LOCUS       sequence-224484-        9041 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 9041)
  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 9041)
  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_0620000-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9041
                     /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             2575..3360
                     /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             3367..3432
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3439..3456
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3492..3547
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3546..3565)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5783..5801)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5869..5973
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5974..6042,6043..6834)
                     /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      7005..7593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7494..7513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7747..7764
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7881..7902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7917..7922,7923..7940,7941..7947)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7955..7971
                     /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     7960..7982
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7979..7995
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7979..7995
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8266..8645
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8646..8849
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8800..8820
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8846..8870
                     /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 gacgatggac agagacacgg cgaggacagc gacagcatga acaagtacaa ctacatctac
      481 cggcccaacg ccgacaacgg catccacaac aacctgcggc ccagcagcta catcgacatg
      541 gaacacattg ccaagggccg ggacaacaca gccggcctga gagagaacga gctgttcgac
      601 atccagaacg acgagaacag caacatgggc cccgaccagg aaaaagtcat caccctgagc
      661 gacaagatct acatcgataa gatcaagaac atcatcgagt accggaagtg gcgggccaag
      721 agcaagtaca gaagccccgc caagcagccc gaagaggaca tgttcgtgga aggctacatc
      781 cccatcaagc ggcgggacga caacctgctg cctcacgaga aggccgagaa ggacttcgag
      841 gaaatcatcc agaaatacgt ggacctcgag aacaagctgc agaagcagaa agagctggaa
      901 gaggccgagc ggcggaagaa gaagctggac gacgagcaga agcggcagaa ggatctcgag
      961 gacaccaacc ggaaagagaa tgacgaggaa cagagctaca agaagctcga ggacctggaa
     1021 ctggaaaaca tcgacaccaa gatcaacaac aacgacgcct acgacgtgga aaacggcttc
     1081 aacgacgaca acaagctgta cacaaagatt aacaagatcg tcagccaggt gtcccagaac
     1141 aatgagctgt acgtgaacat catgaagtac atcacactgg tctacaccag ccacgtggac
     1201 atcaagcagg acgacttcac caacggcgcc atcagcatct ttctgacctt cgagaacccc
     1261 aagatgggca acgacctggc caatatcaat atcgccttct tcgccagcga gcagcggacc
     1321 ggcaccaacg aggtggccga ggccagaagc aactacctga ccaactacaa catgaatgag
     1381 ctgctcgagg ataacgccgc tggccggctg ctgagccagg acgagtacat caaagaactc
     1441 gtgaagtaca accacgccat ctccagcagc aacaaagaga acctgaagaa cgagaactac
     1501 gacgtcaaca tgaacattgc taagatcctg cagtatatca tcgacaacga catcaacctg
     1561 gtggacaact tcatcacctt caacagagcc aacgagcagg aagtgaccat cagcaagctg
     1621 gatgactttc tgcacgagtg ctccaaagtg aagcccgagg atatcaagaa ggatgagatc
     1681 cagaagatca tcaagaccat caacaagaaa gagcagatta tcaaagaagt gaagaacagc
     1741 ctgatcaaga aagacatcga gaagtgcaag ctctacacca ccatcctgat gttcggcagc
     1801 ggcgcgccgt cgacctccat cacggcctat aagagtgagg gggagtcagc ggagttctcc
     1861 ttcccactca accttggaga ggaaagcctg cagggagagt tgagatggaa ggcagagaag
     1921 gctccttctt cccagtcctg gatcaccttc tccctaaaga accaaaaggt gtctgtgcag
     1981 aagtctacta gcaaccccaa gttccagctg tccgaaacgc tcccactcac ccttcagata
     2041 ccccaggtct cccttcagtt tgctggttct ggcaacctga ccctgactct ggacagaggg
     2101 atactgtatc aggaagtgaa cctggtggtg atgaaagtga ctcagcccga cagcaacact
     2161 ttgacctgtg aggtgatggg acccacctca cccaagatga gactgatctt gaagcaggag
     2221 aatcaggagg ccagggtctc caggcaggag aaagtgattc aagtgcaggc ccctgaagca
     2281 ggggtgtggc aatgtctact gagtgaaggt gaagaggtca agatggactc caagatccag
     2341 gttttatcca aagggttgaa ttccggcggc ggctccggag gcggtaccgg tggcggcgac
     2401 ctagccccac agatgcttcg agaactccag gagactaatg cggcgctgca agacgtgaga
     2461 gagctcttgc gacagcaggt caaggagatc accttcctga agaatacggt gatggaatgt
     2521 gacgcctgcg gtctagacag aaacctgcct ccactagcac ctctgggccc acacccagaa
     2581 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2641 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2701 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     2761 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2821 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2881 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2941 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3001 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3061 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3121 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3181 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3241 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3301 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3361 ggccctgact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     3421 gacgatgaca agggccctca ccaccaccat caccactaat aataaggatc ccccgacctc
     3481 gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     3541 tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc
     3601 tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc
     3661 ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg
     3721 gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt
     3781 tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc
     3841 agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa
     3901 gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg
     3961 ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
     4021 cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc
     4081 cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg
     4141 gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc
     4201 accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac
     4261 cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
     4321 cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac
     4381 agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat
     4441 ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc
     4501 cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat
     4561 ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
     4621 gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag
     4681 ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg
     4741 acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct
     4801 ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga
     4861 gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
     4921 gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa
     4981 accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc
     5041 gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg
     5101 gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg
     5161 tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
     5221 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     5281 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     5341 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     5401 atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc
     5461 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
     5521 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     5581 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     5641 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     5701 gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga
     5761 gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5821 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5881 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5941 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     6001 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     6061 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6121 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6181 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     6241 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6301 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6361 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6421 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6481 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     6541 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6601 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6661 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6721 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6781 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6841 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6901 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6961 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7021 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7081 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7141 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7201 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7261 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7321 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7381 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7441 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7501 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7561 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7621 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7681 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     7741 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     7801 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     7861 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7921 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7981 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt
     8041 ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact
     8101 ggtaggtatg gaagatctat acattgaatc aatattggca attagccata ttagtcattg
     8161 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
     8221 atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttgacat tgattattga
     8281 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     8341 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     8401 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     8461 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     8521 caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     8581 acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     8641 ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg
     8701 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     8761 gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg
     8821 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc ctcactctct
     8881 tccgcatcgc tgtctgcgag ggccagctgt tgggctcgcg gttgaggaca aactcttcgc
     8941 ggtctttcca gtactcttgg atcggaaacc cgtcggcctc cgaacggtac tccgccaccg
     9001 agggacctga gcgagtccgc atcgaccgga tcggaaaacc t
//
