LOCUS       sequence-213081-        7722 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses enzymatically monobiotinylated full-length BMR1_04g08470 
            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 7722)
  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 7722)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_04g08470-bio-His
FEATURES             Location/Qualifiers
     source          1..7722
                     /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             2886..2903
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2955..3010
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3009..3028)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5246..5264)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5332..5436
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5437..5505,5506..6297)
                     /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(5655..5674)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6468..7056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6957..6976
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7210..7227
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7344..7365
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7380..7385,7386..7403,7404..7410)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7418..7434
                     /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     7423..7445
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7442..7458
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7442..7458
                     /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 gatacgcctt caagaactac cggttccccc acatcagcga gtgcggctgc atcggcaagt
     1261 gctaccggat gttcggcatc cacgaccacg agtgcttcga catcaccgtg gaagtgatcg
     1321 aggtgtccca ggtccgatgc aagaaaaagt acatcgtgga aatcgagagc ggccggttcc
     1381 actacaagac cagccacaac caggaagccg acggcaaagt cggcttctac gagaagatca
     1441 acatccacat ccggcagtgc gacacacaca tcaccctgaa gctgtacagc aaagagctgc
     1501 tgaaggacac cctcgtgggc agcctggtca tcggcgtgga cgagctgatg cgggaccagc
     1561 tgcccagacg gcggatgtac accatggaaa tcaacgacgc cagcaccagc aagatcatgc
     1621 tgagcttcta caaagtgggc gacgtgcccg acaatatcgc ccccctgatg atgcaggccc
     1681 tgctggaatg ccccaacatg gacccccggg acgtcgagta catgagcacc atcgatcagc
     1741 tgcggttctt cgccaaagtg atcaccggcc ctctgaagcg gatgaacctg ttcggcccca
     1801 cctggaccga gtactacttc gtgcccatcg agctgagcca gaccaagtgg gagtggcggt
     1861 actgggagac aagaagcaac tacctggcca agaagaaaca tatcggcgcc tacagcttcc
     1921 tgggcgtgtc caccatcgtg cctgacgaca gcaacaaggc ccacttttac gtgaagtacc
     1981 acgacaagag cggcgtgtac gacctggtgt tcaagcgggt ggaccggaac cgggacatct
     2041 ggacccaggg cctgatcgag ttcatcaccc ggcagagaga gctgatctcc aaagagagcc
     2101 tgaacgccat ctccgacatc ggcctgaaca acgagtttat caagaacagc atcaagatcg
     2161 gccggtccaa cggcaacgac aatcaggtgc tgggcgacat cattgccgac atgtctaaag
     2221 agatgatcat cagcggcgcg ccgtcgacct ccatcacggc ctataagagt gagggggagt
     2281 cagcggagtt ctccttccca ctcaaccttg gagaggaaag cctgcaggga gagttgagat
     2341 ggaaggcaga gaaggctcct tcttcccagt cctggatcac cttctcccta aagaaccaaa
     2401 aggtgtctgt gcagaagtct actagcaacc ccaagttcca gctgtccgaa acgctcccac
     2461 tcacccttca gataccccag gtctcccttc agtttgctgg ttctggcaac ctgaccctga
     2521 ctctggacag agggatactg tatcaggaag tgaacctggt ggtgatgaaa gtgactcagc
     2581 ccgacagcaa cactttgacc tgtgaggtga tgggacccac ctcacccaag atgagactga
     2641 tcttgaagca ggagaatcag gaggccaggg tctccaggca ggagaaagtg attcaagtgc
     2701 aggcccctga agcaggggtg tggcaatgtc tactgagtga aggtgaagag gtcaagatgg
     2761 actccaagat ccaggtttta tccaaagggt tgaattccgg cagcctgcac cacatcctgg
     2821 acgcccagaa gatgctgtgg aaccacaggg acaggaacct gcctcctctg gcccctctgg
     2881 gcccccacca ccatcaccac cattgagtct agacacgtga ttaattaagg atcccccgac
     2941 ctcgacctct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     3001 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag
     3061 atctctagct agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct
     3121 gccccttctt cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac
     3181 tgggccctgt tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt
     3241 agttgacatc cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg
     3301 agcagacttt gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct
     3361 gaagctctta caccaatgct gggggacatg tacctcccag gggcccagga agactacggg
     3421 aggctacacc aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga
     3481 gggcattagc aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct
     3541 tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa
     3601 cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct
     3661 cccaccccat gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg
     3721 aaccatttta gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg
     3781 aatcttcgcc tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg
     3841 aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa
     3901 aatttggacg gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa
     3961 ccccttgggc aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga
     4021 aatccatggg gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc
     4081 tatgggcaac acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg
     4141 cagggaccaa gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag
     4201 tggacgccga cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg
     4261 gctccacgcc aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt
     4321 ggagtggggg cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag
     4381 ggtgtaataa cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac
     4441 aaaaccacta atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg
     4501 ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac
     4561 agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat
     4621 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     4681 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4741 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     4801 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     4861 agcatatgct atcctaatag agattagggt agtatatgct atcctaattt atatctgggt
     4921 agcatatact acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca
     4981 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     5041 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     5101 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     5161 tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca
     5221 tgagaatttt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5281 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5341 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5401 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5461 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5521 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5581 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5641 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     5701 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5761 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5821 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5881 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5941 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     6001 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6061 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6121 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6181 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6241 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6301 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6361 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6421 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6481 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6541 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6601 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6661 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6721 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6781 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6841 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6901 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6961 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7021 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7081 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7141 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7201 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7261 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     7321 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     7381 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     7441 acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca
     7501 tgtttgacag cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga
     7561 actggtaggt atggaagatc tatacattga atcaatattg gcaattagcc atattagtca
     7621 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     7681 ataatatgta catttatatt ggctcatgtc caatatgacc gc
//
