LOCUS       sequence-213060-        7269 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses enzymatically monobiotinylated full-length BMR1_03g01685 
            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 7269)
  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 7269)
  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_03g01685-bio-His
FEATURES             Location/Qualifiers
     source          1..7269
                     /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             2433..2450
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2502..2557
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2556..2575)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4793..4811)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4879..4983
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4984..5052,5053..5844)
                     /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(5202..5221)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6015..6603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6504..6523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6757..6774
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6891..6912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6927..6932,6933..6950,6951..6957)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6965..6981
                     /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     6970..6992
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6989..7005
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6989..7005
                     /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 gcggcatctt cgagcggaac ttccccaaca gattcgcctg cgtgaacgac atctacaaga
     1261 tcgccagcaa gaactacgcc atcaagcccc aggacctgcc caacgtgccc ctgaacgtgg
     1321 cccgggtgtc ctacgacatg gaaaccctgc agaacaccgc catcgagaga gagctgagcg
     1381 agatcctgct gaagctgcgg gacgagtcca agaacctgga agtgggcctg acctacccct
     1441 accagtacca gcggttcctg gacgccctgg ccaaggacta cgtgtacgtg accagcgagc
     1501 tgacccagct gcatctgctg aaaaccctga catacgacag cagcgtgtac cggctgttcg
     1561 cctccaacag ctacaaccac ttcaacggca agaccaccga gattgccaag ttcaacctgt
     1621 gcagcgccat cgtgtacgcc aatgtggcca acatcatcag cgaccgggtg gacaaggtgt
     1681 acaagagcat caacaaggcc aagagccggg tgcagatcaa cctgatcaat ctgcccaccc
     1741 tgaagaagga cggcctgatc agcaagtaca tcgacagcgc cggcgcgccg tcgacctcca
     1801 tcacggccta taagagtgag ggggagtcag cggagttctc cttcccactc aaccttggag
     1861 aggaaagcct gcagggagag ttgagatgga aggcagagaa ggctccttct tcccagtcct
     1921 ggatcacctt ctccctaaag aaccaaaagg tgtctgtgca gaagtctact agcaacccca
     1981 agttccagct gtccgaaacg ctcccactca cccttcagat accccaggtc tcccttcagt
     2041 ttgctggttc tggcaacctg accctgactc tggacagagg gatactgtat caggaagtga
     2101 acctggtggt gatgaaagtg actcagcccg acagcaacac tttgacctgt gaggtgatgg
     2161 gacccacctc acccaagatg agactgatct tgaagcagga gaatcaggag gccagggtct
     2221 ccaggcagga gaaagtgatt caagtgcagg cccctgaagc aggggtgtgg caatgtctac
     2281 tgagtgaagg tgaagaggtc aagatggact ccaagatcca ggttttatcc aaagggttga
     2341 attccggcag cctgcaccac atcctggacg cccagaagat gctgtggaac cacagggaca
     2401 ggaacctgcc tcctctggcc cctctgggcc cccaccacca tcaccaccat tgagtctaga
     2461 cacgtgatta attaaggatc ccccgacctc gacctctggc taataaagga aatttatttt
     2521 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     2581 aaatcatttg gtcgagatcc ctcggagatc tctagctaga ggatcgatcc ccgccccgga
     2641 cgaactaaac ctgactacga catctctgcc ccttcttcgc ggggcagtgc atgtaatccc
     2701 ttcagttggt tggtacaact tgccaactgg gccctgttcc acatgtgaca cgggggggga
     2761 ccaaacacaa aggggttctc tgactgtagt tgacatcctt ataaatggat gtgcacattt
     2821 gccaacactg agtggctttc atcctggagc agactttgca gtctgtggac tgcaacacaa
     2881 cattgccttt atgtgtaact cttggctgaa gctcttacac caatgctggg ggacatgtac
     2941 ctcccagggg cccaggaaga ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg
     3001 tagctaccga taagcggacc ctcaagaggg cattagcaat agtgtttata aggccccctt
     3061 gttaacccta aacgggtagc atatgcttcc cgggtagtag tatatactat ccagactaac
     3121 cctaattcaa tagcatatgt tacccaacgg gaagcatatg ctatcgaatt agggttagta
     3181 aaagggtcct aaggaacagc gatatctccc accccatgag ctgtcacggt tttatttaca
     3241 tggggtcagg attccacgag ggtagtgaac cattttagtc acaagggcag tggctgaaga
     3301 tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc ttcttcattc tccttcgttt
     3361 agctaataga ataactgctg agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca
     3421 aggtttcagg tgacgccccc agaataaaat ttggacgggg ggttcagtgg tggcattgtg
     3481 ctatgacacc aatataaccc tcacaaaccc cttgggcaat aaatactagt gtaggaatga
     3541 aacattctga atatctttaa caatagaaat ccatggggtg gggacaagcc gtaaagactg
     3601 gatgtccatc tcacacgaat ttatggctat gggcaacaca taatcctagt gcaatatgat
     3661 actggggtta ttaagatgtg tcccaggcag ggaccaagac aggtgaacca tgttgttaca
     3721 ctctatttgt aacaagggga aagagagtgg acgccgacag cagcggactc cactggttgt
     3781 ctctaacacc cccgaaaatt aaacggggct ccacgccaat ggggcccata aacaaagaca
     3841 agtggccact cttttttttg aaattgtgga gtgggggcac gcgtcagccc ccacacgccg
     3901 ccctgcggtt ttggactgta aaataagggt gtaataactt ggctgattgt aaccccgcta
     3961 accactgcgg tcaaaccact tgcccacaaa accactaatg gcaccccggg gaatacctgc
     4021 ataagtaggt gggcgggcca agataggggc gcgattgctg cgatctggag gacaaattac
     4081 acacacttgc gcctgagcgc caagcacagg gttgttggtc ctcatattca cgaggtcgct
     4141 gagagcacgg tgggctaatg ttgccatggg tagcatatac tacccaaata tctggatagc
     4201 atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc
     4261 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc
     4321 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     4381 atatgctatc ctaatctgta tccgggtagc atatgctatc ctaatagaga ttagggtagt
     4441 atatgctatc ctaatttata tctgggtagc atatactacc caaatatctg gatagcatat
     4501 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagcatag
     4561 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     4621 gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     4681 gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat
     4741 gctatcctca tgataagctg tcaaacatga gaattttctt gaagacgaaa gggcctcgtg
     4801 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4861 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4921 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4981 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5041 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5101 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5161 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5221 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5281 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5341 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5401 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5461 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5521 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5581 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5641 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5701 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5761 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5821 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5881 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5941 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6001 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6061 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6721 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6781 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6841 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     6901 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     6961 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7021 ctctagctag aggtcgacca attctcatgt ttgacagctt atcatcgcag atccgggcaa
     7081 cgttgttgcc attgctgcag gcgcagaact ggtaggtatg gaagatctat acattgaatc
     7141 aatattggca attagccata ttagtcattg gttatatagc ataaatcaat attggctatt
     7201 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
     7261 tatgaccgc
//
