LOCUS       sequence-208683-        9205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses pentamerised full-length protein ectodomain with no 
            N-linked glycans in mammalian cells. Contains a C-terminal rat Cd4 
            d3+4 tag, COMP pentamerisation domain, flag- and 6xHis tags. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9205)
  AUTHORS   Knockel J, Dundas K, Yang ASP, Galaway F, Metcalf T, Gemert GV, 
            Sauerwein RW, Rayner JC, Billker O, Wright GJ
  TITLE     Systematic Identification of Plasmodium Falciparum Sporozoite 
            Membrane Protein Interactions Reveals an Essential Role for the p24 
            Complex in Host Infection.
  JOURNAL   Mol Cell Proteomics. 2021;20:100038. doi: 10.1074/mcp.RA120.002432. 
            Epub 2021 Jan 27.
  PUBMED    33515807
REFERENCE   2  (bases 1 to 9205)
  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: "Mol Cell 
            Proteomics."; date: "2021"; volume: "20:100038. doi"; pages: " 
            10.1074/mcp.RA120.002432. Epub 2021 Jan 27"
COMMENT     Sequence Label: PIESP15-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     310..329
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      352..361
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2737..3522
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
                     FKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     CDS             3529..3594
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3601..3618
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3656..3711
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3710..3729)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5947..5965)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6033..6137
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6138..6206,6207..6998)
                     /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"
     rep_origin      7169..7757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7658..7677
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7911..7928
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8045..8066
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8081..8086,8087..8104,8105..8111)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8119..8135
                     /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     8124..8146
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8143..8159
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8143..8159
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8430..8809
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8810..9013
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8964..8984
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9010..9034
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 ctcgagaaag gcgtctaacc agtcacagtc gcaaggtagg ctgagcaccg tggcgggcgg
       61 cagcgggtgg cggtcggggt tgtttctggc ggaggtgctg ctgatgatgt aattaaagta
      121 ggcggtcttg agacggcgga tggtcgaggt gaggtgtggc aggcttgaga tccagctgtt
      181 ggggtgagta ctccctctca aaagcgggca ttacttctgc gctaagattg tcagtttcca
      241 aaaacgagga ggatttgata ttcacctggc ccgatctggc catacacttg agtgacaatg
      301 acatccactt tgcctttctc tccacaggtg tccactccca ggtccaagtt tgccaccatg
      361 gagtttcaga cccaggtact catgtccctg ctgctctgca tgtctggtgc ggccgccaag
      421 aacagcgaca ccaagaacaa cgcctacaac gactactaca acacctacct gagcaagatc
      481 cacagcttca aggccccttt caacgtcgag aacctggaaa acggctggaa cgtggactac
      541 agcagcgtgt ccaccaacaa gcacgtggtg ctgatcccca gcatctacaa cagacggggc
      601 gtgctgtaca acaagaaccc catcaactcc gacaacttca tcatcgagtt caatttcttc
      661 atggaacgga agtactacaa agagaagatc gacgagaagt actataagga cgagaagaac
      721 gacgtgcacg agatcgtgta caagaccaac agcgacgaca acctggccac cagcaacaag
      781 gccaacaaca aagagcagaa gcagatcaag cggaacggct tcgccctgtg gatcctgaag
      841 gaccccttcg tgatcaaaga ggccggcagc aacgtcaacc tggtgctgca ggaagaagag
      901 tacaacatct tcggctacaa gaacaggttc aacggcatcg gcatcctgtt ccagctgaag
      961 aagtctacca acaaagctaa caataaggct aataacaaca acaattacta ctactataac
     1021 aactacaacg atatcctgaa cgtggccatc atcctgaaca acggcgacaa gagcatcaat
     1081 agcgacgact tcaccatctt caagaacatc cccatcgaca tgattatgat caacaacctg
     1141 atccacacca agatcatctg caacaagaaa gagatcatca ttgccttcta ctaccccaag
     1201 agcaacaact atatcaacac cctcatcatg aagaacaata tcgccaccga gaattatatc
     1261 gccttcacct ccttcaatta caaagaggac gacaacaaaa acatcgacta caatgacgtg
     1321 aacaacaagt attacgagaa ccagaacggc tacctgccca ccttcgtggg catcaaacag
     1381 gtgctgacct tcaacctgaa caacaacaat aacaacaaca acaatatcaa catcaatgag
     1441 gccaacatta acaataataa ttataacaac aacaacaaca atgataacaa caagatcaac
     1501 tacgaccaga actacaacaa ggacatcagc gccaacgaca tcaacgccgc cttcaacgat
     1561 gtgctcaaca acatcaaccc cgagaagaat accatcaacc aggccatcga cgtgctgaac
     1621 agcatcacca agctgctgca gaaattcatg aaccaccaga tcaacagcga gaagaagctc
     1681 ctccagaaca tcaacatcct caacgagaag ctgcagttcg tgcagaccga gctgaaagtg
     1741 atcaagaaga atctggccaa tcaggccgag gaacccaaga acctgcagaa gatgttctcc
     1801 acagagctga ccggcctgaa gaacctgttt cacacccaca cccagcacca caagaagaac
     1861 atcgaggaca tcacaaacaa gctgaccaag aagattgagc ggaacaacga gctgaagatc
     1921 ctgaccgaga aggcccagaa actggaacag atcatcaaca agggcgcgcc gtcgacctcc
     1981 atcacggcct ataagagtga gggggagtca gcggagttct ccttcccact caaccttgga
     2041 gaggaaagcc tgcagggaga gttgagatgg aaggcagaga aggctccttc ttcccagtcc
     2101 tggatcacct tctccctaaa gaaccaaaag gtgtctgtgc agaagtctac tagcaacccc
     2161 aagttccagc tgtccgaaac gctcccactc acccttcaga taccccaggt ctcccttcag
     2221 tttgctggtt ctggcaacct gaccctgact ctggacagag ggatactgta tcaggaagtg
     2281 aacctggtgg tgatgaaagt gactcagccc gacagcaaca ctttgacctg tgaggtgatg
     2341 ggacccacct cacccaagat gagactgatc ttgaagcagg agaatcagga ggccagggtc
     2401 tccaggcagg agaaagtgat tcaagtgcag gcccctgaag caggggtgtg gcaatgtcta
     2461 ctgagtgaag gtgaagaggt caagatggac tccaagatcc aggttttatc caaagggttg
     2521 aattccggcg gcggctccgg aggcggtacc ggtggcggcg acctagcccc acagatgctt
     2581 cgagaactcc aggagactaa tgcggcgctg caagacgtga gagagctctt gcgacagcag
     2641 gtcaaggaga tcaccttcct gaagaatacg gtgatggaat gtgacgcctg cggtctagac
     2701 agaaacctgc ctccactagc acctctgggc ccacacccag aaacgctggt gaaagtaaaa
     2761 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2821 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2881 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2941 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3001 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3061 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3121 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3181 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3241 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3301 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3361 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     3421 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3481 agacagatcg ctgagatagg tgcctcactg attaagcatt ggggccctga ctacaaagac
     3541 catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga caagggccct
     3601 caccaccacc atcaccacta attaattaag gatcccccga cctcgacctc tggctaataa
     3661 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     3721 acatatggga gggcaaatca tttggtcgag atccctcgga gatctctagc tagaggatcg
     3781 atccccgccc cggacgaact aaacctgact acgacatctc tgccccttct tcgcggggca
     3841 gtgcatgtaa tcccttcagt tggttggtac aacttgccaa ctgggccctg ttccacatgt
     3901 gacacggggg gggaccaaac acaaaggggt tctctgactg tagttgacat ccttataaat
     3961 ggatgtgcac atttgccaac actgagtggc tttcatcctg gagcagactt tgcagtctgt
     4021 ggactgcaac acaacattgc ctttatgtgt aactcttggc tgaagctctt acaccaatgc
     4081 tgggggacat gtacctccca ggggcccagg aagactacgg gaggctacac caacgtcaat
     4141 cagaggggcc tgtgtagcta ccgataagcg gaccctcaag agggcattag caatagtgtt
     4201 tataaggccc ccttgttaac cctaaacggg tagcatatgc ttcccgggta gtagtatata
     4261 ctatccagac taaccctaat tcaatagcat atgttaccca acgggaagca tatgctatcg
     4321 aattagggtt agtaaaaggg tcctaaggaa cagcgatatc tcccacccca tgagctgtca
     4381 cggttttatt tacatggggt caggattcca cgagggtagt gaaccatttt agtcacaagg
     4441 gcagtggctg aagatcaagg agcgggcagt gaactctcct gaatcttcgc ctgcttcttc
     4501 attctccttc gtttagctaa tagaataact gctgagttgt gaacagtaag gtgtatgtga
     4561 ggtgctcgaa aacaaggttt caggtgacgc ccccagaata aaatttggac ggggggttca
     4621 gtggtggcat tgtgctatga caccaatata accctcacaa accccttggg caataaatac
     4681 tagtgtagga atgaaacatt ctgaatatct ttaacaatag aaatccatgg ggtggggaca
     4741 agccgtaaag actggatgtc catctcacac gaatttatgg ctatgggcaa cacataatcc
     4801 tagtgcaata tgatactggg gttattaaga tgtgtcccag gcagggacca agacaggtga
     4861 accatgttgt tacactctat ttgtaacaag gggaaagaga gtggacgccg acagcagcgg
     4921 actccactgg ttgtctctaa cacccccgaa aattaaacgg ggctccacgc caatggggcc
     4981 cataaacaaa gacaagtggc cactcttttt tttgaaattg tggagtgggg gcacgcgtca
     5041 gcccccacac gccgccctgc ggttttggac tgtaaaataa gggtgtaata acttggctga
     5101 ttgtaacccc gctaaccact gcggtcaaac cacttgccca caaaaccact aatggcaccc
     5161 cggggaatac ctgcataagt aggtgggcgg gccaagatag gggcgcgatt gctgcgatct
     5221 ggaggacaaa ttacacacac ttgcgcctga gcgccaagca cagggttgtt ggtcctcata
     5281 ttcacgaggt cgctgagagc acggtgggct aatgttgcca tgggtagcat atactaccca
     5341 aatatctgga tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc
     5401 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt
     5461 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     5521 tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctaata
     5581 gagattaggg tagtatatgc tatcctaatt tatatctggg tagcatatac tacccaaata
     5641 tctggatagc atatgctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     5701 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     5761 tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata
     5821 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta
     5881 tccgggtagc atatgctatc ctcatgataa gctgtcaaac atgagaattt tcttgaagac
     5941 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     6001 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     6061 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     6121 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     6181 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6241 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6301 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6361 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     6421 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6481 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6541 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6601 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6661 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6721 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6781 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6841 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6901 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6961 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     7021 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     7081 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7141 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7201 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7261 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7321 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7381 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7441 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7501 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7561 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7621 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7681 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7741 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7801 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7861 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     7921 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     7981 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     8041 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     8101 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     8161 atgattacgc caagctctag ctagaggtcg accaattctc atgtttgaca gcttatcatc
     8221 gcagatccgg gcaacgttgt tgccattgct gcaggcgcag aactggtagg tatggaagat
     8281 ctatacattg aatcaatatt ggcaattagc catattagtc attggttata tagcataaat
     8341 caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat
     8401 tggctcatgt ccaatatgac cgccatgttg acattgatta ttgactagtt attaatagta
     8461 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     8521 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     8581 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     8641 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat
     8701 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga
     8761 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     8821 ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     8881 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     8941 tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     9001 tataagcaga gctcgtttag tgaaccgtca gatcctcact ctcttccgca tcgctgtctg
     9061 cgagggccag ctgttgggct cgcggttgag gacaaactct tcgcggtctt tccagtactc
     9121 ttggatcgga aacccgtcgg cctccgaacg gtactccgcc accgagggac ctgagcgagt
     9181 ccgcatcgac cggatcggaa aacct
//
