttta ttaatcgccc agtgaaactt
      841 atcattggtg ataaagaaat atttggcatt tcacgcggaa tagacaaaca gggggcttta
      901 ttacttgagc aggatggaat aataaaaccc tggatgggcg gtgaaatatc cctgcgtagt
      961 tcagaaaagg gatccccgcg ccctcatccg aaagggcgta ttcatatgcc ctaggctgct
     1021 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1081 tttttgctga aacctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat
     1141 aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg
     1201 accgggtcga atttgctttc gaatttctgc cattcatccg cttattatca cttattcagg
     1261 cgtagcaacc aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct
     1321 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     1381 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     1441 atttgcccat agtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     1501 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
     1561 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     1621 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
     1681 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     1741 ccatacggaa ctccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     1801 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     1861 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     1921 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     1981 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     2041 gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg
     2101 gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca
     2161 caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc caacttactg atttagtgta
     2221 tgatggtgtt tttgaggtgc tccagtggct tctgtttcta tcagctgtcc ctcctgttca
     2281 gctactgacg gggtggtgcg taacggcaaa agcaccgccg gacatcagcg ctagcggagt
     2341 gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag
     2401 gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc
     2461 ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga
     2521 acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc
     2581 gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc
     2641 tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc
     2701 ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta
     2761 tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa
     2821 gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta
     2881 tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa
     2941 ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa
     3001 gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga
     3061 gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc
     3121 gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat ttctaggttt
     3181 cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt
     3241 aattctcatg ttagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     3301 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
     3361 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3421 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgcccg ataagcttaa ttaatctttc
     3481 tgcgaattga gatgacgcca ctggctgggc gtcatcccgg tttcccgggt aaacaccacc
     3541 gaaaaatagt tactatcttc aaagccacat tcggtcgaaa tatcactgat taacaggcgg
     3601 ctatgctgga gaagatattg cgcatgacac actctgacct gtcgcagata ttgattgatg
     3661 gtcattccag tctgctggcg aaattgctga cgcaaaacgc gctcactgca cgatgcctca
     3721 tcacaaaatt tatccagcgc aaagggactt ttcaggctag ccgccagccg ggtaatcagc
     3781 ttatccagca acgtttcgct ggatgttggc ggcaacgaat cactggtgta acgatggcga
     3841 ttcagcaaca tcaccaactg cccgaacagc aactcagcca tttcgttagc aaacggcaca
     3901 tgctgactac tttcatgctc aagctgacca ataacctgcc gcgcctgcgc catccccatg
     3961 ctacctaagc gccagtgtgg ttgccctgcg ctggcgttaa atcccggaat cgccccctgc
     4021 cagtcaagat tcagcttcag acgctccggg caataaataa tattctgcaa aaccagatcg
     4081 ttaacggaag cgtaggagtg tttatcatca gcatgaatgt aaaagagatc gccacgggta
     4141 atgcgataag ggcgatcgtt gagtacatgc aggccattac cgcgccagac aatcaccagc
     4201 tcacaaaaat catgtgtatg ttcagcaaag acatcttgcg gataacggtc agccacagcg
     4261 actgcctgct ggtcgctggc aaaaaaatca tctttgagaa gttttaactg atgcgccacc
     4321 gtggctacct cggccagaga acgaagttga ttattcgcaa tatggcgtac aaatacgttg
     4381 agaagattcg cgttattgca gaaagccatc ccgtccctgg cgaatatcac gcggtgacca
     4441 gttaaactct cggcgaaaaa gcgtcgaaaa gtggttactg tcgctgaatc cacagcgata
     4501 ggcgatgtca gtaacgctgg cctcgctgtg gcgtagcaga tgtcgggctt tcatcagtcg
     4561 caggcggttc aggtatcgct gaggcgtcag tcccgtttgc tgcttaagct gccgatgtag
     4621 cgtacgcagt gaaagagaaa attgatccgc cacggcatcc caattcacct catcggcaaa
     4681 atggtcctcc agccaggcca gaagcaagtt gagacgtgat gcgctgtttt ccaggttctc
     4741 ctgcaaactg cttttacgca gcaagagcag taattgcata aacaagatct cgcgactggc
     4801 ggtcgagggt aaatcatttt ccccttcctg ctgttccatc tgtgcaacca gctgtcgcac
     4861 ctgctgcaat acgctgtggt taacgcgcca gtgagacgga tactgcccat ccagctcttg
     4921 tggcagcaac tgattcagcc cggcgagaaa ctgaaatcga tccggcgagc gatacagcac
     4981 attggtcaga cacagattat cggtatgttc atacagatgc cgatcatgat cgcgtacgaa
     5041 acagaccgtg ccaccggtga tggtataggg ctgcccatta aacacatgaa tacccgtgcc
     5101 atgttcgaca atcacaattt catgaaaatc atgatgatgt tcaggaaaat ccgcctgcgg
     5161 gagccggggt tctatcgcca cggacgcgtt accagacgga aaaaaatcca cactatgtaa
     5221 tacggtcata ctggcctcct gatgtcgtca acacggcgaa atagtaatca cgaggtcagg
     5281 ttcttacctt aaattttcga cggaaaacca cgtaaaaaac gtcgattttt caagatacag
     5341 cgtgaatttt caggaaatgc ggtgagcatc acatcaccac aattcagcaa attgtgaaca
     5401 tcatcacgtt catctttccc tggttgccaa tggcccattt tcctgtcagt aacgagaagg
     5461 tcgcgaattc aggcgctttt tagactggtc gtaatgaaat tcaggaggtg
//
