LOCUS       sequence-224370-        8969 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 8969)
  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 8969)
  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: PSOP24-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8969
                     /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             2503..3288
                     /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             3295..3360
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3367..3384
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3420..3475
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3474..3493)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5711..5729)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5797..5901
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5902..5970,5971..6762)
                     /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      6933..7521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7422..7441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7675..7692
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7809..7830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7845..7850,7851..7868,7869..7875)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7883..7899
                     /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     7888..7910
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7907..7923
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7907..7923
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8194..8573
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8574..8777
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8728..8748
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8774..8798
                     /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 aacatcaacc tgaacgacga gggcaagaag aagcagagcg gcatcatcaa cgagaatgag
      481 gacggcaaga acaacaaggc caacaacaac aagaaggtgc agcacaacaa gatcaaccgg
      541 aagggcaaca acgccgtgac caagaaaacc gacgagcaca agaacgacgg cgacgccgag
      601 aacaagaaga atggcgagca caaaaatgat ggggacggga aaaacaagaa aaacggggaa
      661 cataagaacg atggggatgg aaagaacaaa aagatggatg agcacctgta cgacatcgac
      721 ggggagaata agaaaaatgg tgaacacctc tacgatgtgg acggcgagaa caaaaaaaac
      781 ggcgaacacc tgtatgatat cgatggcgaa aacaaaaaga acggggagca tctctacgac
      841 attgatggcg agaacaagaa aacagatgag cacctgtatg acgtggacgg cggcaaaaac
      901 aacatcctcg agtacaacaa catcgaccag cacaacgagt tccccgagag cctggaaaac
      961 gacaacagct acgacaagct gttcgacgac gtggaactgc gggacatcat ccacaacgag
     1021 aagttcttcg agaacctgaa gaacgtgaac aacaacgacg tgcacaactt tctgctggtg
     1081 gacaaagaga tggaacgccg caaagaggaa gagaagaaga agtcccggaa gctggaagag
     1141 gacgaagagg atgacgagga cgaggaagaa gataacgacg agaagtggaa agaggaaaat
     1201 aagaacaacg ctgccagcag cagctcctcc agcagcaaca acttcaacaa gaactacgac
     1261 atctacgaca accgggacct gtacaactac gataacctgg tggatatcac ccaggaagaa
     1321 cacatcaacg tccccaacat caacgatgag atcaacgagg gccggaagca cgacaaagag
     1381 gactttctga tctttgagaa gcggctgaac gaggccgagg accggaacga cgccagcgac
     1441 ggggataaca agatggaaga tctgatcaac aaccacttcg acgacagcgg cgacaacggc
     1501 gcctacgaga acatccaggt caaaaaagag ctgtacaaca aaaacgagaa gatcatcaag
     1561 ggccgggaca acaacctgaa tgatggcgac atccacatga acaaaaacaa tgccgccaac
     1621 aaactcaaca acaataacaa caacaacaac aatgacaaca atatgaacag cgagatgaag
     1681 tacaagacct tcaaagaaga gtacgaggaa cagatcctga gcaagcccgg cgcgccgtcg
     1741 acctccatca cggcctataa gagtgagggg gagtcagcgg agttctcctt cccactcaac
     1801 cttggagagg aaagcctgca gggagagttg agatggaagg cagagaaggc tccttcttcc
     1861 cagtcctgga tcaccttctc cctaaagaac caaaaggtgt ctgtgcagaa gtctactagc
     1921 aaccccaagt tccagctgtc cgaaacgctc ccactcaccc ttcagatacc ccaggtctcc
     1981 cttcagtttg ctggttctgg caacctgacc ctgactctgg acagagggat actgtatcag
     2041 gaagtgaacc tggtggtgat gaaagtgact cagcccgaca gcaacacttt gacctgtgag
     2101 gtgatgggac ccacctcacc caagatgaga ctgatcttga agcaggagaa tcaggaggcc
     2161 agggtctcca ggcaggagaa agtgattcaa gtgcaggccc ctgaagcagg ggtgtggcaa
     2221 tgtctactga gtgaaggtga agaggtcaag atggactcca agatccaggt tttatccaaa
     2281 gggttgaatt ccggcggcgg ctccggaggc ggtaccggtg gcggcgacct agccccacag
     2341 atgcttcgag aactccagga gactaatgcg gcgctgcaag acgtgagaga gctcttgcga
     2401 cagcaggtca aggagatcac cttcctgaag aatacggtga tggaatgtga cgcctgcggt
     2461 ctagacagaa acctgcctcc actagcacct ctgggcccac acccagaaac gctggtgaaa
     2521 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2581 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2641 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     2701 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2761 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2821 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2881 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2941 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3001 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3061 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3121 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3181 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3241 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattgggg ccctgactac
     3301 aaagaccatg acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag
     3361 ggccctcacc accaccatca ccactaataa taaggatccc ccgacctcga cctctggcta
     3421 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     3481 aaggacatat gggagggcaa atcatttggt cgagatccct cggagatctc tagctagagg
     3541 atcgatcccc gccccggacg aactaaacct gactacgaca tctctgcccc ttcttcgcgg
     3601 ggcagtgcat gtaatccctt cagttggttg gtacaacttg ccaactgggc cctgttccac
     3661 atgtgacacg gggggggacc aaacacaaag gggttctctg actgtagttg acatccttat
     3721 aaatggatgt gcacatttgc caacactgag tggctttcat cctggagcag actttgcagt
     3781 ctgtggactg caacacaaca ttgcctttat gtgtaactct tggctgaagc tcttacacca
     3841 atgctggggg acatgtacct cccaggggcc caggaagact acgggaggct acaccaacgt
     3901 caatcagagg ggcctgtgta gctaccgata agcggaccct caagagggca ttagcaatag
     3961 tgtttataag gcccccttgt taaccctaaa cgggtagcat atgcttcccg ggtagtagta
     4021 tatactatcc agactaaccc taattcaata gcatatgtta cccaacggga agcatatgct
     4081 atcgaattag ggttagtaaa agggtcctaa ggaacagcga tatctcccac cccatgagct
     4141 gtcacggttt tatttacatg gggtcaggat tccacgaggg tagtgaacca ttttagtcac
     4201 aagggcagtg gctgaagatc aaggagcggg cagtgaactc tcctgaatct tcgcctgctt
     4261 cttcattctc cttcgtttag ctaatagaat aactgctgag ttgtgaacag taaggtgtat
     4321 gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag aataaaattt ggacgggggg
     4381 ttcagtggtg gcattgtgct atgacaccaa tataaccctc acaaacccct tgggcaataa
     4441 atactagtgt aggaatgaaa cattctgaat atctttaaca atagaaatcc atggggtggg
     4501 gacaagccgt aaagactgga tgtccatctc acacgaattt atggctatgg gcaacacata
     4561 atcctagtgc aatatgatac tggggttatt aagatgtgtc ccaggcaggg accaagacag
     4621 gtgaaccatg ttgttacact ctatttgtaa caaggggaaa gagagtggac gccgacagca
     4681 gcggactcca ctggttgtct ctaacacccc cgaaaattaa acggggctcc acgccaatgg
     4741 ggcccataaa caaagacaag tggccactct tttttttgaa attgtggagt gggggcacgc
     4801 gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt aataacttgg
     4861 ctgattgtaa ccccgctaac cactgcggtc aaaccacttg cccacaaaac cactaatggc
     4921 accccgggga atacctgcat aagtaggtgg gcgggccaag ataggggcgc gattgctgcg
     4981 atctggagga caaattacac acacttgcgc ctgagcgcca agcacagggt tgttggtcct
     5041 catattcacg aggtcgctga gagcacggtg ggctaatgtt gccatgggta gcatatacta
     5101 cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat aggctatcct
     5161 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     5221 aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     5281 aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct
     5341 aatagagatt agggtagtat atgctatcct aatttatatc tgggtagcat atactaccca
     5401 aatatctgga tagcatatgc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     5461 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     5521 tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc
     5581 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc
     5641 tgtatccggg tagcatatgc tatcctcatg ataagctgtc aaacatgaga attttcttga
     5701 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5761 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5821 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5881 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5941 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6001 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6061 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6121 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     6181 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6241 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6301 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6361 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6421 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     6481 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6541 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6601 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6661 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6721 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6781 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6841 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6901 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6961 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7021 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7081 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7141 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7201 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7261 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7321 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7381 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7441 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7501 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7561 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7621 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     7681 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     7741 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     7801 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     7861 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     7921 gaccatgatt acgccaagct ctagctagag gtcgaccaat tctcatgttt gacagcttat
     7981 catcgcagat ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg taggtatgga
     8041 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
     8101 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
     8161 atattggctc atgtccaata tgaccgccat gttgacattg attattgact agttattaat
     8221 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     8281 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     8341 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     8401 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
     8461 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
     8521 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     8581 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     8641 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     8701 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
     8761 tctatataag cagagctcgt ttagtgaacc gtcagatcct cactctcttc cgcatcgctg
     8821 tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt
     8881 actcttggat cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc
     8941 gagtccgcat cgaccggatc ggaaaacct
//
