LOCUS       sequence-76211-e        7443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses enzymatically monobiotinylated full length PFA0135w.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7443)
  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 7443)
  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: PFA0135w-bio-His
FEATURES             Location/Qualifiers
     source          1..7443
                     /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             2607..2624
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2676..2731
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2730..2749)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4967..4985)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5053..5157
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5158..5226,5227..6018)
                     /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(5376..5395)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6189..6777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6678..6697
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6931..6948
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7065..7086
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7101..7106,7107..7124,7125..7131)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7139..7155
                     /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     7144..7166
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7163..7179
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7163..7179
                     /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 tcatccagcc cgacaagcag caggaaaagg acgagctgag caacaacttc atcagcaccg
     1261 aacagctgct ggacatcagc gagaagtgga aggccagcga gtggaacaac atgttcaacc
     1321 acatccggaa cgagtgggag gacttctacc tggtgctgca gaacgacgag cggaacattc
     1381 tgcaggaaaa acacagcaac tggaatgagt ggatccagca cctcgagaac aaatggggca
     1441 acatcaacga gaacatcaac cccgagtaca agacccatct gctgcacatc tccctgacct
     1501 ggaacgagaa gcagtgggag cactgggcct acaataccct gaagtacttt ctggaaaacg
     1561 actggaacat cttcatccag gaaatcaccg aggaaatcaa cacccacatc gaccagcagt
     1621 ggatcaagtg gctgcacgag aagaacagca tgtggctgag caatgactgg attatcgacg
     1681 agaacagctt ctgggagaag atgatcaagc tggacagctc taagtacgcc tggacccagg
     1741 acgtgaagca gtactggatt aagtggaaag agcggaccaa ccaccagaac ttcatgtgga
     1801 acaattggat ccacaacaag aaccaggtca tcaatcacat gaagaaggac aagctcgagg
     1861 actgggccaa ggataagtac gacagcttca acaagtggcg cgagatcttc ctgcaggact
     1921 ggaccgccaa ccagaaatgg aagcagctgg ccaaaggcgc gccgtcgacc tccatcacgg
     1981 cctataagag tgagggggag tcagcggagt tctccttccc actcaacctt ggagaggaaa
     2041 gcctgcaggg agagttgaga tggaaggcag agaaggctcc ttcttcccag tcctggatca
     2101 ccttctccct aaagaaccaa aaggtgtctg tgcagaagtc tactagcaac cccaagttcc
     2161 agctgtccga aacgctccca ctcacccttc agatacccca ggtctccctt cagtttgctg
     2221 gttctggcaa cctgaccctg actctggaca gagggatact gtatcaggaa gtgaacctgg
     2281 tggtgatgaa agtgactcag cccgacagca acactttgac ctgtgaggtg atgggaccca
     2341 cctcacccaa gatgagactg atcttgaagc aggagaatca ggaggccagg gtctccaggc
     2401 aggagaaagt gattcaagtg caggcccctg aagcaggggt gtggcaatgt ctactgagtg
     2461 aaggtgaaga ggtcaagatg gactccaaga tccaggtttt atccaaaggg ttgaattccg
     2521 gcagcctgca ccacatcctg gacgcccaga agatgctgtg gaaccacagg gacaggaacc
     2581 tgcctcctct ggcccctctg ggcccccacc accatcacca ccattgagtc tagacacgtg
     2641 attaattaag gatcccccga cctcgacctc tggctaataa aggaaattta ttttcattgc
     2701 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     2761 tttggtcgag atccctcgga gatctctagc tagaggatcg atccccgccc cggacgaact
     2821 aaacctgact acgacatctc tgccccttct tcgcggggca gtgcatgtaa tcccttcagt
     2881 tggttggtac aacttgccaa ctgggccctg ttccacatgt gacacggggg gggaccaaac
     2941 acaaaggggt tctctgactg tagttgacat ccttataaat ggatgtgcac atttgccaac
     3001 actgagtggc tttcatcctg gagcagactt tgcagtctgt ggactgcaac acaacattgc
     3061 ctttatgtgt aactcttggc tgaagctctt acaccaatgc tgggggacat gtacctccca
     3121 ggggcccagg aagactacgg gaggctacac caacgtcaat cagaggggcc tgtgtagcta
     3181 ccgataagcg gaccctcaag agggcattag caatagtgtt tataaggccc ccttgttaac
     3241 cctaaacggg tagcatatgc ttcccgggta gtagtatata ctatccagac taaccctaat
     3301 tcaatagcat atgttaccca acgggaagca tatgctatcg aattagggtt agtaaaaggg
     3361 tcctaaggaa cagcgatatc tcccacccca tgagctgtca cggttttatt tacatggggt
     3421 caggattcca cgagggtagt gaaccatttt agtcacaagg gcagtggctg aagatcaagg
     3481 agcgggcagt gaactctcct gaatcttcgc ctgcttcttc attctccttc gtttagctaa
     3541 tagaataact gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa aacaaggttt
     3601 caggtgacgc ccccagaata aaatttggac ggggggttca gtggtggcat tgtgctatga
     3661 caccaatata accctcacaa accccttggg caataaatac tagtgtagga atgaaacatt
     3721 ctgaatatct ttaacaatag aaatccatgg ggtggggaca agccgtaaag actggatgtc
     3781 catctcacac gaatttatgg ctatgggcaa cacataatcc tagtgcaata tgatactggg
     3841 gttattaaga tgtgtcccag gcagggacca agacaggtga accatgttgt tacactctat
     3901 ttgtaacaag gggaaagaga gtggacgccg acagcagcgg actccactgg ttgtctctaa
     3961 cacccccgaa aattaaacgg ggctccacgc caatggggcc cataaacaaa gacaagtggc
     4021 cactcttttt tttgaaattg tggagtgggg gcacgcgtca gcccccacac gccgccctgc
     4081 ggttttggac tgtaaaataa gggtgtaata acttggctga ttgtaacccc gctaaccact
     4141 gcggtcaaac cacttgccca caaaaccact aatggcaccc cggggaatac ctgcataagt
     4201 aggtgggcgg gccaagatag gggcgcgatt gctgcgatct ggaggacaaa ttacacacac
     4261 ttgcgcctga gcgccaagca cagggttgtt ggtcctcata ttcacgaggt cgctgagagc
     4321 acggtgggct aatgttgcca tgggtagcat atactaccca aatatctgga tagcatatgc
     4381 tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     4441 tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc
     4501 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     4561 tatcctaatc tgtatccggg tagcatatgc tatcctaata gagattaggg tagtatatgc
     4621 tatcctaatt tatatctggg tagcatatac tacccaaata tctggatagc atatgctatc
     4681 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagc ataggctatc
     4741 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     4801 ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     4861 ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc
     4921 ctcatgataa gctgtcaaac atgagaattt tcttgaagac gaaagggcct cgtgatacgc
     4981 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5041 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5101 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5161 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5221 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5281 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5341 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5401 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5461 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5521 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5581 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5641 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5701 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5761 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5821 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5881 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5941 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6001 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6061 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6121 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6181 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6241 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6301 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6361 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6421 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6481 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6541 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6601 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6661 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6721 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6781 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6841 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6901 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6961 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7021 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7081 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7141 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag
     7201 ctagaggtcg accaattctc atgtttgaca gcttatcatc gcagatccgg gcaacgttgt
     7261 tgccattgct gcaggcgcag aactggtagg tatggaagat ctatacattg aatcaatatt
     7321 ggcaattagc catattagtc attggttata tagcataaat caatattggc tattggccat
     7381 tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac
     7441 cgc
//
