LOCUS       sequence-203827-        7599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses enzymatically monobiotinylated full-length BMR1_04g09435 
            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 7599)
  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 7599)
  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_04g09435-bio-His
FEATURES             Location/Qualifiers
     source          1..7599
                     /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             2763..2780
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2832..2887
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2886..2905)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5123..5141)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5209..5313
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5314..5382,5383..6174)
                     /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(5532..5551)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6345..6933
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6834..6853
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7087..7104
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7221..7242
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7257..7262,7263..7280,7281..7287)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7295..7311
                     /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     7300..7322
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7319..7335
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7319..7335
                     /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 gcggccgcca
     1201 agaacgtgaa ccgggccatc cggtccaagg tgtacaacaa ccacgccaac gacatgccca
     1261 tcaaccccgt gaaggtgcag caccagaagc acaacccctg gcggaccaag ttcatgaagt
     1321 tcgacagcga caacgaggaa gagggcaacg agaacgacca cgacgaggac aatgtcgaag
     1381 agggggagga agtgctggac agcgagaagt ccaagaccag cagccacatc gtgaaccctg
     1441 ccggcaccag caaggacgac aacaccatcg agaatgacaa tgagtacaag cccagagtgt
     1501 gcggccaggg caagcggggc atcctgatct atcagcagca ggactacgga atcgccatca
     1561 tgggcctgat cgacaagggc accctgaagc tgtacgccgg caacatcatc ctgaaagaaa
     1621 tcagcctgtg gaatatcatc agccctatcg agatggccaa caacaagtgc ttcagcatcc
     1681 ggcagcccac ccagctgccc accattctgt gtgccggcgg aatcgacagc cggaacagat
     1741 gggtcaacgc cctggaaagc agccggctgt gcctgatcac caaagtgaag tactacctgc
     1801 ccatctccat cgataaggac ttcgtggaac ccgaggaccc accccccagc ggcatcaacg
     1861 tgttcattgc cgagagccag ttcggcaagc ccgagatcgt gatcaacggc aagaccctcg
     1921 agcagatcaa gaacgacgag atggaatcca tcagcacctc cgacgagtac ggcaccacac
     1981 tgtggaacgg cgagctgcag aagcccgact tccaggaaga tgagttcaac ggcgacgaga
     2041 gcatggaaag cctggccgag gccgagatga agaaggatat cgcccccgag ttcggccacg
     2101 cccggatgtt cggcgcgccg tcgacctcca tcacggccta taagagtgag ggggagtcag
     2161 cggagttctc cttcccactc aaccttggag aggaaagcct gcagggagag ttgagatgga
     2221 aggcagagaa ggctccttct tcccagtcct ggatcacctt ctccctaaag aaccaaaagg
     2281 tgtctgtgca gaagtctact agcaacccca agttccagct gtccgaaacg ctcccactca
     2341 cccttcagat accccaggtc tcccttcagt ttgctggttc tggcaacctg accctgactc
     2401 tggacagagg gatactgtat caggaagtga acctggtggt gatgaaagtg actcagcccg
     2461 acagcaacac tttgacctgt gaggtgatgg gacccacctc acccaagatg agactgatct
     2521 tgaagcagga gaatcaggag gccagggtct ccaggcagga gaaagtgatt caagtgcagg
     2581 cccctgaagc aggggtgtgg caatgtctac tgagtgaagg tgaagaggtc aagatggact
     2641 ccaagatcca ggttttatcc aaagggttga attccggcag cctgcaccac atcctggacg
     2701 cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc cctctgggcc
     2761 cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc ccccgacctc
     2821 gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     2881 tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc
     2941 tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc
     3001 ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg
     3061 gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt
     3121 tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc
     3181 agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa
     3241 gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg
     3301 ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
     3361 cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc
     3421 cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg
     3481 gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc
     3541 accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac
     3601 cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
     3661 cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac
     3721 agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat
     3781 ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc
     3841 cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat
     3901 ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
     3961 gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag
     4021 ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg
     4081 acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct
     4141 ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga
     4201 gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
     4261 gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa
     4321 accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc
     4381 gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg
     4441 gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg
     4501 tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
     4561 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     4621 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     4681 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     4741 atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc
     4801 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
     4861 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     4921 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     4981 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     5041 gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga
     5101 gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5161 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5221 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5281 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5341 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5401 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5461 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5521 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     5581 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5641 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5701 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5761 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5821 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     5881 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5941 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6001 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6061 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6121 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6181 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6241 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6301 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6361 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6421 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6481 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6541 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6601 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6661 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6721 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6781 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6841 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6901 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6961 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7021 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     7081 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     7141 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     7201 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7261 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7321 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt
     7381 ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact
     7441 ggtaggtatg gaagatctat acattgaatc aatattggca attagccata ttagtcattg
     7501 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
     7561 atatgtacat ttatattggc tcatgtccaa tatgaccgc
//
