LOCUS       sequence-76310-e        7959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses enzymatically monobiotinylated full length PFB0475c.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7959)
  AUTHORS   Zenonos ZA, Rayner JC, Wright GJ
  TITLE     Towards a comprehensive Plasmodium falciparum merozoite cell surface
            and secreted recombinant protein library.
  JOURNAL   Malar J. 2014 Mar 12;13(1):93. doi: 10.1186/1475-2875-13-93.
  PUBMED    24620899
REFERENCE   2  (bases 1 to 7959)
  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"; journalName: "Malar J."; 
            date: "2014-03-12"; volume: "13(1):93. doi"; pages: " 
            10.1186/1475-2875-13-93"
COMMENT     Sequence Label: PFB0475c-bio-His
FEATURES             Location/Qualifiers
     source          1..7959
                     /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             3123..3140
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3192..3247
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3246..3265)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5483..5501)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5569..5673
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5674..5742,5743..6534)
                     /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(5892..5911)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6705..7293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7194..7213
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7447..7464
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7581..7602
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7617..7622,7623..7640,7641..7647)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7655..7671
                     /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     7660..7682
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7679..7695
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7679..7695
                     /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 gcggccgcac
     1201 tgtgcatctg cagaaacaga gagctgttcc cccacatcag cgagaacaag cccatcggcc
     1261 agctgtaccg gctgatgaac atccacaaat acgagagctt cagcatcatc atccagatcc
     1321 accacctgaa cctgaagttc ggcgacgacg acaacgccaa gttcatcgtg cacctgaaga
     1381 tcggaaaccg ctacgcctac acccactacc acaagcagta ccagaacaag gtgcacatcg
     1441 aggaacggaa gaacatggtg gtcaagcaga acaacaacgc cctgcggctg gaagtgtaca
     1501 agaagggaac cctgaagaac accttcttcg gcagcgccga gatccacatc tacagcgaga
     1561 tcgtgaagaa gctgtttccc tgcaacgtgt acttcaatat cgccaacaag aaccagatcg
     1621 tgggcaccgc ctgcctgagc ttccactaca tcaacctgga ctgcatcaag aaggacgacc
     1681 agatctacac cagcctgttc atcgagacaa tcatcagcgt gcagaagaac caggccaaga
     1741 acaacgagaa gatcgagaag ctgatcgacg agggcaaaga gcacttcgag gccatcaaag
     1801 agacagacct gagcaccacc atctacaaga acattgccaa cctggtgctc gaggacaaga
     1861 tccggctgtt ctgcaagaac ctgaacggct acctgctgca cagcaacttc tacatcaagc
     1921 ggttctacaa caagtactac ttctacctgc acttcttcaa gggcaagttc tactggtgct
     1981 actacaacga agaggccgac gccaagatgg acaagaacag agtgggctac gtgcgcctgg
     2041 aatacgtggc caatgtgtac agcgacgtgt acagccacaa gtatttttac atcaagtacc
     2101 ggaagaagaa cgagcggaaa gagaactacc tgtacctcaa gaccatcgac aaggaccgga
     2161 acatctgggt caacatcatc cacgacttca tcatcctggt gtccaactac aagcgcgagc
     2221 gcaagaacaa gaagtacaag atcaaagagt tcaaggacaa cctgatcgag gacaccccca
     2281 aagagatcct cgagatcaac aaggccctga gcagaagcct gtctaacaac gccatgaaga
     2341 acaagtacct ggacaaaaag aagaaggtcg aggtcctgag cgacatggac aatgaagaga
     2401 attacatgaa cagcggcgac ctgggccccg tgttcaagga tatgagcaag aacatgtaca
     2461 actacgccga tggcgcgccg tcgacctcca tcacggccta taagagtgag ggggagtcag
     2521 cggagttctc cttcccactc aaccttggag aggaaagcct gcagggagag ttgagatgga
     2581 aggcagagaa ggctccttct tcccagtcct ggatcacctt ctccctaaag aaccaaaagg
     2641 tgtctgtgca gaagtctact agcaacccca agttccagct gtccgaaacg ctcccactca
     2701 cccttcagat accccaggtc tcccttcagt ttgctggttc tggcaacctg accctgactc
     2761 tggacagagg gatactgtat caggaagtga acctggtggt gatgaaagtg actcagcccg
     2821 acagcaacac tttgacctgt gaggtgatgg gacccacctc acccaagatg agactgatct
     2881 tgaagcagga gaatcaggag gccagggtct ccaggcagga gaaagtgatt caagtgcagg
     2941 cccctgaagc aggggtgtgg caatgtctac tgagtgaagg tgaagaggtc aagatggact
     3001 ccaagatcca ggttttatcc aaagggttga attccggcag cctgcaccac atcctggacg
     3061 cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc cctctgggcc
     3121 cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc ccccgacctc
     3181 gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     3241 tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc
     3301 tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc
     3361 ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg
     3421 gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt
     3481 tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc
     3541 agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa
     3601 gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg
     3661 ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
     3721 cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc
     3781 cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg
     3841 gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc
     3901 accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac
     3961 cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
     4021 cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac
     4081 agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat
     4141 ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc
     4201 cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat
     4261 ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
     4321 gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag
     4381 ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg
     4441 acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct
     4501 ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga
     4561 gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
     4621 gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa
     4681 accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc
     4741 gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg
     4801 gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg
     4861 tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
     4921 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     4981 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     5041 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     5101 atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc
     5161 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
     5221 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     5281 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     5341 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     5401 gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga
     5461 gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5521 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5581 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5641 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5701 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5761 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5821 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5881 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     5941 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6001 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6061 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6121 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6181 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     6241 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6301 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6361 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6421 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6481 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6541 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6601 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6661 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6721 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6781 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6841 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6901 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6961 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7021 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7081 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7141 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7201 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7261 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7321 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7381 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     7441 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     7501 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     7561 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7621 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7681 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt
     7741 ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact
     7801 ggtaggtatg gaagatctat acattgaatc aatattggca attagccata ttagtcattg
     7861 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
     7921 atatgtacat ttatattggc tcatgtccaa tatgaccgc
//
