LOCUS       sequence-203824-        9462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses enzymatically monobiotinylated full-length BmR1_04g07470 
            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 9462)
  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 9462)
  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_04g07470-bio-His
FEATURES             Location/Qualifiers
     source          1..9462
                     /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             4626..4643
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4695..4750
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4749..4768)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6986..7004)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7072..7176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7177..7245,7246..8037)
                     /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(7395..7414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8208..8796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8697..8716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8950..8967
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9084..9105
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9120..9125,9126..9143,9144..9150)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9158..9174
                     /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     9163..9185
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9182..9198
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9182..9198
                     /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 tgaacgccgc ctacaacgag gccaacaccg agcagctgaa aaccggctac accttcgccg
     1261 agctgggcat gtactacgag cccaaatacg tgttcaccag ctttttcgag ggcgccagcc
     1321 ccatccacag cgccaccagc agcaactacg tgatgcacga catcgtgatc ccccggaagg
     1381 acaccaagct gaagtacttc gtgtccggca acgtgaagaa cctggtgcag ggcaagccca
     1441 tcaccgtgat gatcggcacc atgttcggca actacttcga catgctgctg gacatcaccc
     1501 tgagcatcga caacaaccgg ttcaagttcg acctgcccag aggccggtac tacatcaagc
     1561 tgcagggctc cggctatatg ctgcctggcg ccctgcggat caacctgccc tgcagaaacg
     1621 ccaagtgccc cttcgtgggc atcaactacg ccgacagcct ggaagtgtac cccaccaaga
     1681 acgaccccga cctgtacacc tttagctggc ggctgcagaa caccgcccag tacggcatcg
     1741 agacagtgtc caccgtgccc aaggacgaga gcagcctgag caataccgcc accatcaaca
     1801 attacgagaa gatcgaggcc agcgacgccg ctgccaagct ggaacacatg tttggaatcg
     1861 agctgcacgg cgtgtggggc agcgagtaca gctaccggct gctgagcgtg ctgatggaat
     1921 tccagttcct gagaagcaac aaggacaagc cccagcactg gtccctggtg gacaacgatc
     1981 tgtaccccgg cgacgtgcgg gtggtggaag aggaaaacag cagatacgag aaaatgatca
     2041 gcatcagcaa gaacgccttt aagtacgcca tcaagaaaac ccacgagcgg aagcacaacg
     2101 ggaattactt cagcagacgg ctgttcaagg ccgtgatccg ggccacctgt ctggacagcg
     2161 tcgtgaagat gaagtacctg ttcagcaaac tgcacggggt ggaaatcctg gaacccagcg
     2221 agctggacac catggcccac atgggaaagc ctatcacaaa ttacgcctgg cgggactacc
     2281 agagctggtt caagcacccc gaggaactga ttgagctggc cacctcttgg agcgagtatc
     2341 ctgccggctt ccaccggatc aagggcctga agtatctgct gcggcggatg gacggcctcg
     2401 tgaatcctga gcagcccacc gcccctgcca ttgcctaccc tagaggcccc gacaccgact
     2461 cctacatcga gttcatggaa agcggcttct acagataccc cgacatctct aagctggtgc
     2521 tgcacgaact gggccacttc atcgacatga acaccctgac cctggaactg cagatggact
     2581 ggcagcggat cggcggctgg tacaatgacg ccaacgaccc tgacggctgg tccacccggc
     2641 agcagaccca gtttgtgtcc agctacgccc acaagaagaa tccccaggaa gatttcgcca
     2701 gctccctggc cgactacctg ctgaacccca agaaactgat gaccagagcc cccgataagt
     2761 acaactggat caaaaagtcc gtgatgggcg gagcccacta tgtgaccaag tccagccacg
     2821 agttcaccgt gctgaatctg ggctccccca acttcatcta ccctggcaga atcggcagca
     2881 tcgtcatcac agtgacaggc ggcgagcggg acaacaagtt cgtgaactgg aacttcaagc
     2941 tgatctccag acagcagagc tgcgccacaa gcctgtcctt cagactgttc tccaccatcg
     3001 gcacattcgt ggacatctac ctgcggccca agggcgctaa gtgcggcaac acctttgagg
     3061 gcacccaggt gttcaactcc gccgtgaagc ggggagtgtg gaccaccgac cagatcgaga
     3121 tcaccgatca gaacggcctg cagagattcg tgggagccgc cgacttcggc ctgcgcgtgt
     3181 ggatcaacaa cgtgcacgag gacttccagg accccgtgac catcaaagag agcatcaagt
     3241 tcaatctggt gcacgacgtg gccggaccca tcctgagagt gaactacctc gtgcgggacg
     3301 acggcatcct gaaaaaccca ggcgcctcct acgccacaat caatggcgac ggctatgccc
     3361 agcacagcct gttcggctac agcgacatcc ccacccagtc ccccctgtgg accggctcca
     3421 agcctagcac ccagtgtaac acagtgcagg aggacgccta cctgaactgc tacgccgtgt
     3481 ccgtgaatgt ggtgctgagc aagaatgcca gaagcggcaa gtactctctg gccggcctga
     3541 ccaccatgga caccgccggc aacagcgaga tgctgcagtg gggcaacaat gagggcccct
     3601 acatcatgta caccagctcc ggcgtggccg acaccaacca gcccgagctg aaggacatta
     3661 ccgtgacctc caaccccgcc aacatcgata agatcgacgg cgagacaaag gtcgagatct
     3721 ccttcaagat ccgggaccag gaatccggca tctctaccgt gtggttccgg atcagggacc
     3781 ccttcggcac cgagatcctg ctgtaccctc ccgagttcga cacctccagc aacgactgga
     3841 tgacaatcaa caagatccac ctcctgcctc ggggctccgt gccaggcgtc tggcacctga
     3901 acagcatcta cgtcgaggac aacgccggaa atgccgccag agccacactg accgagaaca
     3961 tctatgtgtc tggcggcgcg ccgtcgacct ccatcacggc ctataagagt gagggggagt
     4021 cagcggagtt ctccttccca ctcaaccttg gagaggaaag cctgcaggga gagttgagat
     4081 ggaaggcaga gaaggctcct tcttcccagt cctggatcac cttctcccta aagaaccaaa
     4141 aggtgtctgt gcagaagtct actagcaacc ccaagttcca gctgtccgaa acgctcccac
     4201 tcacccttca gataccccag gtctcccttc agtttgctgg ttctggcaac ctgaccctga
     4261 ctctggacag agggatactg tatcaggaag tgaacctggt ggtgatgaaa gtgactcagc
     4321 ccgacagcaa cactttgacc tgtgaggtga tgggacccac ctcacccaag atgagactga
     4381 tcttgaagca ggagaatcag gaggccaggg tctccaggca ggagaaagtg attcaagtgc
     4441 aggcccctga agcaggggtg tggcaatgtc tactgagtga aggtgaagag gtcaagatgg
     4501 actccaagat ccaggtttta tccaaagggt tgaattccgg cagcctgcac cacatcctgg
     4561 acgcccagaa gatgctgtgg aaccacaggg acaggaacct gcctcctctg gcccctctgg
     4621 gcccccacca ccatcaccac cattgagtct agacacgtga ttaattaagg atcccccgac
     4681 ctcgacctct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     4741 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag
     4801 atctctagct agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct
     4861 gccccttctt cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac
     4921 tgggccctgt tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt
     4981 agttgacatc cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg
     5041 agcagacttt gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct
     5101 gaagctctta caccaatgct gggggacatg tacctcccag gggcccagga agactacggg
     5161 aggctacacc aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga
     5221 gggcattagc aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct
     5281 tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa
     5341 cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct
     5401 cccaccccat gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg
     5461 aaccatttta gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg
     5521 aatcttcgcc tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg
     5581 aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa
     5641 aatttggacg gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa
     5701 ccccttgggc aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga
     5761 aatccatggg gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc
     5821 tatgggcaac acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg
     5881 cagggaccaa gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag
     5941 tggacgccga cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg
     6001 gctccacgcc aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt
     6061 ggagtggggg cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag
     6121 ggtgtaataa cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac
     6181 aaaaccacta atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg
     6241 ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac
     6301 agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat
     6361 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     6421 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     6481 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     6541 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     6601 agcatatgct atcctaatag agattagggt agtatatgct atcctaattt atatctgggt
     6661 agcatatact acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca
     6721 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     6781 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     6841 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     6901 tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca
     6961 tgagaatttt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     7021 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     7081 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     7141 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     7201 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     7261 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     7321 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     7381 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     7441 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     7501 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     7561 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7621 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7681 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     7741 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7801 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7861 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7921 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7981 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     8041 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     8101 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     8161 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     8221 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     8281 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     8341 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     8401 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     8461 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     8521 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     8581 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     8641 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     8701 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     8761 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     8821 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     8881 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     8941 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     9001 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     9061 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     9121 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     9181 acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca
     9241 tgtttgacag cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga
     9301 actggtaggt atggaagatc tatacattga atcaatattg gcaattagcc atattagtca
     9361 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     9421 ataatatgta catttatatt ggctcatgtc caatatgacc gc
//
