LOCUS       sequence-432940-        7018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flag-tagged biotinylation enzyme for co-expression with 
            bio-proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7018)
  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 7018)
  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: "Genome 
            Res."; date: "2008-04"; volume: "18(4)"; pages: "622-30. Epub 2008 
            Feb 22"
COMMENT     Sequence Label: secreted BirA-Flag
FEATURES             Location/Qualifiers
     source          1..7018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     regulatory      1313..1322
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1379..2338
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             2345..2368
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2398..2453
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2452..2471)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4689..4707)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4775..4879
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4880..4948,4949..5740)
                     /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(5098..5117)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5911..6499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6400..6419
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6653..6670
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6787..6808
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6823..6828,6829..6846,6847..6853)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6861..6877
                     /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     6866..6888
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6885..6901
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6885..6901
                     /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 ccctctagag ccgccaccat
     1321 gggcagacac ctcctgctgc tcctgttcag catcctgtac atgctgctgg cccaggccaa
     1381 ggacaacacc gtgcccctga agctgatcgc cctgctggcc aacggcgagt tccacagcgg
     1441 cgagcagctg ggcgagaccc tgggcatgag cagagccgct atcaacaagc acatccagac
     1501 cctgagagac tggggcgtgg acgtgttcac cgtgcccggc aagggctaca gcctgcccga
     1561 gcccatccag ctgctgaacg ccaagcagat cctgggccag ctggacggcg gaagcgtggc
     1621 cgtgctgccc gtgatcgaca gcaccaacca gtacctgctg gacagaatcg gcgagctgaa
     1681 gagcggcgac gcctgcatcg ccgagtacca gcaggccggc agaggcagaa gaggcagaaa
     1741 gtggttcagc ccctttggcg ccaacctgta cctgagcatg ttctggagac tggagcaggg
     1801 ccccgcagcc gcaatcggcc tgagcctggt gatcggcatc gtgatggccg aggtgctgag
     1861 aaagctgggc gccgacaagg tgagagtgaa gtggcccaac gacctgtacc tgcaggacag
     1921 aaagctggcc ggcatcctgg tggagctgac cggcaagacc ggcgacgccg cacagatcgt
     1981 gatcggcgcc ggcatcaaca tggccatgag aagagtggag gaaagcgtgg tgaaccaggg
     2041 ctggatcacc ctgcaggagg ccggcatcaa cctggacaga aacaccctgg cagccatgct
     2101 gatcagagag ctgagagccg cactggagct gttcgagcag gagggcctgg ctccctacct
     2161 gagcagatgg gagaagctgg acaacttcat caacagaccc gtgaagctga tcatcggcga
     2221 caaggagatc ttcggcatca gcagaggcat cgacaagcag ggcgccctgc tgctggagca
     2281 ggacggcatc atcaagccct ggatgggagg cgagatcagc ctgagaagcg ccgagaaggg
     2341 ccccgactac aaggacgatg atgacaagtg aggatccccc gacctcgacc tctggctaat
     2401 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     2461 ggacatatgg gagggcaaat catttggtcg agatccctcg gagatctcta gctagaggat
     2521 cgatccccgc cccggacgaa ctaaacctga ctacgacatc tctgcccctt cttcgcgggg
     2581 cagtgcatgt aatcccttca gttggttggt acaacttgcc aactgggccc tgttccacat
     2641 gtgacacggg gggggaccaa acacaaaggg gttctctgac tgtagttgac atccttataa
     2701 atggatgtgc acatttgcca acactgagtg gctttcatcc tggagcagac tttgcagtct
     2761 gtggactgca acacaacatt gcctttatgt gtaactcttg gctgaagctc ttacaccaat
     2821 gctgggggac atgtacctcc caggggccca ggaagactac gggaggctac accaacgtca
     2881 atcagagggg cctgtgtagc taccgataag cggaccctca agagggcatt agcaatagtg
     2941 tttataaggc ccccttgtta accctaaacg ggtagcatat gcttcccggg tagtagtata
     3001 tactatccag actaacccta attcaatagc atatgttacc caacgggaag catatgctat
     3061 cgaattaggg ttagtaaaag ggtcctaagg aacagcgata tctcccaccc catgagctgt
     3121 cacggtttta tttacatggg gtcaggattc cacgagggta gtgaaccatt ttagtcacaa
     3181 gggcagtggc tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc gcctgcttct
     3241 tcattctcct tcgtttagct aatagaataa ctgctgagtt gtgaacagta aggtgtatgt
     3301 gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg acggggggtt
     3361 cagtggtggc attgtgctat gacaccaata taaccctcac aaaccccttg ggcaataaat
     3421 actagtgtag gaatgaaaca ttctgaatat ctttaacaat agaaatccat ggggtgggga
     3481 caagccgtaa agactggatg tccatctcac acgaatttat ggctatgggc aacacataat
     3541 cctagtgcaa tatgatactg gggttattaa gatgtgtccc aggcagggac caagacaggt
     3601 gaaccatgtt gttacactct atttgtaaca aggggaaaga gagtggacgc cgacagcagc
     3661 ggactccact ggttgtctct aacacccccg aaaattaaac ggggctccac gccaatgggg
     3721 cccataaaca aagacaagtg gccactcttt tttttgaaat tgtggagtgg gggcacgcgt
     3781 cagcccccac acgccgccct gcggttttgg actgtaaaat aagggtgtaa taacttggct
     3841 gattgtaacc ccgctaacca ctgcggtcaa accacttgcc cacaaaacca ctaatggcac
     3901 cccggggaat acctgcataa gtaggtgggc gggccaagat aggggcgcga ttgctgcgat
     3961 ctggaggaca aattacacac acttgcgcct gagcgccaag cacagggttg ttggtcctca
     4021 tattcacgag gtcgctgaga gcacggtggg ctaatgttgc catgggtagc atatactacc
     4081 caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa
     4141 tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa
     4201 tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa
     4261 tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctacccaga
     4321 tagagattag ggtagtatat gctatcctaa tttatatctg ggtagcatat actacccaaa
     4381 tatctggata gcatatgcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     4441 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     4501 tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta
     4561 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
     4621 tatccgggta gcatatgcta tcctcatgat aagctgtcaa acatgagaat tttcttgaag
     4681 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4741 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4801 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4861 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4921 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4981 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5041 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5101 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     5161 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5221 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5281 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5341 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5401 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5461 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5521 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5581 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5641 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5701 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5761 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5821 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5881 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5941 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6001 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6061 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6121 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6181 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6241 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6301 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6361 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6421 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6481 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6541 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6601 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6661 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6721 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6781 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6841 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6901 ccatgattac gccaagctct agctagaggt cgaccaattc tcatgtttga cagcttatca
     6961 tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatgga
//
