LOCUS       sequence-181410-        6572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For E.coli expression of 6xHis-Tagged BOA (Burkholderia oklahomensis
            agglutinin).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6572)
  AUTHORS   Whitley MJ, Furey W, Kollipara S, Gronenborn AM
  TITLE     Burkholderia oklahomensis agglutinin is a canonical two-domain 
            OAA-family lectin: structures, carbohydrate binding and anti-HIV 
            activity.
  JOURNAL   FEBS J. 2013 May;280(9):2056-67. doi: 10.1111/febs.12229. Epub 2013 
            Apr 2.
  PUBMED    23480609
REFERENCE   2  (bases 1 to 6572)
  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"; doi: 
            "10.1111/febs.12229"; journalName: "FEBS J"; date: "2013-05"; 
            volume: "280"; issue: "9"; pages: "2056-67"
COMMENT     Sequence Label: pET15b/BOA
FEATURES             Location/Qualifiers
     source          1..6572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             163..183
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(1074..1092)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1088..1135
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1310..1315,1316..1332,1333..1338))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1365..1383)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1451..1555
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1556..1624,1625..2416)
                     /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(1774..1793)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2587..3175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3076..3095
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3329..3346
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3361..3500
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3426..3445)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3586..3608
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3602..3793)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6148..6167
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6187..6264)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6470..6489
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccacaag actggccata tggaaaacct gtatttccag
      181 agtgtcgaca tgaccaaaac ttctaaaggt tttaataact tacagcacgt gcagaaccaa
      241 tggggtggct cctccgcacc gtggcacgaa ggcggtatgt gggtcctggg ttgccgttcc
      301 ggccagaacg ttgtcgccct gaacattaaa tcgggggatg gtggtcgcac cctgacaggt
      361 acgatgacct acgttggtga aggtccgatc ggcttccgtg ctaccctgac ccagtctaac
      421 acctacgccg tggaaaacca gtggggcggc tcttcagctc cgtggcaccc gggcggcacc
      481 tgggttatcg gttgccgcgt gaaccagcaa gttgttgctc tggatattga aagcggcgat
      541 cagggcgcga ccctggcagg taccatgacc tacgctggcg aaggtccgat cggctttaaa
      601 agccagcagg cggatggcgg tgtgtacgcg gttgaaaacc agtggggtgg tagcagcgcg
      661 ccgtggcaca acggtggcgt gtgggtaatc ggtgcacgtg accaggcggt agtagcagtt
      721 tccatcggtt ccaccgatag cggcaaaacc ctgaacggta acatgaccta tgctggtgag
      781 ggtccgatcg gtttcaaagg taatagtgtg gccggtaaca actacgcggt ggaaaaccag
      841 tggggtggta cctcggcgcc gtggcacccg ggcggcattt ggctgctggg ttgccgctct
      901 ggtcagaacg tcgtagaact gtatatcact tctggggata acggtaacac ttttcacgga
      961 tctatgacct acagcggaga aggtccgatc ggtttccgcg cgatggctct gccgcagtag
     1021 taaggatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1081 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1141 aggaggaact atatccggat atcccgcaag aggcccggca gtaccggcat aaccaagcct
     1201 atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc
     1261 atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta
     1321 tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg tgatacgcct
     1381 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     1441 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1501 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1561 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1621 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1681 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1741 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     1801 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1861 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1921 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1981 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2041 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     2101 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2161 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2221 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2281 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2341 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2401 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2461 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2521 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2581 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2641 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2701 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2761 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2821 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2881 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2941 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3001 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3061 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3121 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3181 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3241 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3301 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3361 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     3421 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     3481 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3541 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3601 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     3661 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     3721 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     3781 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     3841 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     3901 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     3961 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     4021 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     4081 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     4141 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     4201 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     4261 gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct gctggagatg gcggacgcga
     4321 tggatatgtt ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg
     4381 ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga
     4441 ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc
     4501 gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac
     4561 gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg
     4621 tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat
     4681 gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc
     4741 cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct gcttctcgcc
     4801 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     4861 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     4921 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     4981 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     5041 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
     5101 gcgctcactg cccgctttcc agtcgggaaa ctgtcgtgcc agctgcatta atgaatcggc
     5161 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     5221 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     5281 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     5341 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     5401 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     5461 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     5521 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     5581 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     5641 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     5701 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     5761 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     5821 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     5881 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     5941 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     6001 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     6061 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     6121 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     6181 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     6241 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     6301 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     6361 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     6421 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     6481 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     6541 cgtagaggat cgagatctcg atcccgcgaa at
//
