LOCUS       sequence-76300-e        9711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses enzymatically monobiotinylated full length PFA0445w.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9711)
  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 9711)
  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: PFA0445w-bio-His
FEATURES             Location/Qualifiers
     source          1..9711
                     /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             4875..4892
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4944..4999
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4998..5017)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7235..7253)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7321..7425
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7426..7494,7495..8286)
                     /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(7644..7663)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8457..9045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8946..8965
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9199..9216
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9333..9354
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9369..9374,9375..9392,9393..9399)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9407..9423
                     /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     9412..9434
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9431..9447
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9431..9447
                     /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 gcggccgcaa
     1201 aagcccctga gacaagcctg gaaaagatca ccgagttccg gcagcagcac agaaagaccc
     1261 tggacggcag actgtgcgcc gctgccttcc tgcacgacga ccagacctac accaactgcg
     1321 ccaccagcct gagccccgat ggcacatctg gccgcgagtg gtgctacgtg gaagtgcagc
     1381 tgctgggcaa gggcagcaga gactgggact actgccggga cagcatcaac tacgacaagc
     1441 tgcggctgca cgccaagaag gtgttcgagg aaaagagcct ggaagccgac cggctgaagg
     1501 acagactgca cgtgctgaac agccgggtgt actccatgct gcagaaatac gacagcgtgt
     1561 gcggcaagaa gcacgagctg atcaacagcc ggatcgagaa gatcaacgag tggctggaca
     1621 agagcagcga gagcatcatg aagatcgaga acaacgccaa cgacctgaac agcgccaagt
     1681 ccatcatcga ccagctgcag atcgacatca aagaagagaa caagaacgtc aagcagagcg
     1741 aggaaaactg caagaacctg cccggctacg agaacgagcc tcacagcgac ggactgaagg
     1801 tgtcctactt caacaacctg ttcttcgacg gcatccccat cgagacaaga atcgagaagg
     1861 atatcaactt cctgtacaac agcagaggcc ccctggaaaa cattgccccc cacaagtaca
     1921 gcatcagata cgagggctac atcttcgtgc cccacaacgg cgcctacacc ttcaccatcg
     1981 aaaccgactg ctacatccgg ctgctggtca acaacaaagt gatcctgacc tacggcctgg
     2041 aagaggtggt ggacatgacc gacgagaaca tgtaccacta caaccagcac gatatgtacc
     2101 agtataacga cggctacaag ctgaacaccc ccttcagctt cagctacccc cagtacagcc
     2161 acatcaagta caccctgaac aacgacgacc tgagcgagat cattaagaag aactccatcc
     2221 ctatcgaact gctgggcgga gagaagcaca agttcgtgct ggaaatcagc cacagcgccc
     2281 acctgaagta caagaacgag gaaagcagca gcttcaagct ctactggaag tccagcagaa
     2341 tccaccagca gattatccag agccactact tcttcaccga gaacatcatc ccccccaccc
     2401 ggttcagcag cctggacccc gatatcttcg agctgggcct ggtggatatc aatcagcagg
     2461 tctacaagaa caacaacaac tggatcatca ccaacgtgaa caccaaatac gtgggcctgc
     2521 atctgctgaa aacccaggaa caccccacct acaacaattt cgccctgagc atcaacaccg
     2581 ccagcaacct gtttatcgcc agccccatcg acaacgtgtt ccccctgtcc ccctacaagg
     2641 acagcctgtg gaaagtgttc gacaccggcg acatcatcga gattgaaaac aagggcaaca
     2701 gcaacaacaa caacaatatc atctacaaga agaagcagtt caagatcaag ttcatccccc
     2761 tgaagaacaa ggccgagctg aagttctata tctccaacaa catcccattc ttcatcttcg
     2821 ctcagcagcg caagatcttc cccaccatct gcaacggcga gcagcagatc ctgagcagcc
     2881 ccagcaacgc cgtgttcaaa gagtgcgaag agagcagctc cctgagcaaa gagttcaact
     2941 gtatcgccgg cctgtccagc ctgcacatgg acaagaaatt ccacgtgtgg cggagcctga
     3001 actccggcat cggcgagtac atcaaagtgt ccttcaacac ccccgtgcag atcaacaagt
     3061 tccggttcaa gccccgggac gacgtgctga catggccctc tgagatcagc ctgctgttcg
     3121 acgacacaga gatcatcatt cccatcctgc acaccagcaa catcgagcac aataccgcca
     3181 agctcgagca ccccatcatc acaaccttcg tgaagattga gatcaaggac atgttcctga
     3241 accacaacga ggctggcggc tcctttgagc tgatcggcaa cagctgcaac ctggccaacg
     3301 acgattacct gacacaccac ggcgccacaa tcgatatcaa cgactgccac aacaacgctc
     3361 tgaacaatat ccccgatgtg ctgcccctga tccacggcga caagtttctg atcacctgtg
     3421 accccaactg cctggacaac ctggctggcg aggtgtacgg cagcgacgtg tactctatcg
     3481 acagcagcat ctgcaaggcc tccatccatg ccggcgtgtg caacatccag aacaagcaca
     3541 actgcaagtt cctcatcatc atcaacgaga agcagaagaa ttacatcggc accctgcaga
     3601 acagaatcct gtccctgaac caggccaaca atgccaatct ggccttcacc ttctccccaa
     3661 tcatcaaccc caacttcgcc cagttctaca gctcctaccc caacagctac agcatcgtgt
     3721 tcaagcggaa cgacgatctg aacctgccta acaaattcct gatcgactac ggcgacgtgt
     3781 tcaacaacta cggcagcttc gcctatggct ggaacaagcc tatcaccttc ctgaatagcg
     3841 cctcccagaa gaagaagtcc tacagcaacg gcctgtacag cggcggcatc ccttttccac
     3901 ccgccaccgc tagccagcac tgcatcaccg atctggaatg ccagaccaac ttctggacct
     3961 tccagaccca cgagaatggg gcctacacca tccaggtgct cgtgggaaat ctggccagcg
     4021 acatcaagca gaacaccttt atcgaagtga acggcctgcc cctcatcaag aacattcagc
     4081 tggaaaaaaa cgagtacttc gtggccgtga agaacgtcca cgtgaccagc cggtccctga
     4141 tcttcaccag cacctgtctg gaaaccgaca acgagtgcgc caatgccaag accaccatca
     4201 tggccctgca gattctgaag attggcgcgc cgtcgacctc catcacggcc tataagagtg
     4261 agggggagtc agcggagttc tccttcccac tcaaccttgg agaggaaagc ctgcagggag
     4321 agttgagatg gaaggcagag aaggctcctt cttcccagtc ctggatcacc ttctccctaa
     4381 agaaccaaaa ggtgtctgtg cagaagtcta ctagcaaccc caagttccag ctgtccgaaa
     4441 cgctcccact cacccttcag ataccccagg tctcccttca gtttgctggt tctggcaacc
     4501 tgaccctgac tctggacaga gggatactgt atcaggaagt gaacctggtg gtgatgaaag
     4561 tgactcagcc cgacagcaac actttgacct gtgaggtgat gggacccacc tcacccaaga
     4621 tgagactgat cttgaagcag gagaatcagg aggccagggt ctccaggcag gagaaagtga
     4681 ttcaagtgca ggcccctgaa gcaggggtgt ggcaatgtct actgagtgaa ggtgaagagg
     4741 tcaagatgga ctccaagatc caggttttat ccaaagggtt gaattccggc agcctgcacc
     4801 acatcctgga cgcccagaag atgctgtgga accacaggga caggaacctg cctcctctgg
     4861 cccctctggg cccccaccac catcaccacc attgagtcta gacacgtgat taattaagga
     4921 tcccccgacc tcgacctctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     4981 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt tggtcgagat
     5041 ccctcggaga tctctagcta gaggatcgat ccccgccccg gacgaactaa acctgactac
     5101 gacatctctg ccccttcttc gcggggcagt gcatgtaatc ccttcagttg gttggtacaa
     5161 cttgccaact gggccctgtt ccacatgtga cacggggggg gaccaaacac aaaggggttc
     5221 tctgactgta gttgacatcc ttataaatgg atgtgcacat ttgccaacac tgagtggctt
     5281 tcatcctgga gcagactttg cagtctgtgg actgcaacac aacattgcct ttatgtgtaa
     5341 ctcttggctg aagctcttac accaatgctg ggggacatgt acctcccagg ggcccaggaa
     5401 gactacggga ggctacacca acgtcaatca gaggggcctg tgtagctacc gataagcgga
     5461 ccctcaagag ggcattagca atagtgttta taaggccccc ttgttaaccc taaacgggta
     5521 gcatatgctt cccgggtagt agtatatact atccagacta accctaattc aatagcatat
     5581 gttacccaac gggaagcata tgctatcgaa ttagggttag taaaagggtc ctaaggaaca
     5641 gcgatatctc ccaccccatg agctgtcacg gttttattta catggggtca ggattccacg
     5701 agggtagtga accattttag tcacaagggc agtggctgaa gatcaaggag cgggcagtga
     5761 actctcctga atcttcgcct gcttcttcat tctccttcgt ttagctaata gaataactgc
     5821 tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa caaggtttca ggtgacgccc
     5881 ccagaataaa atttggacgg ggggttcagt ggtggcattg tgctatgaca ccaatataac
     5941 cctcacaaac cccttgggca ataaatacta gtgtaggaat gaaacattct gaatatcttt
     6001 aacaatagaa atccatgggg tggggacaag ccgtaaagac tggatgtcca tctcacacga
     6061 atttatggct atgggcaaca cataatccta gtgcaatatg atactggggt tattaagatg
     6121 tgtcccaggc agggaccaag acaggtgaac catgttgtta cactctattt gtaacaaggg
     6181 gaaagagagt ggacgccgac agcagcggac tccactggtt gtctctaaca cccccgaaaa
     6241 ttaaacgggg ctccacgcca atggggccca taaacaaaga caagtggcca ctcttttttt
     6301 tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc cgccctgcgg ttttggactg
     6361 taaaataagg gtgtaataac ttggctgatt gtaaccccgc taaccactgc ggtcaaacca
     6421 cttgcccaca aaaccactaa tggcaccccg gggaatacct gcataagtag gtgggcgggc
     6481 caagataggg gcgcgattgc tgcgatctgg aggacaaatt acacacactt gcgcctgagc
     6541 gccaagcaca gggttgttgg tcctcatatt cacgaggtcg ctgagagcac ggtgggctaa
     6601 tgttgccatg ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta
     6661 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     6721 tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta
     6781 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
     6841 tatccgggta gcatatgcta tcctaataga gattagggta gtatatgcta tcctaattta
     6901 tatctgggta gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc
     6961 tgggtagcat atgctatcct aatctatatc tgggtagcat aggctatcct aatctatatc
     7021 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatttatatc
     7081 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     7141 tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct catgataagc
     7201 tgtcaaacat gagaattttc ttgaagacga aagggcctcg tgatacgcct atttttatag
     7261 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     7321 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     7381 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     7441 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     7501 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     7561 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     7621 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     7681 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     7741 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     7801 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     7861 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     7921 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     7981 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     8041 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     8101 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     8161 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     8221 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     8281 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     8341 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     8401 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     8461 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     8521 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     8581 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     8641 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     8701 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     8761 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     8821 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     8881 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     8941 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     9001 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     9061 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     9121 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     9181 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     9241 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     9301 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     9361 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     9421 caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac
     9481 caattctcat gtttgacagc ttatcatcgc agatccgggc aacgttgttg ccattgctgc
     9541 aggcgcagaa ctggtaggta tggaagatct atacattgaa tcaatattgg caattagcca
     9601 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
     9661 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg c
//
