LOCUS       Exported                7001 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Flag-tagged biotinylation enzyme for co-expression with 
            bio-proteins.
ACCESSION   .
VERSION     .
KEYWORDS    secreted BirA-Flag
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7001)
  AUTHORS   Bushell KM, Sollner C, Schuster-Boeckler B, Bateman A, Wright GJ
  TITLE     Large-scale screening for novel low-affinity extracellular protein 
            interactions.
  JOURNAL   Genome Res. 2008 Apr;18(4):622-30. Epub 2008 Feb 22.
  PUBMED    18296487
REFERENCE   2  (bases 1 to 7001)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7001
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        154..533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        534..737
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     688..708
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     734..758
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1239..1258
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1362..2321
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             2328..2351
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2381..2436
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2435..2454)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4672..4690)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4758..4862
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4863..5723
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5081..5100)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5894..6482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6383..6402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6636..6653
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6770..6791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6806..6836
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6844..6860
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6849..6871
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6868..6884
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6868..6884
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
       61 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
      121 atattggctc atgtccaata tgaccgccat gttgacattg attattgact agttattaat
      181 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      241 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      301 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      361 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
      421 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
      481 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      541 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      601 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      661 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
      721 tctatataag cagagctcgt ttagtgaacc gtcagatcct cactctcttc cgcatcgctg
      781 tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt
      841 actcttggat cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc
      901 gagtccgcat cgaccggatc ggaaaacctc tcgagaaagg cgtctaacca gtcacagtcg
      961 caaggtaggc tgagcaccgt ggcgggcggc agcgggtggc ggtcggggtt gtttctggcg
     1021 gaggtgctgc tgatgatgta attaaagtag gcggtcttga gacggcggat ggtcgaggtg
     1081 aggtgtggca ggcttgagat ccagctgttg gggtgagtac tccctctcaa aagcgggcat
     1141 tacttctgcg ctaagattgt cagtttccaa aaacgaggag gatttgatat tcacctggcc
     1201 cgatctggcc atacacttga gtgacaatga catccacttt gcctttctct ccacaggtgt
     1261 ccactcccag gtccaagttt aaacggatct ctagcgaatt catgggcaga cacctcctgc
     1321 tgctcctgtt cagcatcctg tacatgctgc tggcccaggc caaggacaac accgtgcccc
     1381 tgaagctgat cgccctgctg gccaacggcg agttccacag cggcgagcag ctgggcgaga
     1441 ccctgggcat gagcagagcc gctatcaaca agcacatcca gaccctgaga gactggggcg
     1501 tggacgtgtt caccgtgccc ggcaagggct acagcctgcc cgagcccatc cagctgctga
     1561 acgccaagca gatcctgggc cagctggacg gcggaagcgt ggccgtgctg cccgtgatcg
     1621 acagcaccaa ccagtacctg ctggacagaa tcggcgagct gaagagcggc gacgcctgca
     1681 tcgccgagta ccagcaggcc ggcagaggca gaagaggcag aaagtggttc agcccctttg
     1741 gcgccaacct gtacctgagc atgttctgga gactggagca gggccccgca gccgcaatcg
     1801 gcctgagcct ggtgatcggc atcgtgatgg ccgaggtgct gagaaagctg ggcgccgaca
     1861 aggtgagagt gaagtggccc aacgacctgt acctgcagga cagaaagctg gccggcatcc
     1921 tggtggagct gaccggcaag accggcgacg ccgcacagat cgtgatcggc gccggcatca
     1981 acatggccat gagaagagtg gaggaaagcg tggtgaacca gggctggatc accctgcagg
     2041 aggccggcat caacctggac agaaacaccc tggcagccat gctgatcaga gagctgagag
     2101 ccgcactgga gctgttcgag caggagggcc tggctcccta cctgagcaga tgggagaagc
     2161 tggacaactt catcaacaga cccgtgaagc tgatcatcgg cgacaaggag atcttcggca
     2221 tcagcagagg catcgacaag cagggcgccc tgctgctgga gcaggacggc atcatcaagc
     2281 cctggatggg aggcgagatc agcctgagaa gcgccgagaa gggccccgac tacaaggacg
     2341 atgatgacaa gtgaggatcc cccgacctcg acctctggct aataaaggaa atttattttc
     2401 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     2461 aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc cgccccggac
     2521 gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca tgtaatccct
     2581 tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac ggggggggac
     2641 caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg tgcacatttg
     2701 ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact gcaacacaac
     2761 attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg gacatgtacc
     2821 tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag gggcctgtgt
     2881 agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa ggcccccttg
     2941 ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc cagactaacc
     3001 ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta gggttagtaa
     3061 aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt ttatttacat
     3121 ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt ggctgaagat
     3181 caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct ccttcgttta
     3241 gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc tcgaaaacaa
     3301 ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt ggcattgtgc
     3361 tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg taggaatgaa
     3421 acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg taaagactgg
     3481 atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg caatatgata
     3541 ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat gttgttacac
     3601 tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc actggttgtc
     3661 tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa acaaagacaa
     3721 gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc cacacgccgc
     3781 cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta accccgctaa
     3841 ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg aatacctgca
     3901 taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg acaaattaca
     3961 cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac gaggtcgctg
     4021 agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat ctggatagca
     4081 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4141 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     4201 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4261 tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat tagggtagta
     4321 tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg atagcatatg
     4381 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagcatagg
     4441 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     4501 ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     4561 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg
     4621 ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag ggcctcgtga
     4681 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4741 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4801 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4861 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4921 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4981 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5041 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5101 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5161 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5221 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5281 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5341 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5401 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5461 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5521 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5581 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5641 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5701 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5761 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5821 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5881 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5941 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6001 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6061 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6121 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6181 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6241 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6301 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6361 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6421 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6481 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6541 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6601 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6661 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6721 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     6781 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     6841 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     6901 tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga tccgggcaac
     6961 gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg a
//
