LOCUS       Exported                5508 bp ds-DNA     circular SYN 20-MAR-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    BirA pLEMO
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5508)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 1, 2018 from SnapGene 4.2.6
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5508
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             7..969
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSSEK"
     terminator      1039..1086
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1306..1965)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1966..2068)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(2594..3138)
                     /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."
     CDS             complement(3469..4317)
                     /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(4391..5227)
                     /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        5373..5491
                     /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 tcagaaaaag 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 cgtagcacca ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg
     1321 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     1381 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     1441 tttgcccata gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     1501 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     1561 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     1621 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     1681 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     1741 catacggaac tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     1801 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     1861 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     1921 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     1981 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     2041 aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg
     2101 cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac
     2161 aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga tttagtgtat
     2221 gatggtgttt ttgaggtgct ccagtggctt ctgtttctat cagctgtccc tcctgttcag
     2281 ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg
     2341 tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg
     2401 agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc
     2461 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa
     2521 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg
     2581 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct
     2641 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt cccctggcgg
     2701 ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg
     2761 gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc
     2821 tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc
     2881 gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt
     2941 gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt
     3001 tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga
     3061 accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc
     3121 agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt ctaggtttca
     3181 gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa
     3241 ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     3301 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     3361 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3421 aacgcgcggg gagaggcggt ttgcgtattg ggcgcccgat aagcttaatt aatctttctg
     3481 cgaattgaga tgacgccact ggctgggcgt catcccggtt tcccgggtaa acaccaccga
     3541 aaaatagtta ctatcttcaa agccacattc ggtcgaaata tcactgatta acaggcggct
     3601 atgctggaga agatattgcg catgacacac tctgacctgt cgcagatatt gattgatggt
     3661 cattccagtc tgctggcgaa attgctgacg caaaacgcgc tcactgcacg atgcctcatc
     3721 acaaaattta tccagcgcaa agggactttt caggctagcc gccagccggg taatcagctt
     3781 atccagcaac gtttcgctgg atgttggcgg caacgaatca ctggtgtaac gatggcgatt
     3841 cagcaacatc accaactgcc cgaacagcaa ctcagccatt tcgttagcaa acggcacatg
     3901 ctgactactt tcatgctcaa gctgaccgat aacctgccgc gcctgcgcca tccccatgct
     3961 acctaagcgc cagtgtggtt gccctgcgct ggcgttaaat cccggaatcg ccccctgcca
     4021 gtcaagattc agcttcagac gctccgggca ataaataata ttctgcaaaa ccagatcgtt
     4081 aacggaagcg taggagtgtt tatcgtcagc atgaatgtaa aagagatcgc cacgggtaat
     4141 gcgataaggg cgatcgttga gtacatgcag gccattaccg cgccagacaa tcaccagctc
     4201 acaaaaatca tgtgtatgtt cagcaaagac atcttgcgga taacggtcag ccacagcgac
     4261 tgcctgctgg tcgctggcaa aaaaatcatc tttgagaagt tttaactgat gcgccaccgt
     4321 ggctacctcg gccagagaac gaagttgatt attcgcaata tggcgtacaa atacgttgag
     4381 aagattcgcg ttattgcaga aagccatccc gtccctggcg aatatcacgc ggtgaccagt
     4441 taaactctcg gcgaaaaagc gtcgaaaagt ggttactgtc gctgaatcca cagcgatagg
     4501 cgatgtcagt aacgctggcc tcgctgtggc gtagcagatg tcgggctttc atcagtcgca
     4561 ggcggttcag gtatcgctga ggcgtcagtc ccgtttgctg cttaagctgc cgatgtagcg
     4621 tacgcagtga aagagaaaat tgatccgcca cggcatccca attcacctca tcggcaaaat
     4681 ggtcctccag ccaggccaga agcaagttga gacgtgatgc gctgttttcc aggttctcct
     4741 gcaaactgct tttacgcagc aagagcagta attgcataaa caagatctcg cgactggcgg
     4801 tcgagggtaa atcattttcc ccttcctgct gttccatctg tgcaaccagc tgtcgcacct
     4861 gctgcaatac gctgtggtta acgcgccagt gagacggata ctgcccatcc agctcttgtg
     4921 gcagcaactg attcagcccg gcgagaaact gaaatcgatc cggcgagcga tacagcacat
     4981 tggtcagaca cagattatcg gtatgttcat acagatgccg atcatgatcg cgtacgaaac
     5041 agaccgtgcc accggtgatg gtatagggct gcccattaaa cacatgaata cccgtgccat
     5101 gttcgacaat cacaatttca tgaaaatcat gatgatgttc aggaaaatcc gcctgcggga
     5161 gccggggttc tatcgccacg gacgcgttac cagacggaaa aaaatccaca ctatgtaata
     5221 cggtcatact ggcctcctga tgtcgtcaac acggcgaaat agtaatcacg aggtcaggtt
     5281 cttaccttaa attttcgacg gaaaaccacg taaaaaacgt cgatttttca agatacagcg
     5341 tgaattttca ggaaatgcgg tgagcatcac atcaccacaa ttcagcaaat tgtgaacatc
     5401 atcacgttca tctttccctg gttgccaatg gcccattttc ctgtcagtaa cgagaaggtc
     5461 gcgaattcag gcgcttttta gactggtcgt aatgaaattc aggaggtg
//
