LOCUS       sequence-213070-        8349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses enzymatically monobiotinylated full-length BMR1_04g05295 
            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 8349)
  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 8349)
  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_04g05295-bio-His
FEATURES             Location/Qualifiers
     source          1..8349
                     /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             complement(1201..1218)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             3513..3530
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3582..3637
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3636..3655)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5873..5891)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5959..6063
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6064..6132,6133..6924)
                     /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(6282..6301)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7095..7683
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7584..7603
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7837..7854
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7971..7992
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8007..8012,8013..8030,8031..8037)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8045..8061
                     /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     8050..8072
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8069..8085
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8069..8085
                     /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 gcggccgccc
     1201 acaacatcct gggcaacatc cccgacaaga aaaccttccg cgaggaacag gacgtgctga
     1261 agtccgccct ggaagatggc atggtcatcc tgttcgagag cgagttcatc agccgggcca
     1321 gacccgccag cgacaacagc gccaccaacg acaacctggc ctctgaggaa cacagcaacg
     1381 ataacaacct cgagaagatc gacgataaga agcggaacga gctgcggaag gacgccaaag
     1441 tgatgagcga agtgatcctg aagcggagag agggcgagct gagcaccatc cccaagaagg
     1501 acggcaccct gcacatcatg tggaagctga tcttctacat cctgtacagc cgcagcggca
     1561 agtggctgtt caacaacccc gccgacatga acctgaagga tgccaagccc ggccaccaca
     1621 aggtgctgaa aagcacccct gcccacatcg acaacaatat ccccctgcct gagcagcaga
     1681 tctacagcat ctccaacatt cccgaggtgc tgcccgagaa gaacggcaag acccccatcc
     1741 tgctgttcgg catggaagcc ttcgtgatct tccacctgat caaccaggcc atgctggaaa
     1801 tcgtgaactc cgccgacccc atcgaggaaa aagtgctgtt ctccgagcag tgggtgcact
     1861 tcgtgaagca ccacttcttc atctttttcc catacaacgc catcaccagc aagagatgcg
     1921 tgcggctgct gtccgacaac accctggaaa gcgagatgga actgctggac gtgtgcaaga
     1981 gcctggccga gaagtccgag aaaagcgacc tgctcgagac attcatcatg atcgaccccg
     2041 agaacaatgg cgacctcctg gaaaagtgca tcaacgagaa gcacattatc cccagctaca
     2101 tgaagctcaa ccacgagaaa ctgagccatc tgcacctgaa gcacaatctg gccatggaac
     2161 tcagcgactt catctccacc tacggcatga tcatgaccaa cgccgagcag gaaatcttca
     2221 aggacaacaa cattctgaac agcaagttct ggggcgccaa gtgcaccaag gacgaggcca
     2281 tcaaggccga aaacgagatc atcaagctgt tcagcaatat ccagaaggac ctgaacggcg
     2341 tcatcaccaa gaccttccag aagctgctgg atctgctcga aaagaatggc ggcaatatca
     2401 agagcggccc tgccaagtcc ctgattaagg ctgagaacaa cgagttccag gccgccaaga
     2461 aagagtgcga gagcaacaac gaactgaaga agaacatcga agaggccaag acccagttcc
     2521 tgaagtacgc cgccgacggc agcagcggcg gagaggaact ggtcaaagaa ctggccaaca
     2581 atgagctgtc tgagtggaag ttcaacgtgg ccgctcacat tctggacacc atgaacaagg
     2641 cctgcgaaga ggacagcgcc ttctgcctgt ccgccattaa gcactgccgg aagtctctgg
     2701 aaggcctgga atccgacaca gagcagtaca agatcatcga aaagctgcac cgggccctgt
     2761 ctgccgccct gctgaaatcc ggcaagctga atgagaagga ctacattgaa gaactgagcg
     2821 gctacatcaa gcacgaccag aatgaactgt ccaccggctt cggcgcgccg tcgacctcca
     2881 tcacggccta taagagtgag ggggagtcag cggagttctc cttcccactc aaccttggag
     2941 aggaaagcct gcagggagag ttgagatgga aggcagagaa ggctccttct tcccagtcct
     3001 ggatcacctt ctccctaaag aaccaaaagg tgtctgtgca gaagtctact agcaacccca
     3061 agttccagct gtccgaaacg ctcccactca cccttcagat accccaggtc tcccttcagt
     3121 ttgctggttc tggcaacctg accctgactc tggacagagg gatactgtat caggaagtga
     3181 acctggtggt gatgaaagtg actcagcccg acagcaacac tttgacctgt gaggtgatgg
     3241 gacccacctc acccaagatg agactgatct tgaagcagga gaatcaggag gccagggtct
     3301 ccaggcagga gaaagtgatt caagtgcagg cccctgaagc aggggtgtgg caatgtctac
     3361 tgagtgaagg tgaagaggtc aagatggact ccaagatcca ggttttatcc aaagggttga
     3421 attccggcag cctgcaccac atcctggacg cccagaagat gctgtggaac cacagggaca
     3481 ggaacctgcc tcctctggcc cctctgggcc cccaccacca tcaccaccat tgagtctaga
     3541 cacgtgatta attaaggatc ccccgacctc gacctctggc taataaagga aatttatttt
     3601 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     3661 aaatcatttg gtcgagatcc ctcggagatc tctagctaga ggatcgatcc ccgccccgga
     3721 cgaactaaac ctgactacga catctctgcc ccttcttcgc ggggcagtgc atgtaatccc
     3781 ttcagttggt tggtacaact tgccaactgg gccctgttcc acatgtgaca cgggggggga
     3841 ccaaacacaa aggggttctc tgactgtagt tgacatcctt ataaatggat gtgcacattt
     3901 gccaacactg agtggctttc atcctggagc agactttgca gtctgtggac tgcaacacaa
     3961 cattgccttt atgtgtaact cttggctgaa gctcttacac caatgctggg ggacatgtac
     4021 ctcccagggg cccaggaaga ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg
     4081 tagctaccga taagcggacc ctcaagaggg cattagcaat agtgtttata aggccccctt
     4141 gttaacccta aacgggtagc atatgcttcc cgggtagtag tatatactat ccagactaac
     4201 cctaattcaa tagcatatgt tacccaacgg gaagcatatg ctatcgaatt agggttagta
     4261 aaagggtcct aaggaacagc gatatctccc accccatgag ctgtcacggt tttatttaca
     4321 tggggtcagg attccacgag ggtagtgaac cattttagtc acaagggcag tggctgaaga
     4381 tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc ttcttcattc tccttcgttt
     4441 agctaataga ataactgctg agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca
     4501 aggtttcagg tgacgccccc agaataaaat ttggacgggg ggttcagtgg tggcattgtg
     4561 ctatgacacc aatataaccc tcacaaaccc cttgggcaat aaatactagt gtaggaatga
     4621 aacattctga atatctttaa caatagaaat ccatggggtg gggacaagcc gtaaagactg
     4681 gatgtccatc tcacacgaat ttatggctat gggcaacaca taatcctagt gcaatatgat
     4741 actggggtta ttaagatgtg tcccaggcag ggaccaagac aggtgaacca tgttgttaca
     4801 ctctatttgt aacaagggga aagagagtgg acgccgacag cagcggactc cactggttgt
     4861 ctctaacacc cccgaaaatt aaacggggct ccacgccaat ggggcccata aacaaagaca
     4921 agtggccact cttttttttg aaattgtgga gtgggggcac gcgtcagccc ccacacgccg
     4981 ccctgcggtt ttggactgta aaataagggt gtaataactt ggctgattgt aaccccgcta
     5041 accactgcgg tcaaaccact tgcccacaaa accactaatg gcaccccggg gaatacctgc
     5101 ataagtaggt gggcgggcca agataggggc gcgattgctg cgatctggag gacaaattac
     5161 acacacttgc gcctgagcgc caagcacagg gttgttggtc ctcatattca cgaggtcgct
     5221 gagagcacgg tgggctaatg ttgccatggg tagcatatac tacccaaata tctggatagc
     5281 atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc
     5341 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc
     5401 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     5461 atatgctatc ctaatctgta tccgggtagc atatgctatc ctaatagaga ttagggtagt
     5521 atatgctatc ctaatttata tctgggtagc atatactacc caaatatctg gatagcatat
     5581 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagcatag
     5641 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     5701 gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     5761 gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat
     5821 gctatcctca tgataagctg tcaaacatga gaattttctt gaagacgaaa gggcctcgtg
     5881 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5941 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6001 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6061 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6121 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6181 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6241 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6301 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6361 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6421 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6481 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6541 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6601 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6661 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6721 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6781 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6841 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6901 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6961 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7021 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7081 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7141 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7201 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7261 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7321 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7381 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7441 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7501 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7561 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7621 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7681 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7741 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7801 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7861 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7921 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7981 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     8041 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     8101 ctctagctag aggtcgacca attctcatgt ttgacagctt atcatcgcag atccgggcaa
     8161 cgttgttgcc attgctgcag gcgcagaact ggtaggtatg gaagatctat acattgaatc
     8221 aatattggca attagccata ttagtcattg gttatatagc ataaatcaat attggctatt
     8281 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
     8341 tatgaccgc
//
