LOCUS       sequence-203845-        7563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses enzymatically monobiotinylated full-length BMR1_01g03475 
            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 7563)
  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 7563)
  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: 
            "10.1016/j.ijpara.2018.10.003"; journalName: "Int J Parasitol"; 
            date: "2019-02"; volume: "49"; issue: "2"; pages: "115-125"
COMMENT     Sequence Label: BMR1_01g03475-bio-His
FEATURES             Location/Qualifiers
     source          1..7563
                     /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             2727..2744
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2796..2851
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2850..2869)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5087..5105)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5173..5277
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5278..5346,5347..6138)
                     /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(5496..5515)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6309..6897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6798..6817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7051..7068
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7185..7206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7221..7226,7227..7244,7245..7251)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7259..7275
                     /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     7264..7286
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7283..7299
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7283..7299
                     /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 gcggccgccg
     1201 ctacccccaa tctgcctgtg gacagctacg ccttcgtgga caagaagaac gccaagctga
     1261 acaagaagcg gcccagcaag cccctgcgga gcagacccgt gaagaaagcc gtgtcccggc
     1321 acaagtacaa ctacaagccc aagaaaaccg tggccaacgc ccctgccaag agcaacatca
     1381 agaaccccaa cgccagaccc agccggaaca agctgagcgg caagccctac ggcaacaagc
     1441 gggtgtccaa gaaccctaac accgccagag gcaaggactg ggccaagagc ggctacagca
     1501 gcaacaaggc ccacaagctg aagaacaagg ccgagctgat caacagcgcc gtgaagggcg
     1561 gcctgaacct ggtcacccct atcggcaccc tgggcctgaa cgacctgggc gccagcaagg
     1621 acaacaacga cgacgatgac gatgactacg acgacttcat cagcggccac ggcgacaaca
     1681 gcgcccacaa gagcagcggc aagatcgccg gactggccgc caatggctgg tccagcggca
     1741 agcggagcaa cgatcacgcc gccaggggcc ccaagagcga cctggaagat gactacacca
     1801 acgacgaggg cgacgacgac accaagtccg acctcgagga cgattacaca aacgatgagg
     1861 gggatgacta tgacaccaaa agcgatcttg aggatgatta taccaatgac gaaggggatt
     1921 acgacacaaa gtctgatctc gaagatgact atacaaatga tgagggggac tacgacacta
     1981 agagtgacct cgaggacgac tacactaatg acgaaggcga cggcgatagc gtgcagggca
     2041 ccaagaagaa gcccaaggcc aacagcaagg cctctggcgc gccgtcgacc tccatcacgg
     2101 cctataagag tgagggggag tcagcggagt tctccttccc actcaacctt ggagaggaaa
     2161 gcctgcaggg agagttgaga tggaaggcag agaaggctcc ttcttcccag tcctggatca
     2221 ccttctccct aaagaaccaa aaggtgtctg tgcagaagtc tactagcaac cccaagttcc
     2281 agctgtccga aacgctccca ctcacccttc agatacccca ggtctccctt cagtttgctg
     2341 gttctggcaa cctgaccctg actctggaca gagggatact gtatcaggaa gtgaacctgg
     2401 tggtgatgaa agtgactcag cccgacagca acactttgac ctgtgaggtg atgggaccca
     2461 cctcacccaa gatgagactg atcttgaagc aggagaatca ggaggccagg gtctccaggc
     2521 aggagaaagt gattcaagtg caggcccctg aagcaggggt gtggcaatgt ctactgagtg
     2581 aaggtgaaga ggtcaagatg gactccaaga tccaggtttt atccaaaggg ttgaattccg
     2641 gcagcctgca ccacatcctg gacgcccaga agatgctgtg gaaccacagg gacaggaacc
     2701 tgcctcctct ggcccctctg ggcccccacc accatcacca ccattgagtc tagacacgtg
     2761 attaattaag gatcccccga cctcgacctc tggctaataa aggaaattta ttttcattgc
     2821 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     2881 tttggtcgag atccctcgga gatctctagc tagaggatcg atccccgccc cggacgaact
     2941 aaacctgact acgacatctc tgccccttct tcgcggggca gtgcatgtaa tcccttcagt
     3001 tggttggtac aacttgccaa ctgggccctg ttccacatgt gacacggggg gggaccaaac
     3061 acaaaggggt tctctgactg tagttgacat ccttataaat ggatgtgcac atttgccaac
     3121 actgagtggc tttcatcctg gagcagactt tgcagtctgt ggactgcaac acaacattgc
     3181 ctttatgtgt aactcttggc tgaagctctt acaccaatgc tgggggacat gtacctccca
     3241 ggggcccagg aagactacgg gaggctacac caacgtcaat cagaggggcc tgtgtagcta
     3301 ccgataagcg gaccctcaag agggcattag caatagtgtt tataaggccc ccttgttaac
     3361 cctaaacggg tagcatatgc ttcccgggta gtagtatata ctatccagac taaccctaat
     3421 tcaatagcat atgttaccca acgggaagca tatgctatcg aattagggtt agtaaaaggg
     3481 tcctaaggaa cagcgatatc tcccacccca tgagctgtca cggttttatt tacatggggt
     3541 caggattcca cgagggtagt gaaccatttt agtcacaagg gcagtggctg aagatcaagg
     3601 agcgggcagt gaactctcct gaatcttcgc ctgcttcttc attctccttc gtttagctaa
     3661 tagaataact gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa aacaaggttt
     3721 caggtgacgc ccccagaata aaatttggac ggggggttca gtggtggcat tgtgctatga
     3781 caccaatata accctcacaa accccttggg caataaatac tagtgtagga atgaaacatt
     3841 ctgaatatct ttaacaatag aaatccatgg ggtggggaca agccgtaaag actggatgtc
     3901 catctcacac gaatttatgg ctatgggcaa cacataatcc tagtgcaata tgatactggg
     3961 gttattaaga tgtgtcccag gcagggacca agacaggtga accatgttgt tacactctat
     4021 ttgtaacaag gggaaagaga gtggacgccg acagcagcgg actccactgg ttgtctctaa
     4081 cacccccgaa aattaaacgg ggctccacgc caatggggcc cataaacaaa gacaagtggc
     4141 cactcttttt tttgaaattg tggagtgggg gcacgcgtca gcccccacac gccgccctgc
     4201 ggttttggac tgtaaaataa gggtgtaata acttggctga ttgtaacccc gctaaccact
     4261 gcggtcaaac cacttgccca caaaaccact aatggcaccc cggggaatac ctgcataagt
     4321 aggtgggcgg gccaagatag gggcgcgatt gctgcgatct ggaggacaaa ttacacacac
     4381 ttgcgcctga gcgccaagca cagggttgtt ggtcctcata ttcacgaggt cgctgagagc
     4441 acggtgggct aatgttgcca tgggtagcat atactaccca aatatctgga tagcatatgc
     4501 tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     4561 tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc
     4621 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     4681 tatcctaatc tgtatccggg tagcatatgc tatcctaata gagattaggg tagtatatgc
     4741 tatcctaatt tatatctggg tagcatatac tacccaaata tctggatagc atatgctatc
     4801 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagc ataggctatc
     4861 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     4921 ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     4981 ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc
     5041 ctcatgataa gctgtcaaac atgagaattt tcttgaagac gaaagggcct cgtgatacgc
     5101 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5161 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5221 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5281 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5341 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5401 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5461 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5521 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5581 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5641 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5701 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5761 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5821 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5881 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5941 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6001 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6061 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6121 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6181 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6241 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6301 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6361 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6421 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6481 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6541 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6601 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6661 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6721 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6781 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6841 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6901 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6961 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7021 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7081 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7141 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7201 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7261 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag
     7321 ctagaggtcg accaattctc atgtttgaca gcttatcatc gcagatccgg gcaacgttgt
     7381 tgccattgct gcaggcgcag aactggtagg tatggaagat ctatacattg aatcaatatt
     7441 ggcaattagc catattagtc attggttata tagcataaat caatattggc tattggccat
     7501 tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac
     7561 cgc
//
