LOCUS       sequence-224283-        8825 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 8825)
  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 8825)
  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: SSP3-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8825
                     /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             2359..3144
                     /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             3151..3216
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3223..3240
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3276..3331
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3330..3349)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5567..5585)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5653..5757
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5758..5826,5827..6618)
                     /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      6789..7377
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7278..7297
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7531..7548
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7665..7686
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7701..7706,7707..7724,7725..7731)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7739..7755
                     /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     7744..7766
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7763..7779
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7763..7779
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8050..8429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8430..8633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8584..8604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8630..8654
                     /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 ggccgccgag
      421 ggctacatct gcgacttcag cagcgagaag tacaacatct acttcgacga gcagaaggac
      481 gacgtcatct gctaccacga gatctacgag ggcgacagca tcggcatcat catccccaga
      541 tacaaagagg gcaacgagaa catctacatc aagaccaact gcttcaacga ggtgtccctg
      601 gaaagcagcg gcaacaacgc cctgagcatc tacgacatct tcaacgacga cgagatcacc
      661 ctgtacgacc aggacagcat cttctacacc gagtacatca gcagcatcct gcacatcaag
      721 aaagtgaaga agaacaccta cctgcactgc gtgttcgaga gcaaggacgt gaacggcctg
      781 accacccaca agggaatcgc caagatcacc attaagaacg tgaagcacaa tgagaacaag
      841 aagctgatca acaacgacgt ggacaacaat gccgccctgt ccttcaagaa gaaactgacc
      901 aacaagctgg tcatcgacct gctgaacaag atcgacatca tcaaggatga caagctgaac
      961 aaccagtaca acaacgatga caacaagttc ctggtggaag ccaagcccgg cgacatcctg
     1021 tacctgctgg gcgtgaagcg gaagaacaaa aagtacctgt acgtggacca ggactgccct
     1081 ctgcacttcg agcggatcgg cagcatctac aagtacatct tccccatcat caacgagaaa
     1141 gaggaaatct acaactgcgc cctgtatgtg gacgacgacg tgtccaccaa caagaaaatc
     1201 gtgggcaacc tgaagatcac cttcgtgtcc aagaaccccg ccatcaacga catcaacatt
     1261 gacgtgctga agaagtacat ccacagcgac attgagcaga agctgtacaa gctgctgtac
     1321 ggcgagatca gcaacagcgt gaacaaatac gccgagcaga acgtggacat taacaacgcc
     1381 gacatcagcg ccatccagtt catgaatctg aacagcgaca actgcaacaa caatgagtgc
     1441 atcgagttct tcaacaacgc tgcctgcagc agctactgcg gcaagggcta ccggctgctg
     1501 aacggcaaca atcatatgta cgatgagcag atggtggtgc cctgctacaa cggcgagtgc
     1561 aagcccgagg acgagatcga acctggcgcg ccgtcgacct ccatcacggc ctataagagt
     1621 gagggggagt cagcggagtt ctccttccca ctcaaccttg gagaggaaag cctgcaggga
     1681 gagttgagat ggaaggcaga gaaggctcct tcttcccagt cctggatcac cttctcccta
     1741 aagaaccaaa aggtgtctgt gcagaagtct actagcaacc ccaagttcca gctgtccgaa
     1801 acgctcccac tcacccttca gataccccag gtctcccttc agtttgctgg ttctggcaac
     1861 ctgaccctga ctctggacag agggatactg tatcaggaag tgaacctggt ggtgatgaaa
     1921 gtgactcagc ccgacagcaa cactttgacc tgtgaggtga tgggacccac ctcacccaag
     1981 atgagactga tcttgaagca ggagaatcag gaggccaggg tctccaggca ggagaaagtg
     2041 attcaagtgc aggcccctga agcaggggtg tggcaatgtc tactgagtga aggtgaagag
     2101 gtcaagatgg actccaagat ccaggtttta tccaaagggt tgaattccgg cggcggctcc
     2161 ggaggcggta ccggtggcgg cgacctagcc ccacagatgc ttcgagaact ccaggagact
     2221 aatgcggcgc tgcaagacgt gagagagctc ttgcgacagc aggtcaagga gatcaccttc
     2281 ctgaagaata cggtgatgga atgtgacgcc tgcggtctag acagaaacct gcctccacta
     2341 gcacctctgg gcccacaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     2401 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     2461 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     2521 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     2581 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     2641 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     2701 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2761 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2821 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2881 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2941 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3001 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3061 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3121 ggtgcctcac tgattaagca ttggggccct gactacaaag accatgacgg tgattataaa
     3181 gatcatgaca tcgattacaa ggatgacgat gacaagggcc ctcaccacca ccatcaccac
     3241 taataataag gatcccccga cctcgacctc tggctaataa aggaaattta ttttcattgc
     3301 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     3361 tttggtcgag atccctcgga gatctctagc tagaggatcg atccccgccc cggacgaact
     3421 aaacctgact acgacatctc tgccccttct tcgcggggca gtgcatgtaa tcccttcagt
     3481 tggttggtac aacttgccaa ctgggccctg ttccacatgt gacacggggg gggaccaaac
     3541 acaaaggggt tctctgactg tagttgacat ccttataaat ggatgtgcac atttgccaac
     3601 actgagtggc tttcatcctg gagcagactt tgcagtctgt ggactgcaac acaacattgc
     3661 ctttatgtgt aactcttggc tgaagctctt acaccaatgc tgggggacat gtacctccca
     3721 ggggcccagg aagactacgg gaggctacac caacgtcaat cagaggggcc tgtgtagcta
     3781 ccgataagcg gaccctcaag agggcattag caatagtgtt tataaggccc ccttgttaac
     3841 cctaaacggg tagcatatgc ttcccgggta gtagtatata ctatccagac taaccctaat
     3901 tcaatagcat atgttaccca acgggaagca tatgctatcg aattagggtt agtaaaaggg
     3961 tcctaaggaa cagcgatatc tcccacccca tgagctgtca cggttttatt tacatggggt
     4021 caggattcca cgagggtagt gaaccatttt agtcacaagg gcagtggctg aagatcaagg
     4081 agcgggcagt gaactctcct gaatcttcgc ctgcttcttc attctccttc gtttagctaa
     4141 tagaataact gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa aacaaggttt
     4201 caggtgacgc ccccagaata aaatttggac ggggggttca gtggtggcat tgtgctatga
     4261 caccaatata accctcacaa accccttggg caataaatac tagtgtagga atgaaacatt
     4321 ctgaatatct ttaacaatag aaatccatgg ggtggggaca agccgtaaag actggatgtc
     4381 catctcacac gaatttatgg ctatgggcaa cacataatcc tagtgcaata tgatactggg
     4441 gttattaaga tgtgtcccag gcagggacca agacaggtga accatgttgt tacactctat
     4501 ttgtaacaag gggaaagaga gtggacgccg acagcagcgg actccactgg ttgtctctaa
     4561 cacccccgaa aattaaacgg ggctccacgc caatggggcc cataaacaaa gacaagtggc
     4621 cactcttttt tttgaaattg tggagtgggg gcacgcgtca gcccccacac gccgccctgc
     4681 ggttttggac tgtaaaataa gggtgtaata acttggctga ttgtaacccc gctaaccact
     4741 gcggtcaaac cacttgccca caaaaccact aatggcaccc cggggaatac ctgcataagt
     4801 aggtgggcgg gccaagatag gggcgcgatt gctgcgatct ggaggacaaa ttacacacac
     4861 ttgcgcctga gcgccaagca cagggttgtt ggtcctcata ttcacgaggt cgctgagagc
     4921 acggtgggct aatgttgcca tgggtagcat atactaccca aatatctgga tagcatatgc
     4981 tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     5041 tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc
     5101 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     5161 tatcctaatc tgtatccggg tagcatatgc tatcctaata gagattaggg tagtatatgc
     5221 tatcctaatt tatatctggg tagcatatac tacccaaata tctggatagc atatgctatc
     5281 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagc ataggctatc
     5341 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     5401 ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     5461 ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc
     5521 ctcatgataa gctgtcaaac atgagaattt tcttgaagac gaaagggcct cgtgatacgc
     5581 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5641 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5701 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5761 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5821 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5881 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5941 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6001 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6061 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6121 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6181 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6241 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6301 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6361 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6421 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6481 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6541 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6601 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6661 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6721 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6781 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6841 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6901 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6961 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7021 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7081 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7141 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7201 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7261 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7321 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7381 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7441 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7501 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7561 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7621 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7681 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7741 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag
     7801 ctagaggtcg accaattctc atgtttgaca gcttatcatc gcagatccgg gcaacgttgt
     7861 tgccattgct gcaggcgcag aactggtagg tatggaagat ctatacattg aatcaatatt
     7921 ggcaattagc catattagtc attggttata tagcataaat caatattggc tattggccat
     7981 tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac
     8041 cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     8101 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     8161 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     8221 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     8281 cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat
     8341 ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca
     8401 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc
     8461 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     8521 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaataac cccgccccgt
     8581 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
     8641 tgaaccgtca gatcctcact ctcttccgca tcgctgtctg cgagggccag ctgttgggct
     8701 cgcggttgag gacaaactct tcgcggtctt tccagtactc ttggatcgga aacccgtcgg
     8761 cctccgaacg gtactccgcc accgagggac ctgagcgagt ccgcatcgac cggatcggaa
     8821 aacct
//
