LOCUS       40924_2179        6386 bp DNA     circular SYN 13-MAY-2021
DEFINITION  To fuse your protein of interest to the C-terminus of BioID2 and use
            in proximity-dependent biotin identification (BioID); MCS and a 25nm
            GS linker are present downstream of BioID2 and myc tags..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6386)
  AUTHORS   Kim DI, Jensen SC, Noble KA, Kc B, Roux KH, Motamedchaboki K, Roux 
            KJ
  TITLE     An improved smaller biotin ligase for BioID proximity labeling.
  JOURNAL   Mol Biol Cell. 2016 Feb 24. pii: mbc.E15-12-0844.
  PUBMED    26912792
REFERENCE   2  (bases 1 to 6386)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6386)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Biol 
            Cell. 2016 Feb 24. pii: mbc.E15-12-0844."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    6..230
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      276..704
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        718..1047
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             1114..1905
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    2082..2215
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2252..2268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2276..2292)
                     /label=lac operator
                     /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)."
     promoter        complement(2300..2330)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2345..2366)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(2483..2500)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2654..3242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3416..4273)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4274..4378)
                     /label=AmpR promoter
     primer_bind     complement(4453..4472)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     enhancer        4644..5023
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5024..5227
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        5272..5290
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5308..5317
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5317..5346
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5350..6045
                     /codon_start=1
                     /label=BioID2
                     /note="promiscuous R40G mutant of a biotin protein ligase
                     from Aquifex aeolicus (Kim et al., 2016)"
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
ORIGIN
        1 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
       61 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
      121 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
      181 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
      241 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
      301 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
      361 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
      421 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
      481 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
      541 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
      601 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
      661 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
      721 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
      781 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
      841 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
      901 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
      961 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     1021 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     1081 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     1141 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     1201 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     1261 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     1321 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     1381 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     1441 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     1501 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     1561 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     1621 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     1681 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     1741 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     1801 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     1861 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     1921 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     1981 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     2041 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     2101 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2161 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     2221 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2281 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2341 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2401 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2461 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2521 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2581 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2641 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2701 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2761 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2821 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2881 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2941 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3001 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3061 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3121 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3181 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3241 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3301 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3361 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3421 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3481 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3541 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3601 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3661 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3721 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3781 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3841 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3901 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3961 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4021 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4081 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4141 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4201 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4261 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4321 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4381 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     4441 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     4501 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     4561 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     4621 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     4681 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     4741 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     4801 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     4861 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     4921 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     4981 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     5041 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     5101 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     5161 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     5221 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
     5281 tcactatagg gagacccaag ctggctagcc accatggaac aaaaactcat ctcagaagag
     5341 gatctcgact tcaagaacct gatctggctg aaggaggtgg acagcaccca ggagagactg
     5401 aaggagtgga acgtgagcta cggcaccgcc ctggtggccg acagacagac caagggcaga
     5461 ggcggcctgg gcagaaagtg gctgagccag gagggcggcc tgtacttcag cttcctgctg
     5521 aaccccaagg agttcgagaa cctgctgcag ctgcccctgg tgctgggcct gagcgtgagc
     5581 gaggccctgg aggagatcac cgagatcccc ttcagcctga agtggcccaa cgacgtgtac
     5641 ttccaggaga agaaggtgag cggcgtgctg tgcgagctga gcaaggacaa gctgatcgtg
     5701 ggcatcggca tcaacgtgaa ccagagagag atccccgagg agatcaagga cagagccacc
     5761 accctgtacg agatcaccgg caaggactgg gacagaaagg aggtgctgct gaaggtgctg
     5821 aagagaatca gcgagaacct gaagaagttc aaggagaaga gcttcaagga gttcaagggc
     5881 aagatcgaga gcaagatgct gtacctgggc gaggaggtga agctgctggg cgagggcaag
     5941 atcaccggca agctggtggg cctgagcgag aagggcggcg ccctgatcct gaccgaggag
     6001 ggcatcaagg agatcctgag cggcgagttc agcctgagaa gaagcaccgg tttttcggaa
     6061 ttcggatccg gtggaggcgg gtctggaggc gggggtagtg gcgggggtgg aagcgggggt
     6121 ggaggcgggt cgggtggcgg aggtagcgga ggcggtggaa gtggaggcgg tggaagtggt
     6181 ggaggaggtt ctggtggcgg aggtagcggt ggcggaggta gcggaggcgg tggaagtggt
     6241 ggcggaggta gcggaggcgg tggatcggcg cgccgtttca agaacctcga gcggccgcca
     6301 ctgtgctgga tatctgcaga attccaccac actggactag tggatccgag ctcggtacca
     6361 agcttaagtt taaaccgctg atcagc
//
