LOCUS       sequence-210758-        8148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses enzymatically monobiotinylated full-length BMR1_03g00020 
            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 8148)
  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 8148)
  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_03g00020-bio-His
FEATURES             Location/Qualifiers
     source          1..8148
                     /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             3312..3329
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3381..3436
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3435..3454)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5672..5690)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5758..5862
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5863..5931,5932..6723)
                     /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(6081..6100)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6894..7482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7383..7402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7636..7653
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7770..7791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7806..7811,7812..7829,7830..7836)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7844..7860
                     /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     7849..7871
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7868..7884
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7868..7884
                     /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 tggataccaa caacgccgcc ctgcgcgaga gccagcccat gcctctgacc acaatcgccc
     1261 tgaccatcag catcgtgctg gacatccacg gcagccccag caacggcctg tacgagagca
     1321 acctgttcta caccgagggc tacggcaagt acctgaccag ccccaccaag atcaagacca
     1381 tcgagttcga gggatacaag ttcgagtttg acgacgacac cctgcccaag acccccatca
     1441 caaagatcca catcatcacc tacgacaaca agcccatcct gtttgagttc atgaccaaca
     1501 tgtaccggcc ctaccggcgg ttctactact acaccctgga cagcaagacc aacaagctgt
     1561 acaactacgt gaccgccgag acaggctaca acgtggaaga tagcagcggc ctgaagtact
     1621 acacagagct gagcaagagc ggcatcaacg acgtgctgca ggacctggcc aagaacatcg
     1681 acgagtccaa catcgagcac ctgaaaacca gcgacgtgat caagggcctg aatatcgcca
     1741 tcgaggtgta cagcaaccgg gtggtggaac agatcaagtc tatcaaggtg gtcacccccg
     1801 tggaactgtt cgactacaag accgaggtgt ccatcgagag cgtggaccac gagagccggg
     1861 acaacagcct ggccgaggtg gaagaggacg gcaaggccgt gcaggtcgga acccagcccg
     1921 tgtacgaagt gaacggcgcc cacaaccctg ccgcccaggt gctgagccag aacaacatca
     1981 tcgagacact ggacgacaag agcaaagtga cccacctgag aaacattgcc cacgaagtca
     2041 atatcgagct ggaacggcac gacatcagca gcctgatcaa ccaggcctcc gtggacatcg
     2101 atgacaccat caccaacaaa tactccaacg tcatcacccc tgagttcttc aagtgctacg
     2161 agtacgactc caagctcagc accaagctgt tccggtacaa gatctacaag agccagggcg
     2221 acaagctgag ccggcccaac aagatcgaga agattaagtt cttcgattac atcattgagt
     2281 tcgacgataa ggccaagctg cccatcggcg gcacagtgat caacatctac atctatacct
     2341 gcaagcacca taaccccctg ctgatccagt tcgtggtgtt caccgacgag agctgctact
     2401 attactactt ctacttcctg aatagcaaca tcaacaagtg gaacaactac aaaatcaagt
     2461 acaaccggcg gctgaaagag tacatcgaca aggacggaat cttttactac gagttcagct
     2521 gctttaagtg gtgcgtgtac ttcagcttcc tcaacaaatt cgaggacacc gtgctggccc
     2581 ggatccactg caacaagggc aaatgtgtga atgtgaacgt ggacaacatt gagaacaaga
     2641 acctcgagat ctacctgaag ggcgcgccgt cgacctccat cacggcctat aagagtgagg
     2701 gggagtcagc ggagttctcc ttcccactca accttggaga ggaaagcctg cagggagagt
     2761 tgagatggaa ggcagagaag gctccttctt cccagtcctg gatcaccttc tccctaaaga
     2821 accaaaaggt gtctgtgcag aagtctacta gcaaccccaa gttccagctg tccgaaacgc
     2881 tcccactcac ccttcagata ccccaggtct cccttcagtt tgctggttct ggcaacctga
     2941 ccctgactct ggacagaggg atactgtatc aggaagtgaa cctggtggtg atgaaagtga
     3001 ctcagcccga cagcaacact ttgacctgtg aggtgatggg acccacctca cccaagatga
     3061 gactgatctt gaagcaggag aatcaggagg ccagggtctc caggcaggag aaagtgattc
     3121 aagtgcaggc ccctgaagca ggggtgtggc aatgtctact gagtgaaggt gaagaggtca
     3181 agatggactc caagatccag gttttatcca aagggttgaa ttccggcagc ctgcaccaca
     3241 tcctggacgc ccagaagatg ctgtggaacc acagggacag gaacctgcct cctctggccc
     3301 ctctgggccc ccaccaccat caccaccatt gagtctagac acgtgattaa ttaaggatcc
     3361 cccgacctcg acctctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     3421 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttgg tcgagatccc
     3481 tcggagatct ctagctagag gatcgatccc cgccccggac gaactaaacc tgactacgac
     3541 atctctgccc cttcttcgcg gggcagtgca tgtaatccct tcagttggtt ggtacaactt
     3601 gccaactggg ccctgttcca catgtgacac ggggggggac caaacacaaa ggggttctct
     3661 gactgtagtt gacatcctta taaatggatg tgcacatttg ccaacactga gtggctttca
     3721 tcctggagca gactttgcag tctgtggact gcaacacaac attgccttta tgtgtaactc
     3781 ttggctgaag ctcttacacc aatgctgggg gacatgtacc tcccaggggc ccaggaagac
     3841 tacgggaggc tacaccaacg tcaatcagag gggcctgtgt agctaccgat aagcggaccc
     3901 tcaagagggc attagcaata gtgtttataa ggcccccttg ttaaccctaa acgggtagca
     3961 tatgcttccc gggtagtagt atatactatc cagactaacc ctaattcaat agcatatgtt
     4021 acccaacggg aagcatatgc tatcgaatta gggttagtaa aagggtccta aggaacagcg
     4081 atatctccca ccccatgagc tgtcacggtt ttatttacat ggggtcagga ttccacgagg
     4141 gtagtgaacc attttagtca caagggcagt ggctgaagat caaggagcgg gcagtgaact
     4201 ctcctgaatc ttcgcctgct tcttcattct ccttcgttta gctaatagaa taactgctga
     4261 gttgtgaaca gtaaggtgta tgtgaggtgc tcgaaaacaa ggtttcaggt gacgccccca
     4321 gaataaaatt tggacggggg gttcagtggt ggcattgtgc tatgacacca atataaccct
     4381 cacaaacccc ttgggcaata aatactagtg taggaatgaa acattctgaa tatctttaac
     4441 aatagaaatc catggggtgg ggacaagccg taaagactgg atgtccatct cacacgaatt
     4501 tatggctatg ggcaacacat aatcctagtg caatatgata ctggggttat taagatgtgt
     4561 cccaggcagg gaccaagaca ggtgaaccat gttgttacac tctatttgta acaaggggaa
     4621 agagagtgga cgccgacagc agcggactcc actggttgtc tctaacaccc ccgaaaatta
     4681 aacggggctc cacgccaatg gggcccataa acaaagacaa gtggccactc ttttttttga
     4741 aattgtggag tgggggcacg cgtcagcccc cacacgccgc cctgcggttt tggactgtaa
     4801 aataagggtg taataacttg gctgattgta accccgctaa ccactgcggt caaaccactt
     4861 gcccacaaaa ccactaatgg caccccgggg aatacctgca taagtaggtg ggcgggccaa
     4921 gataggggcg cgattgctgc gatctggagg acaaattaca cacacttgcg cctgagcgcc
     4981 aagcacaggg ttgttggtcc tcatattcac gaggtcgctg agagcacggt gggctaatgt
     5041 tgccatgggt agcatatact acccaaatat ctggatagca tatgctatcc taatctatat
     5101 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     5161 ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat
     5221 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat
     5281 ccgggtagca tatgctatcc taatagagat tagggtagta tatgctatcc taatttatat
     5341 ctgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg
     5401 gtagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg
     5461 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg
     5521 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     5581 gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctcat gataagctgt
     5641 caaacatgag aattttcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5701 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5761 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5821 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5881 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5941 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6001 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6061 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     6121 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6181 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6241 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6301 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6361 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     6421 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6481 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6541 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6601 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6661 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6721 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6781 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6841 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6901 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6961 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7021 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7081 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7141 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7201 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7261 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7321 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7381 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7441 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7501 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7561 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     7621 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     7681 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     7741 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     7801 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     7861 tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgaccaa
     7921 ttctcatgtt tgacagctta tcatcgcaga tccgggcaac gttgttgcca ttgctgcagg
     7981 cgcagaactg gtaggtatgg aagatctata cattgaatca atattggcaa ttagccatat
     8041 tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc
     8101 tatatcataa tatgtacatt tatattggct catgtccaat atgaccgc
//
