LOCUS       sequence-224487-        9446 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 9446)
  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 9446)
  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: PSOP7-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9446
                     /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             2980..3765
                     /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             3772..3837
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3844..3861
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3897..3952
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3951..3970)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6188..6206)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6274..6378
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6379..6447,6448..7239)
                     /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      7410..7998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7899..7918
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8152..8169
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8286..8307
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8322..8327,8328..8345,8346..8352)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8360..8376
                     /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     8365..8387
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8384..8400
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8384..8400
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8671..9050
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9051..9254
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9205..9225
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9251..9275
                     /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 ggccgccctg
      421 agcctgaagg gcatcaaaga gggcgaccct ccccggctga tgtacgacaa caacaaagtg
      481 aaccacggca acaccctgaa ggtgtccaag gacgagctga tgaagcggat cgtgcaggac
      541 gtgtcctacg atatcaacaa cttcatcaac gaccagtatg agaacaacaa taacaacaac
      601 aataataaca ataacaacaa caacaacatg cggcgggaca agaccagcga gaacgtgaaa
      661 gagctgcaga acgacgacac cgtgtacatc aagtccctgc agaacaagcc cctgctgaag
      721 atgaacaaca aggaaaaaaa caacaagatc gagatgaagg acagcaagaa gggcaactac
      781 aacaagtaca agatcaagca caacaacaag gcctacgacg gcggcatcag caacagcaga
      841 gagcagaaca acgacgagaa ctacatgaga gaggaacagc ggggcaacat ggccttcaag
      901 ctgaacaacc tgaacaagaa gaagtcccgg atcaagaaca ataagaacgg caagaaagag
      961 aacaatatcg acattaacaa caatatgatc aagatcaaca acatcaagaa ggacatcccc
     1021 aagatcgaca acgacaacaa caacaacaat aacaatgata acaacaataa taacaacaac
     1081 gataacaaca acaacaacaa taacgacaac aataacaata acaacaatgt ggacggcgac
     1141 gacaacaccg ccatccagaa caagggcccc accaatatcg ccagcaccaa cgagcacaag
     1201 aacatcaaca caaacaatca gaacaaacag aagctgcact ccgagaacca ccacaaaaac
     1261 aagctgaccg agacaagcca caacggcgac gtgatcgaga tcagaatcat cagcgacccc
     1321 ggcttcgtga agtccaacca caagaaaacc gagaagaaca agaaatacga catgaaggaa
     1381 aaggacctgc tgcgcatcaa gaacgggaag cggaacaaag aactgatggg cgtgcccgac
     1441 aagatgagca agcgccgcga ggtggaacgc gagaaggacc acatcaaggg caattctaaa
     1501 aacaagaata agaagaagaa taaaaacaaa aagaaaaaca agaacaagaa aaaaaacaag
     1561 aacaaaaact tcgccatcaa gaccaacgat gagtctaaga aaaataagat cagctacatc
     1621 cagatgtaca ccgacaagaa cggcgccgac gagtacagcc ccaagctgaa tgacgacgag
     1681 acattcgacg aggatatgta cgagggcaat gacgagatcg acagcaatat ggccattaag
     1741 aagatcaacg aggataagaa cctgctcgag ttcggcgacg agggcgtgaa ggatatcggc
     1801 tccaccgaga acgagatgat caaaaacaag aagaacatca tcaagaacag ccggaaggtg
     1861 tacaacctca acctgtacaa ctgcgatctg atcgacgact gggacctgaa ccacgagtac
     1921 atcgacgaag agggcaagct gatcaagctg agcggctacg tgttccagaa catcgtgtcc
     1981 accgacagca tgcccaccgt gaatatcgcc gactggaagc tgcggggcag ctgctcctac
     2041 gacaactaca tctgtggcgc cctgaattac atgaataaca tctacagcaa gggcgaccgc
     2101 gtgatcttcg acggcaagat ctacgaggcc accgccgagg tgtacgagac acccaaagtg
     2161 atggaaaacc tgtggatcga aaagaccaag gactgctacg acttcggcgc gccgtcgacc
     2221 tccatcacgg cctataagag tgagggggag tcagcggagt tctccttccc actcaacctt
     2281 ggagaggaaa gcctgcaggg agagttgaga tggaaggcag agaaggctcc ttcttcccag
     2341 tcctggatca ccttctccct aaagaaccaa aaggtgtctg tgcagaagtc tactagcaac
     2401 cccaagttcc agctgtccga aacgctccca ctcacccttc agatacccca ggtctccctt
     2461 cagtttgctg gttctggcaa cctgaccctg actctggaca gagggatact gtatcaggaa
     2521 gtgaacctgg tggtgatgaa agtgactcag cccgacagca acactttgac ctgtgaggtg
     2581 atgggaccca cctcacccaa gatgagactg atcttgaagc aggagaatca ggaggccagg
     2641 gtctccaggc aggagaaagt gattcaagtg caggcccctg aagcaggggt gtggcaatgt
     2701 ctactgagtg aaggtgaaga ggtcaagatg gactccaaga tccaggtttt atccaaaggg
     2761 ttgaattccg gcggcggctc cggaggcggt accggtggcg gcgacctagc cccacagatg
     2821 cttcgagaac tccaggagac taatgcggcg ctgcaagacg tgagagagct cttgcgacag
     2881 caggtcaagg agatcacctt cctgaagaat acggtgatgg aatgtgacgc ctgcggtcta
     2941 gacagaaacc tgcctccact agcacctctg ggcccacacc cagaaacgct ggtgaaagta
     3001 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3061 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3121 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3181 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3241 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3301 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3361 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3421 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3481 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3541 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3601 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3661 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3721 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggggccc tgactacaaa
     3781 gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga tgacaagggc
     3841 cctcaccacc accatcacca ctaataataa ggatcccccg acctcgacct ctggctaata
     3901 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     3961 gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag ctagaggatc
     4021 gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc ttcgcggggc
     4081 agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct gttccacatg
     4141 tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca tccttataaa
     4201 tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact ttgcagtctg
     4261 tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct tacaccaatg
     4321 ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca ccaacgtcaa
     4381 tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta gcaatagtgt
     4441 ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt agtagtatat
     4501 actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc atatgctatc
     4561 gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc atgagctgtc
     4621 acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt tagtcacaag
     4681 ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg cctgcttctt
     4741 cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa ggtgtatgtg
     4801 aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga cggggggttc
     4861 agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg gcaataaata
     4921 ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg gggtggggac
     4981 aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca acacataatc
     5041 ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc aagacaggtg
     5101 aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc gacagcagcg
     5161 gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg ccaatggggc
     5221 ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc
     5281 agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat aacttggctg
     5341 attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac taatggcacc
     5401 ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat tgctgcgatc
     5461 tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt tggtcctcat
     5521 attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca tatactaccc
     5581 aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat
     5641 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     5701 ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     5761 ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctaat
     5821 agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata ctacccaaat
     5881 atctggatag catatgctat cctaatctat atctgggtag catatgctat cctaatctat
     5941 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     6001 atctgggtag tatatgctat cctaatttat atctgggtag cataggctat cctaatctat
     6061 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt
     6121 atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt ttcttgaaga
     6181 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     6241 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6301 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6361 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6421 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6481 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6541 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6601 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     6661 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6721 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6781 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6841 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6901 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     6961 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     7021 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7081 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7141 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7201 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7261 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7321 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7381 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7441 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7501 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7561 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7621 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7681 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7741 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7801 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7861 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7921 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7981 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8041 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     8101 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     8161 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     8221 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     8281 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     8341 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     8401 catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac agcttatcat
     8461 cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag gtatggaaga
     8521 tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa
     8581 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
     8641 ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt tattaatagt
     8701 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     8761 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     8821 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     8881 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta
     8941 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg
     9001 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     9061 tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     9121 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     9181 gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     9241 atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc atcgctgtct
     9301 gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct ttccagtact
     9361 cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga cctgagcgag
     9421 tccgcatcga ccggatcgga aaacct
//
