LOCUS       40924_9391        4482 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Co-expression vector pCDF-BIRA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4482)
  AUTHORS   Keates T, Cooper CD, Savitsky P, Allerston CK, Phillips C, 
            Hammarstrom M, Daga N, Berridge G, Mahajan P, Burgess-Brown NA, 
            Muller S, Graslund S, Gileadi O.
  TITLE     Expressing the human proteome for affinity proteomics: optimising 
            expression of soluble protein domains and in vivo biotinylation
  JOURNAL   N Biotechnol 29 (5), 515-525 (2012)
  PUBMED    22027370
REFERENCE   2  (bases 1 to 4482)
  AUTHORS   Savitsky P, Gileadi O.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAY-2011) Structural Genomics Consortium, University 
            of Oxford, Old Road Campus Research Building, Roosevelt Drive, 
            Oxford, Oxfordshire OX3 7DQ, UK
REFERENCE   3  (bases 1 to 4482)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4482)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "N 
            Biotechnol"; date: "2012"; volume: "29"; issue: "5"; pages: 
            "515-525"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-MAY-2011) Structural Genomics Consortium, University of Oxford, 
            Old Road Campus Research Building, Roosevelt Drive, Oxford, 
            Oxfordshire OX3 7DQ, UK"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        84..102
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    103..127
                     /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)."
     RBS             142..164
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             174..1136
                     /codon_start=1
                     /label=BirA
                     /note="E. coli biotin protein ligase"
                     /translation="VKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             1167..1211
                     /codon_start=1
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /translation="KETAAAKFERQHMDS"
     terminator      1263..1310
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1484..2272)
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2273..2364)
                     /label=AmpR promoter
     rep_origin      complement(2412..3150)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     protein_bind    complement(3326..3347)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3363..4442)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(join(4443..4482,1..38))
                     /label=lacI promoter
ORIGIN
        1 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
       61 atgcgactcc tgcattagga aattaatacg actcactata ggggaattgt gagcggataa
      121 caattcccct gtagaaataa ttttgtttaa ctttaataag gagatatacc atggtgaagg
      181 ataacaccgt gccactgaaa ttgattgccc tgttagcgaa cggtgaattt cactctggcg
      241 agcagttggg tgaaacgctg ggaatgagcc gggcggctat taataaacac attcagacac
      301 tgcgtgactg gggcgttgat gtctttaccg ttccgggtaa aggatacagc ctgcctgagc
      361 ctatccagtt acttaatgct aaacagatat tgggtcagct ggatggcggt agtgtagccg
      421 tgctgccagt gattgactcc acgaatcagt accttcttga tcgtatcgga gagcttaaat
      481 cgggcgatgc ttgcattgca gaataccagc aggctggccg tggtcgccgg ggtcggaaat
      541 ggttttcgcc ttttggcgca aacttatatt tgtcgatgtt ctggcgtctg gaacaaggcc
      601 cggcggcggc gattggttta agtctggtta tcggtatcgt gatggcggaa gtattacgca
      661 agctgggtgc agataaagtt cgtgttaaat ggcctaatga cctctatctg caggatcgca
      721 agctggcagg cattctggtg gagctgactg gcaaaactgg cgatgcggcg caaatagtca
      781 ttggagccgg gatcaacatg gcaatgcgcc gtgttgaaga gagtgtcgtt aatcaggggt
      841 ggatcacgct gcaggaagcg gggatcaatc tcgatcgtaa tacgttggcg gccatgctaa
      901 tacgtgaatt acgtgctgcg ttggaactct tcgaacaaga aggattggca ccttatctgt
      961 cgcgctggga aaagctggat aattttatta atcgcccagt gaaacttatc attggtgata
     1021 aagaaatatt tggcatttca cgcggaatag acaaacaggg ggctttatta cttgagcagg
     1081 atggaataat aaaaccctgg atgggcggtg aaatatccct gcgtagtgca gaaaaatgac
     1141 agtaaaggtg gatactcgag tctggtaaag aaaccgctgc tgcgaaattt gaacgccagc
     1201 acatggactc gtctactagc gcagcttaat taacctaggc tgctgccacc gctgagcaat
     1261 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaacctc
     1321 aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc
     1381 aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg tcatcgtggc
     1441 cggatcttgc ggcccctcgg cttgaacgaa ttgttagaca ttatttgccg actaccttgg
     1501 tgatctcgcc tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaagc
     1561 gatcttcttc ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg
     1621 ccggcaggcg ctccattgcc cagtcggcag cgacatcctt cggcgcgatt ttgccggtta
     1681 ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt
     1741 cgggcggcga gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag
     1801 gaaccggatc aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg
     1861 cttttgtcag caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa
     1921 gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg
     1981 gaatgatgtc gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc
     2041 caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa
     2101 gccttacggt caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca
     2161 ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc
     2221 caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc atactcttcc
     2281 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2341 aatgtattta gaaaaataaa caaatagcta gctcactcgg tcgctacgct ccgggcgtga
     2401 gactgcggcg ggcgctgcgg acacatacaa agttacccac agattccgtg gataagcagg
     2461 ggactaacat gtgaggcaaa acagcagggc cgcgccggtg gcgtttttcc ataggctccg
     2521 ccctcctgcc agagttcaca taaacagacg cttttccggt gcatctgtgg gagccgtgag
     2581 gctcaaccat gaatctgaca gtacgggcga aacccgacag gacttaaaga tccccaccgt
     2641 ttccggcggg tcgctccctc ttgcgctctc ctgttccgac cctgccgttt accggatacc
     2701 tgttccgcct ttctccctta cgggaagtgt ggcgctttct catagctcac acactggtat
     2761 ctcggctcgg tgtaggtcgt tcgctccaag ctgggctgta agcaagaact ccccgttcag
     2821 cccgactgct gcgccttatc cggtaactgt tcacttgagt ccaacccgga aaagcacggt
     2881 aaaacgccac tggcagcagc cattggtaac tgggagttcg cagaggattt gtttagctaa
     2941 acacgcggtt gctcttgaag tgtgcgccaa agtccggcta cactggaagg acagatttgg
     3001 ttgctgtgct ctgcgaaagc cagttaccac ggttaagcag ttccccaact gacttaacct
     3061 tcgatcaaac cacctcccca ggtggttttt tcgtttacag ggcaaaagat tacgcgcaga
     3121 aaaaaaggat ctcaagaaga tcctttgatc ttttctactg aaccgctcta gatttcagtg
     3181 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc
     3241 tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     3301 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     3361 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3421 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     3481 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     3541 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     3601 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     3661 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     3721 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     3781 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     3841 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     3901 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     3961 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     4021 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     4081 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     4141 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     4201 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     4261 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     4321 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     4381 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     4441 acattcacca ccctgaattg actctcttcc gggcgctatc at
//
