LOCUS       sequence-224709-        9170 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 9170)
  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 9170)
  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: PF3D7_1242000-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9170
                     /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             2704..3489
                     /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             3496..3561
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3568..3585
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3621..3676
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3675..3694)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5912..5930)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5998..6102
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6103..6171,6172..6963)
                     /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      7134..7722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7623..7642
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7876..7893
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8010..8031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8046..8051,8052..8069,8070..8076)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8084..8100
                     /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     8089..8111
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8108..8124
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8108..8124
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8395..8774
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8775..8978
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8929..8949
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8975..8999
                     /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 ggccgccgat
      421 aatgtgtgca agggcgccga cgtgatcgtg aacgagttct ggtccaagct gaacaacatc
      481 cggcacgggg acgtgatcct gatcaacatc aagaactact actgccccgc ctgcaaccgg
      541 tacatgaaca tctggaacga cgtcgagaag aagatcctga cctacgagaa gaacgtggcc
      601 ctgttcgtgt tcgactgcag ctgctacctg ctggtgccct actgccggta cttcgacgtg
      661 ctgtacttcc ccaccttccg gctgctgttc cccgtgtacg acaagatcaa cgacaaagag
      721 tacaagtaca tcaaccccgc cagcatgatc gagggcgaga agtacaacgg cgacctgctg
      781 ctggcctaca gagaggtgga acggatcgac aacatctacg acttcaagat cctcatcgag
      841 aagtatctgt gcaagaacgt gaacttcaac tacaacgacc tgcggatctg catgaacagc
      901 gcctacatcc tgcccgagaa gaaaaacaag aagaagaacg agatctacca cttcctgttc
      961 ggctacaaga acgacaagaa caaagaagag atcaaggaca gcgtgaacgt gatgaacgac
     1021 gactttatcg agaacaacta cgattacgaa gaaaacaaag aaaacgtcga gcggtggtcc
     1081 aaccacatca acttcaacaa ggacaatatc aagcacgaca tcatcatcgg catcctgttc
     1141 accctgaaga agcacatcag cctgggccgg gacatccaga aagagtatat cgagcccttc
     1201 ctggtcatgc tgagcatcgt gtccaacatc tataacgacc tggaagaggg cctgtacgat
     1261 atcatcaaca agctcaacag cttcacctac cccattaagt acgacgagtg gatcaagtac
     1321 accgagaaca tcaattacat caacgagtac aagctggact acaacaacaa taacaacaac
     1381 gacgccaacc tgatcagctt caagctgtgc gagcagaaca gcgtgctgtg cagctactgg
     1441 ctgctgtatc acaagatcag cgtgtactgc ctgcagcacg acaagcacaa ctacctgtac
     1501 tacatcgagg ccatcaccaa ctacgccaag aattacctga actgccagaa ctgcatcgac
     1561 cactttctga acgcccagaa gttctgctac tacggctact gcaacatcca cagcgccgag
     1621 agcttcatca tctttctgtg gcggatccac aacgccgtga ccctgcggtc tatgtacgat
     1681 ctgctgatga tcgataacaa cgccgccacc cctagcaaca atttcaacaa gaagcagttc
     1741 ctgaaccagg acatcgtgtt ccctaccctg aagcagtgca agaattgccg gaacgccctg
     1801 ggcttcacca agatcaccaa tcacgccctg agcaccttca agaacggcgc catcaccgac
     1861 cagagcttcg acgccatcga cagcttcagc gtgaagaatg tgctgcacta cctgatccac
     1921 atctacagcg gcgcgccgtc gacctccatc acggcctata agagtgaggg ggagtcagcg
     1981 gagttctcct tcccactcaa ccttggagag gaaagcctgc agggagagtt gagatggaag
     2041 gcagagaagg ctccttcttc ccagtcctgg atcaccttct ccctaaagaa ccaaaaggtg
     2101 tctgtgcaga agtctactag caaccccaag ttccagctgt ccgaaacgct cccactcacc
     2161 cttcagatac cccaggtctc ccttcagttt gctggttctg gcaacctgac cctgactctg
     2221 gacagaggga tactgtatca ggaagtgaac ctggtggtga tgaaagtgac tcagcccgac
     2281 agcaacactt tgacctgtga ggtgatggga cccacctcac ccaagatgag actgatcttg
     2341 aagcaggaga atcaggaggc cagggtctcc aggcaggaga aagtgattca agtgcaggcc
     2401 cctgaagcag gggtgtggca atgtctactg agtgaaggtg aagaggtcaa gatggactcc
     2461 aagatccagg ttttatccaa agggttgaat tccggcggcg gctccggagg cggtaccggt
     2521 ggcggcgacc tagccccaca gatgcttcga gaactccagg agactaatgc ggcgctgcaa
     2581 gacgtgagag agctcttgcg acagcaggtc aaggagatca ccttcctgaa gaatacggtg
     2641 atggaatgtg acgcctgcgg tctagacaga aacctgcctc cactagcacc tctgggccca
     2701 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2761 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2821 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2881 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2941 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3001 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3061 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3121 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3181 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3241 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3301 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3361 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3421 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3481 aagcattggg gccctgacta caaagaccat gacggtgatt ataaagatca tgacatcgat
     3541 tacaaggatg acgatgacaa gggccctcac caccaccatc accactaata ataaggatcc
     3601 cccgacctcg acctctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     3661 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttgg tcgagatccc
     3721 tcggagatct ctagctagag gatcgatccc cgccccggac gaactaaacc tgactacgac
     3781 atctctgccc cttcttcgcg gggcagtgca tgtaatccct tcagttggtt ggtacaactt
     3841 gccaactggg ccctgttcca catgtgacac ggggggggac caaacacaaa ggggttctct
     3901 gactgtagtt gacatcctta taaatggatg tgcacatttg ccaacactga gtggctttca
     3961 tcctggagca gactttgcag tctgtggact gcaacacaac attgccttta tgtgtaactc
     4021 ttggctgaag ctcttacacc aatgctgggg gacatgtacc tcccaggggc ccaggaagac
     4081 tacgggaggc tacaccaacg tcaatcagag gggcctgtgt agctaccgat aagcggaccc
     4141 tcaagagggc attagcaata gtgtttataa ggcccccttg ttaaccctaa acgggtagca
     4201 tatgcttccc gggtagtagt atatactatc cagactaacc ctaattcaat agcatatgtt
     4261 acccaacggg aagcatatgc tatcgaatta gggttagtaa aagggtccta aggaacagcg
     4321 atatctccca ccccatgagc tgtcacggtt ttatttacat ggggtcagga ttccacgagg
     4381 gtagtgaacc attttagtca caagggcagt ggctgaagat caaggagcgg gcagtgaact
     4441 ctcctgaatc ttcgcctgct tcttcattct ccttcgttta gctaatagaa taactgctga
     4501 gttgtgaaca gtaaggtgta tgtgaggtgc tcgaaaacaa ggtttcaggt gacgccccca
     4561 gaataaaatt tggacggggg gttcagtggt ggcattgtgc tatgacacca atataaccct
     4621 cacaaacccc ttgggcaata aatactagtg taggaatgaa acattctgaa tatctttaac
     4681 aatagaaatc catggggtgg ggacaagccg taaagactgg atgtccatct cacacgaatt
     4741 tatggctatg ggcaacacat aatcctagtg caatatgata ctggggttat taagatgtgt
     4801 cccaggcagg gaccaagaca ggtgaaccat gttgttacac tctatttgta acaaggggaa
     4861 agagagtgga cgccgacagc agcggactcc actggttgtc tctaacaccc ccgaaaatta
     4921 aacggggctc cacgccaatg gggcccataa acaaagacaa gtggccactc ttttttttga
     4981 aattgtggag tgggggcacg cgtcagcccc cacacgccgc cctgcggttt tggactgtaa
     5041 aataagggtg taataacttg gctgattgta accccgctaa ccactgcggt caaaccactt
     5101 gcccacaaaa ccactaatgg caccccgggg aatacctgca taagtaggtg ggcgggccaa
     5161 gataggggcg cgattgctgc gatctggagg acaaattaca cacacttgcg cctgagcgcc
     5221 aagcacaggg ttgttggtcc tcatattcac gaggtcgctg agagcacggt gggctaatgt
     5281 tgccatgggt agcatatact acccaaatat ctggatagca tatgctatcc taatctatat
     5341 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     5401 ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat
     5461 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat
     5521 ccgggtagca tatgctatcc taatagagat tagggtagta tatgctatcc taatttatat
     5581 ctgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg
     5641 gtagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg
     5701 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg
     5761 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     5821 gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctcat gataagctgt
     5881 caaacatgag aattttcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5941 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     6001 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6061 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6121 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6181 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6241 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6301 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     6361 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6421 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6481 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6541 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6601 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     6661 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6721 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6781 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6841 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6901 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6961 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7021 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7081 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7141 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7201 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7261 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7321 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7381 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7441 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7501 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7561 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7621 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7681 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7741 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7801 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     7861 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     7921 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     7981 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     8041 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     8101 tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgaccaa
     8161 ttctcatgtt tgacagctta tcatcgcaga tccgggcaac gttgttgcca ttgctgcagg
     8221 cgcagaactg gtaggtatgg aagatctata cattgaatca atattggcaa ttagccatat
     8281 tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc
     8341 tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca tgttgacatt
     8401 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     8461 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     8521 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     8581 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     8641 atcatatgcc aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     8701 atgcccagta catgacctta cgggactttc ctacttggca gtacatctac gtattagtca
     8761 tcgctattac catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg
     8821 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     8881 aaaatcaacg ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg
     8941 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcc
     9001 tcactctctt ccgcatcgct gtctgcgagg gccagctgtt gggctcgcgg ttgaggacaa
     9061 actcttcgcg gtctttccag tactcttgga tcggaaaccc gtcggcctcc gaacggtact
     9121 ccgccaccga gggacctgag cgagtccgca tcgaccggat cggaaaacct
//
