LOCUS       40924_6272        6573 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBG1805, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6573)
  AUTHORS   O'Connor CM, Denninger AR.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-AUG-2011) Biology, Boston College, 140 Commonwealth 
            Ave., Chestnut Hill, MA 02467, USA
REFERENCE   2  (bases 1 to 6573)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6573)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (07-AUG-2011) Biology, Boston College, 140 Commonwealth Ave., 
            Chestnut Hill, MA 02467, USA"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(2..26)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             40..57
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             70..96
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             100..123
                     /codon_start=1
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
                     /translation="LEVLFQGP"
     CDS             136..309
                     /codon_start=1
                     /label=ProtA
                     /note="IgG-binding unit of Staphylococcus aureus protein A"
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             313..483
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     promoter        complement(529..547)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(557..573)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      714..1169
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(1303..2103)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(2104..2319)
                     /label=URA3 promoter
     rep_origin      2588..3930
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3956..4060
                     /label=AmpR promoter
     CDS             4061..4918
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5030..5618
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5906..5927
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5942..5972
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5980..5996
                     /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)."
     primer_bind     6004..6020
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6041..6059
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6090..6531
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    6544..6568
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 aacccagctt tcttgtacaa agtggttgat gtgtctagac atcatcatca ccatcatggt
       61 agaatctttt atccatacga tgttcctgat tatgctggtt tagaagttct ttttcaaggt
      121 cctggaccat cggccgtgga caacaaattc aacaaagaac aacaaaacgc gttctatgag
      181 atcttacatt tacctaactt aaacgaagaa caacgaaacg ccttcatcca aagtttaaaa
      241 gatgacccaa gccaaagcgc taacctttta gcagaagcta aaaagctaaa tgatgctcag
      301 gcgccgaaag tagacaacaa attcaacaaa gaacaacaaa acgcgttcta tgagatctta
      361 catttaccta acttaaacga agaacaacga aacgccttca tccaaagttt aaaagatgac
      421 ccaagccaaa gcgctaacct tttagcagaa gctaaaaagc taaatgatgc tcaggcgccg
      481 aaagtagacc cgaatcatca gtgaggcgcg ccggggtacc caattcgccc tatagtgagt
      541 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
      601 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
      661 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
      721 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
      781 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
      841 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
      901 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
      961 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1021 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1081 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1141 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     1201 tgcggtattt cacaccgcat agggtaataa ctgatataat taaattgaag ctctaatttg
     1261 tgagtttagt atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct
     1321 tctcaaatat gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct
     1381 tccctttgca aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc
     1441 atccacggtt ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg
     1501 tgtcataatc aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa
     1561 gccgataaca aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc
     1621 agtagatagg gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt
     1681 tgttacttct tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg
     1741 tgcattcgta atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt
     1801 gactgtatta ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc
     1861 ggataatgcc tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac
     1921 atgtgttttt agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt
     1981 ggtggtacga acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa
     2041 tagcttggca gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga
     2101 catgatttat cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta agaatactgg
     2161 gcaatttcat gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc
     2221 cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc aaggaaaccg
     2281 aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaat tgaaaagctg tggtatggtg
     2341 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     2401 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     2461 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     2521 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     2581 ttagtatgat ccaatatcaa aggaaatgat agcattgaag gatgagacta atccaattga
     2641 ggagtggcag catatagaac agctaaaggg tagtgctgaa ggaagcatac gataccccgc
     2701 atggaatggg ataatatcac aggaggtact agactacctt tcatcctaca taaatagacg
     2761 catataagta cgcatttaag cataaacacg cactatgccg ttcttctcat gtatatatat
     2821 atacaggcaa cacgcagata taggtgcgac gtgaacagtg agctgtatgt gcgcagctcg
     2881 cgttgcattt tcggaagcgc tcgttttcgg aaacgctttg aagttcctat tccgaagttc
     2941 ctattctcta gaaagtatag gaacttcaga gcgcttttga aaaccaaaag cgctctgaag
     3001 acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa atattgcgaa
     3061 taccgcttcc acaaacattg ctcaaaagta tctctttgct atatatctct gtgctatatc
     3121 cctatataac ctacccatcc acctttcgct ccttgaactt gcatctaaac tcgacctcta
     3181 cattttttat gtttatctct agtattactc tttagacaaa aaaattgtag taagaactat
     3241 tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt agtatataga
     3301 gacaaaatag aagaaaccgt tcataatttt ctgaccaatg aagaatcatc aacgctatca
     3361 ctttctgttc acaaagtatg cgcaatccac atcggtatag aatataatcg gggatgcctt
     3421 tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg aagtggagtc
     3481 aggctttttt tatggaagag aaaatagaca ccaaagtagc cttcttctaa ccttaacgga
     3541 cctacagtgc aaaaagttat caagagactg cattatagag cgcacaaagg agaaaaaaag
     3601 taatctaaga tgctttgtta gaaaaatagc gctctcggga tgcatttttg tagaacaaaa
     3661 aagaagtata gattctttgt tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa
     3721 aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta
     3781 caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc atttctgttc
     3841 tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg
     3901 ttctacaaaa tgaagcacag atgcttcgtt caggtggcac ttttcgggga aatgtcgcgg
     3961 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4021 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4081 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4141 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4201 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4261 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4321 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4381 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4441 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4501 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4561 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4621 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4681 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4741 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4801 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4861 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4921 actgtcagac caagtttact catatatact ttctcatgac caaaatccct taacgtgagt
     4981 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5041 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5101 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5161 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5221 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5281 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5341 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5401 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5461 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5521 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5581 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5641 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5701 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     5761 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     5821 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     5881 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     5941 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     6001 acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg
     6061 aacaaaagct ggagctccac cgcggggaac ggattagaag ccgccgagcg ggcgacagcc
     6121 ctccgacgga agactctcct ccgtgcgtcc tcgtcttcac cggtcgcgtt cctgaaacgc
     6181 agatgtgcct cgcgccgcac tgctccgaac aataaagatt ctacaatact agcttttatg
     6241 gttatgaaga ggaaaaattg gcagtaacct ggccccacaa accttcaaat taacgaatca
     6301 aattaacaac cataggatga taatgcgatt agttttttag ccttatttct ggggtaatta
     6361 atcagcgaag cgatgatttt tgatctatta acagatatat aaatggaaaa gctgcataac
     6421 cactttaact aatactttca acattttcag tttgtattac ttcttattca aatgtcataa
     6481 aagtatcaac aaaaaattgt taatatacct ctatacttta acgtcaagga gaaggaatta
     6541 tcaacaagtt tgtacaaaaa agcaggctac aaa
//
