LOCUS       sequence-210759-        7770 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses enzymatically monobiotinylated full-length BMR1_01g00095 
            ectodomain with no N-linked glycans upon cotransfection with BirA in
            mammalian cells. Contains a C-terminal rat Cd4 d3+4 tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7770)
  AUTHORS   Elton CM, Rodriguez M, Ben Mamoun C, Lobo CA, Wright GJ
  TITLE     A library of recombinant Babesia microti cell surface and secreted 
            proteins for diagnostics discovery and reverse vaccinology.
  JOURNAL   Int J Parasitol. 2019 Feb;49(2):115-125. doi: 
            10.1016/j.ijpara.2018.10.003. Epub 2018 Oct 24.
  PUBMED    30367868
REFERENCE   2  (bases 1 to 7770)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ijpara.2018.10.003"; journalName: "Int J Parasitol"; 
            date: "2019-02"; volume: "49"; issue: "2"; pages: "115-125"
COMMENT     Sequence Label: BMR1_01g00095-bio-His
FEATURES             Location/Qualifiers
     source          1..7770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1092..1111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2934..2951
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3003..3058
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3057..3076)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5294..5312)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5380..5484
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5485..5553,5554..6345)
                     /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(5703..5722)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6516..7104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7005..7024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7258..7275
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7392..7413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7428..7433,7434..7451,7452..7458)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7466..7482
                     /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     7471..7493
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7490..7506
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7490..7506
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
      661 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
      721 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
      781 ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc
      841 ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat gtaattaaag
      901 taggcggtct tgagacggcg gatggtcgag gtgaggtgtg gcaggcttga gatccagctg
      961 ttggggtgag tactccctct caaaagcggg cattacttct gcgctaagat tgtcagtttc
     1021 caaaaacgag gaggatttga tattcacctg gcccgatctg gccatacact tgagtgacaa
     1081 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaag tttgccacca
     1141 tggagtttca gacccaggta ctcatgtccc tgctgctctg catgtctggt gcggccgcct
     1201 tcaagagccg gatcaactac tacgacatca accgggacag cagacccctg aacgccctgg
     1261 ccagaaacgg cgtgtgcgtg cccaaggacg tgggcctgta cggcaacctg aaggccgtgt
     1321 accactgcgt gtacacccgg gacggcgaga caaagaccgg cgaggaatgg gagacacatc
     1381 tgctgaaaac aaagccctgc gtggacatcg tggatatccc cggcaagccc tacaagcgga
     1441 tcgtgttcgg ccctcccatc gacaccaacg agatgatcct gagaggcggc accccctacc
     1501 tgaccctgaa tctgtgggtg cccggcatca aggccgacga catcaaagtg atcatcgact
     1561 acagcctgga cagcgcccct gccatcacca agttcaagag cctgaccaac cccatcagct
     1621 gcgagatgct gtgcgacaag caggaccagg aatattacca gtgcgccagc gagatcgacc
     1681 tggacacccc cgagacagag aacaccgcca acagcaacga ggccgtggaa atcttcgacg
     1741 agtacgacgg cagctacaac cagatcaacg agaagtccaa caagttcaac tacaagtgcg
     1801 ggaacgaggc caaagtgaag ccccccctga tcaccatcaa ctgccccaag aacaccatcg
     1861 tgctgcagtg ggagagcacc atcttcagaa gccggaagtc cgacaagagc aaagtgaaga
     1921 acacccggat ccagcggagc taccggccct tcacccacat cgacctgatg aacatcatgt
     1981 gcaactacgc caactgcgcc ctgcagttcc acgcccccct gacccacatc cctccacccc
     2041 ccagccacag aaagacccag ctgctgaagg tgtccaagga cctgcccctg ctgcccatca
     2101 gaggccacgc cctggaacag gtgcacggct ggatctacaa ctggaagaag ttcggcaaca
     2161 agtacaccaa gaaaatcatg gacgagacat acaacaccaa tgagcccatc acccccgact
     2221 tcggcgacga gctgagcaag agagagctgt acaagttcat caagagcgtg gacgaggaca
     2281 atggcgcgcc gtcgacctcc atcacggcct ataagagtga gggggagtca gcggagttct
     2341 ccttcccact caaccttgga gaggaaagcc tgcagggaga gttgagatgg aaggcagaga
     2401 aggctccttc ttcccagtcc tggatcacct tctccctaaa gaaccaaaag gtgtctgtgc
     2461 agaagtctac tagcaacccc aagttccagc tgtccgaaac gctcccactc acccttcaga
     2521 taccccaggt ctcccttcag tttgctggtt ctggcaacct gaccctgact ctggacagag
     2581 ggatactgta tcaggaagtg aacctggtgg tgatgaaagt gactcagccc gacagcaaca
     2641 ctttgacctg tgaggtgatg ggacccacct cacccaagat gagactgatc ttgaagcagg
     2701 agaatcagga ggccagggtc tccaggcagg agaaagtgat tcaagtgcag gcccctgaag
     2761 caggggtgtg gcaatgtcta ctgagtgaag gtgaagaggt caagatggac tccaagatcc
     2821 aggttttatc caaagggttg aattccggca gcctgcacca catcctggac gcccagaaga
     2881 tgctgtggaa ccacagggac aggaacctgc ctcctctggc ccctctgggc ccccaccacc
     2941 atcaccacca ttgagtctag acacgtgatt aattaaggat cccccgacct cgacctctgg
     3001 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     3061 cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag
     3121 aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg
     3181 cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc
     3241 cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct
     3301 tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc
     3361 agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca
     3421 ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa
     3481 cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa
     3541 tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta
     3601 gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat
     3661 gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga
     3721 gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt
     3781 cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg
     3841 cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg
     3901 tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg
     3961 gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa
     4021 taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt
     4081 ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac
     4141 ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga
     4201 caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca
     4261 gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa
     4321 tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca
     4381 cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact
     4441 tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat
     4501 ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct
     4561 gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt
     4621 cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata
     4681 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag cataggctat
     4741 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4801 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4861 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     4921 cctaatagag attagggtag tatatgctat cctaatttat atctgggtag catatactac
     4981 ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata tgctatccta
     5041 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     5101 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     5161 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     5221 atctgtatcc gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct
     5281 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5341 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5401 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5461 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5521 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5581 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5641 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5701 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     5761 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5821 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5881 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5941 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6001 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     6061 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6121 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6181 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6241 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6301 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6361 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6421 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6481 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6541 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6601 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6661 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6721 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6781 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6841 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6901 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6961 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7021 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7081 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7141 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7201 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7261 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7321 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7381 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7441 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     7501 tatgaccatg attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct
     7561 tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat
     7621 ggaagatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     7681 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     7741 tttatattgg ctcatgtcca atatgaccgc
//
