LOCUS       sequence-208681-        8665 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 8665)
  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 8665)
  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: GAP50-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8665
                     /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             2197..2982
                     /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             2989..3054
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3061..3078
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3116..3171
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3170..3189)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5407..5425)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5493..5597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5598..5666,5667..6458)
                     /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      6629..7217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7118..7137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7371..7388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7505..7526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7541..7546,7547..7564,7565..7571)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7579..7595
                     /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     7584..7606
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7603..7619
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7603..7619
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7890..8269
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8270..8473
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8424..8444
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8470..8494
                     /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 ggccgcccag
      421 ctgagatttg cctctctggg cgactggggc aaggacacca agggccagat cctgaacgcc
      481 aagtacttca agcagttcat caagaacgag cgcgtgacct tcatcgtgtc tcccggcagc
      541 aacttcatcg acggcgtgaa gggcctgaac gaccccgcct ggaagaacct gtacgaggac
      601 gtgtacagcg aagagaaggg cgatatgtac atgcctttct tcaccgtgct gggcacccgg
      661 gactggaccg gcaactacaa tgcccagctg ctgaagggcc agggcatcta catcgagaag
      721 aacggcgaga catctatcga gaaggacgcc gacgccacca actaccccaa gtggatcatg
      781 cccaactact ggtatcacta cttcacccac tttaccgtgt ccagcggccc cagcatcgtg
      841 aaaaccggcc acaaggacct ggccgctgcc ttcatcttca tcgatacctg ggtgctgagc
      901 agcaatttcc cctacaagaa gatccacgag aaggcctgga acgatctgaa gtcccagctg
      961 tccgtggcca agaagattgc cgacttcatc atcgtcgtgg gcgaccagcc catctacagc
     1021 agcggctaca gccggggcag cagctacctg gcctactacc tgctgcctct gctgaaagac
     1081 gccgaggtgg acctgtacat cagcggccac gacaacaaca tggaagtgat cgaggacaac
     1141 gacatggccc acatcacctg tggcagcggc agcatgagcc agggcaagag cggcatgaag
     1201 aacagcaaga gcctgttctt tagcagcgat atcggcttct gcgtgcacga gctgagcaac
     1261 aacggcatcg tgaccaagtt cgtgtcctcc aagaaaggcg aagtgatcta cacccacaag
     1321 ctgaacatca agaagaagaa aaccctggac aaagtgaacg ccctgcagca cttcgccgcc
     1381 ctgcccaacg tggaactgac cgacgtgcca agctccggcc ccggcgcgcc gtcgacctcc
     1441 atcacggcct ataagagtga gggggagtca gcggagttct ccttcccact caaccttgga
     1501 gaggaaagcc tgcagggaga gttgagatgg aaggcagaga aggctccttc ttcccagtcc
     1561 tggatcacct tctccctaaa gaaccaaaag gtgtctgtgc agaagtctac tagcaacccc
     1621 aagttccagc tgtccgaaac gctcccactc acccttcaga taccccaggt ctcccttcag
     1681 tttgctggtt ctggcaacct gaccctgact ctggacagag ggatactgta tcaggaagtg
     1741 aacctggtgg tgatgaaagt gactcagccc gacagcaaca ctttgacctg tgaggtgatg
     1801 ggacccacct cacccaagat gagactgatc ttgaagcagg agaatcagga ggccagggtc
     1861 tccaggcagg agaaagtgat tcaagtgcag gcccctgaag caggggtgtg gcaatgtcta
     1921 ctgagtgaag gtgaagaggt caagatggac tccaagatcc aggttttatc caaagggttg
     1981 aattccggcg gcggctccgg aggcggtacc ggtggcggcg acctagcccc acagatgctt
     2041 cgagaactcc aggagactaa tgcggcgctg caagacgtga gagagctctt gcgacagcag
     2101 gtcaaggaga tcaccttcct gaagaatacg gtgatggaat gtgacgcctg cggtctagac
     2161 agaaacctgc ctccactagc acctctgggc ccacacccag aaacgctggt gaaagtaaaa
     2221 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2281 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2341 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2401 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2461 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2521 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2581 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2641 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2701 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2761 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2821 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2881 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2941 agacagatcg ctgagatagg tgcctcactg attaagcatt ggggccctga ctacaaagac
     3001 catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga caagggccct
     3061 caccaccacc atcaccacta attaattaag gatcccccga cctcgacctc tggctaataa
     3121 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     3181 acatatggga gggcaaatca tttggtcgag atccctcgga gatctctagc tagaggatcg
     3241 atccccgccc cggacgaact aaacctgact acgacatctc tgccccttct tcgcggggca
     3301 gtgcatgtaa tcccttcagt tggttggtac aacttgccaa ctgggccctg ttccacatgt
     3361 gacacggggg gggaccaaac acaaaggggt tctctgactg tagttgacat ccttataaat
     3421 ggatgtgcac atttgccaac actgagtggc tttcatcctg gagcagactt tgcagtctgt
     3481 ggactgcaac acaacattgc ctttatgtgt aactcttggc tgaagctctt acaccaatgc
     3541 tgggggacat gtacctccca ggggcccagg aagactacgg gaggctacac caacgtcaat
     3601 cagaggggcc tgtgtagcta ccgataagcg gaccctcaag agggcattag caatagtgtt
     3661 tataaggccc ccttgttaac cctaaacggg tagcatatgc ttcccgggta gtagtatata
     3721 ctatccagac taaccctaat tcaatagcat atgttaccca acgggaagca tatgctatcg
     3781 aattagggtt agtaaaaggg tcctaaggaa cagcgatatc tcccacccca tgagctgtca
     3841 cggttttatt tacatggggt caggattcca cgagggtagt gaaccatttt agtcacaagg
     3901 gcagtggctg aagatcaagg agcgggcagt gaactctcct gaatcttcgc ctgcttcttc
     3961 attctccttc gtttagctaa tagaataact gctgagttgt gaacagtaag gtgtatgtga
     4021 ggtgctcgaa aacaaggttt caggtgacgc ccccagaata aaatttggac ggggggttca
     4081 gtggtggcat tgtgctatga caccaatata accctcacaa accccttggg caataaatac
     4141 tagtgtagga atgaaacatt ctgaatatct ttaacaatag aaatccatgg ggtggggaca
     4201 agccgtaaag actggatgtc catctcacac gaatttatgg ctatgggcaa cacataatcc
     4261 tagtgcaata tgatactggg gttattaaga tgtgtcccag gcagggacca agacaggtga
     4321 accatgttgt tacactctat ttgtaacaag gggaaagaga gtggacgccg acagcagcgg
     4381 actccactgg ttgtctctaa cacccccgaa aattaaacgg ggctccacgc caatggggcc
     4441 cataaacaaa gacaagtggc cactcttttt tttgaaattg tggagtgggg gcacgcgtca
     4501 gcccccacac gccgccctgc ggttttggac tgtaaaataa gggtgtaata acttggctga
     4561 ttgtaacccc gctaaccact gcggtcaaac cacttgccca caaaaccact aatggcaccc
     4621 cggggaatac ctgcataagt aggtgggcgg gccaagatag gggcgcgatt gctgcgatct
     4681 ggaggacaaa ttacacacac ttgcgcctga gcgccaagca cagggttgtt ggtcctcata
     4741 ttcacgaggt cgctgagagc acggtgggct aatgttgcca tgggtagcat atactaccca
     4801 aatatctgga tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc
     4861 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt
     4921 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     4981 tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctaata
     5041 gagattaggg tagtatatgc tatcctaatt tatatctggg tagcatatac tacccaaata
     5101 tctggatagc atatgctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     5161 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     5221 tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata
     5281 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta
     5341 tccgggtagc atatgctatc ctcatgataa gctgtcaaac atgagaattt tcttgaagac
     5401 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5461 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5521 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5581 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5641 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5701 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5761 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5821 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     5881 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5941 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6001 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6061 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6121 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6181 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6241 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6301 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6361 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6421 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6481 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6541 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6601 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6661 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6721 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6781 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6841 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6901 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6961 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7021 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7081 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7141 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7201 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7261 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7321 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     7381 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     7441 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     7501 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     7561 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     7621 atgattacgc caagctctag ctagaggtcg accaattctc atgtttgaca gcttatcatc
     7681 gcagatccgg gcaacgttgt tgccattgct gcaggcgcag aactggtagg tatggaagat
     7741 ctatacattg aatcaatatt ggcaattagc catattagtc attggttata tagcataaat
     7801 caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat
     7861 tggctcatgt ccaatatgac cgccatgttg acattgatta ttgactagtt attaatagta
     7921 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     7981 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     8041 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     8101 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat
     8161 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga
     8221 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     8281 ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     8341 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     8401 tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     8461 tataagcaga gctcgtttag tgaaccgtca gatcctcact ctcttccgca tcgctgtctg
     8521 cgagggccag ctgttgggct cgcggttgag gacaaactct tcgcggtctt tccagtactc
     8581 ttggatcgga aacccgtcgg cctccgaacg gtactccgcc accgagggac ctgagcgagt
     8641 ccgcatcgac cggatcggaa aacct
//
