LOCUS       sequence-208702-        8827 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 8827)
  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 8827)
  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..8827
                     /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    3278..3333
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3332..3351)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5569..5587)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5655..5759
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5760..5828,5829..6620)
                     /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      6791..7379
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7280..7299
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7533..7550
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7667..7688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7703..7708,7709..7726,7727..7733)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7741..7757
                     /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     7746..7768
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7765..7781
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7765..7781
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8052..8431
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8432..8635
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8586..8606
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8632..8656
                     /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 taattaatta aggatccccc gacctcgacc tctggctaat aaaggaaatt tattttcatt
     3301 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     3361 catttggtcg agatccctcg gagatctcta gctagaggat cgatccccgc cccggacgaa
     3421 ctaaacctga ctacgacatc tctgcccctt cttcgcgggg cagtgcatgt aatcccttca
     3481 gttggttggt acaacttgcc aactgggccc tgttccacat gtgacacggg gggggaccaa
     3541 acacaaaggg gttctctgac tgtagttgac atccttataa atggatgtgc acatttgcca
     3601 acactgagtg gctttcatcc tggagcagac tttgcagtct gtggactgca acacaacatt
     3661 gcctttatgt gtaactcttg gctgaagctc ttacaccaat gctgggggac atgtacctcc
     3721 caggggccca ggaagactac gggaggctac accaacgtca atcagagggg cctgtgtagc
     3781 taccgataag cggaccctca agagggcatt agcaatagtg tttataaggc ccccttgtta
     3841 accctaaacg ggtagcatat gcttcccggg tagtagtata tactatccag actaacccta
     3901 attcaatagc atatgttacc caacgggaag catatgctat cgaattaggg ttagtaaaag
     3961 ggtcctaagg aacagcgata tctcccaccc catgagctgt cacggtttta tttacatggg
     4021 gtcaggattc cacgagggta gtgaaccatt ttagtcacaa gggcagtggc tgaagatcaa
     4081 ggagcgggca gtgaactctc ctgaatcttc gcctgcttct tcattctcct tcgtttagct
     4141 aatagaataa ctgctgagtt gtgaacagta aggtgtatgt gaggtgctcg aaaacaaggt
     4201 ttcaggtgac gcccccagaa taaaatttgg acggggggtt cagtggtggc attgtgctat
     4261 gacaccaata taaccctcac aaaccccttg ggcaataaat actagtgtag gaatgaaaca
     4321 ttctgaatat ctttaacaat agaaatccat ggggtgggga caagccgtaa agactggatg
     4381 tccatctcac acgaatttat ggctatgggc aacacataat cctagtgcaa tatgatactg
     4441 gggttattaa gatgtgtccc aggcagggac caagacaggt gaaccatgtt gttacactct
     4501 atttgtaaca aggggaaaga gagtggacgc cgacagcagc ggactccact ggttgtctct
     4561 aacacccccg aaaattaaac ggggctccac gccaatgggg cccataaaca aagacaagtg
     4621 gccactcttt tttttgaaat tgtggagtgg gggcacgcgt cagcccccac acgccgccct
     4681 gcggttttgg actgtaaaat aagggtgtaa taacttggct gattgtaacc ccgctaacca
     4741 ctgcggtcaa accacttgcc cacaaaacca ctaatggcac cccggggaat acctgcataa
     4801 gtaggtgggc gggccaagat aggggcgcga ttgctgcgat ctggaggaca aattacacac
     4861 acttgcgcct gagcgccaag cacagggttg ttggtcctca tattcacgag gtcgctgaga
     4921 gcacggtggg ctaatgttgc catgggtagc atatactacc caaatatctg gatagcatat
     4981 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     5041 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     5101 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     5161 gctatcctaa tctgtatccg ggtagcatat gctatcctaa tagagattag ggtagtatat
     5221 gctatcctaa tttatatctg ggtagcatat actacccaaa tatctggata gcatatgcta
     5281 tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta gcataggcta
     5341 tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta
     5401 tcctaattta tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta
     5461 tcctaatcta tatctgggta gtatatgcta tcctaatctg tatccgggta gcatatgcta
     5521 tcctcatgat aagctgtcaa acatgagaat tttcttgaag acgaaagggc ctcgtgatac
     5581 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5641 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5701 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5761 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5821 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5881 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5941 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6001 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6061 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6121 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6181 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6241 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6301 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6361 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6421 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6481 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6541 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6601 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6661 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6721 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6781 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6841 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6901 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6961 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7021 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7081 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7141 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7201 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7261 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7321 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7381 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7441 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     7501 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     7561 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     7621 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     7681 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     7741 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
     7801 agctagaggt cgaccaattc tcatgtttga cagcttatca tcgcagatcc gggcaacgtt
     7861 gttgccattg ctgcaggcgc agaactggta ggtatggaag atctatacat tgaatcaata
     7921 ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg gctattggcc
     7981 attgcatacg ttgtatctat atcataatat gtacatttat attggctcat gtccaatatg
     8041 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     8101 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     8161 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     8221 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     8281 ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa
     8341 atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta
     8401 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acaccaatgg
     8461 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     8521 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata accccgcccc
     8581 gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
     8641 agtgaaccgt cagatcctca ctctcttccg catcgctgtc tgcgagggcc agctgttggg
     8701 ctcgcggttg aggacaaact cttcgcggtc tttccagtac tcttggatcg gaaacccgtc
     8761 ggcctccgaa cggtactccg ccaccgaggg acctgagcga gtccgcatcg accggatcgg
     8821 aaaacct
//
