LOCUS       sequence-208753-        8209 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 8209)
  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 8209)
  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: SOAP-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8209
                     /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             1741..2526
                     /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             2533..2598
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2605..2622
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2660..2715
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2714..2733)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4951..4969)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5037..5141
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5142..5210,5211..6002)
                     /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      6173..6761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6662..6681
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6915..6932
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7049..7070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7085..7090,7091..7108,7109..7115)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7123..7139
                     /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     7128..7150
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7147..7163
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7147..7163
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7434..7813
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7814..8017
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7968..7988
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8014..8038
                     /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 ggccgccaaa
      421 ggcgtgcgga acaaggccga gaacagagcc ccttacgtgc actaccacaa caacgccaac
      481 atcaagaact acgccgccgt ggacatcttc agccccaaga aaaccggcaa agaatgcatc
      541 aagtgcctgc ccgacaactt ctgcgagtgc gagtgctcct gcaagaacaa ggctggcttc
      601 agcatgaagt accggcacgc cagcaagggc tccaagggct acagcaagaa gatgaccaag
      661 accagcagcc ggtcccggca gtgcaacgac ctgcccctgg aaaagagcga gcaggaaacc
      721 gagcccgagc tggaacctga ggtggaaccc gaggtcgagc ctgaagtcga acccgaagtc
      781 gaaccagagg tcgagcccga ggtggaacct gaagtcgagc cagaggtgga accagaggtg
      841 gacccagaag tggatcctga gaacgaccag ggcaccggca gccagaacaa tatcgccgag
      901 tgctacagca gctgcaagag cgtgaccggc gtgcagaccg aggaatgcag ctgtagctgc
      961 tcctgcggcg cgccgtcgac ctccatcacg gcctataaga gtgaggggga gtcagcggag
     1021 ttctccttcc cactcaacct tggagaggaa agcctgcagg gagagttgag atggaaggca
     1081 gagaaggctc cttcttccca gtcctggatc accttctccc taaagaacca aaaggtgtct
     1141 gtgcagaagt ctactagcaa ccccaagttc cagctgtccg aaacgctccc actcaccctt
     1201 cagatacccc aggtctccct tcagtttgct ggttctggca acctgaccct gactctggac
     1261 agagggatac tgtatcagga agtgaacctg gtggtgatga aagtgactca gcccgacagc
     1321 aacactttga cctgtgaggt gatgggaccc acctcaccca agatgagact gatcttgaag
     1381 caggagaatc aggaggccag ggtctccagg caggagaaag tgattcaagt gcaggcccct
     1441 gaagcagggg tgtggcaatg tctactgagt gaaggtgaag aggtcaagat ggactccaag
     1501 atccaggttt tatccaaagg gttgaattcc ggcggcggct ccggaggcgg taccggtggc
     1561 ggcgacctag ccccacagat gcttcgagaa ctccaggaga ctaatgcggc gctgcaagac
     1621 gtgagagagc tcttgcgaca gcaggtcaag gagatcacct tcctgaagaa tacggtgatg
     1681 gaatgtgacg cctgcggtct agacagaaac ctgcctccac tagcacctct gggcccacac
     1741 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1801 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1861 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1921 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1981 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2041 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2101 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2161 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2221 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2281 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2341 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     2401 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     2461 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     2521 cattggggcc ctgactacaa agaccatgac ggtgattata aagatcatga catcgattac
     2581 aaggatgacg atgacaaggg ccctcaccac caccatcacc actaattaat taaggatccc
     2641 ccgacctcga cctctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     2701 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttggt cgagatccct
     2761 cggagatctc tagctagagg atcgatcccc gccccggacg aactaaacct gactacgaca
     2821 tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg gtacaacttg
     2881 ccaactgggc cctgttccac atgtgacacg gggggggacc aaacacaaag gggttctctg
     2941 actgtagttg acatccttat aaatggatgt gcacatttgc caacactgag tggctttcat
     3001 cctggagcag actttgcagt ctgtggactg caacacaaca ttgcctttat gtgtaactct
     3061 tggctgaagc tcttacacca atgctggggg acatgtacct cccaggggcc caggaagact
     3121 acgggaggct acaccaacgt caatcagagg ggcctgtgta gctaccgata agcggaccct
     3181 caagagggca ttagcaatag tgtttataag gcccccttgt taaccctaaa cgggtagcat
     3241 atgcttcccg ggtagtagta tatactatcc agactaaccc taattcaata gcatatgtta
     3301 cccaacggga agcatatgct atcgaattag ggttagtaaa agggtcctaa ggaacagcga
     3361 tatctcccac cccatgagct gtcacggttt tatttacatg gggtcaggat tccacgaggg
     3421 tagtgaacca ttttagtcac aagggcagtg gctgaagatc aaggagcggg cagtgaactc
     3481 tcctgaatct tcgcctgctt cttcattctc cttcgtttag ctaatagaat aactgctgag
     3541 ttgtgaacag taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag
     3601 aataaaattt ggacgggggg ttcagtggtg gcattgtgct atgacaccaa tataaccctc
     3661 acaaacccct tgggcaataa atactagtgt aggaatgaaa cattctgaat atctttaaca
     3721 atagaaatcc atggggtggg gacaagccgt aaagactgga tgtccatctc acacgaattt
     3781 atggctatgg gcaacacata atcctagtgc aatatgatac tggggttatt aagatgtgtc
     3841 ccaggcaggg accaagacag gtgaaccatg ttgttacact ctatttgtaa caaggggaaa
     3901 gagagtggac gccgacagca gcggactcca ctggttgtct ctaacacccc cgaaaattaa
     3961 acggggctcc acgccaatgg ggcccataaa caaagacaag tggccactct tttttttgaa
     4021 attgtggagt gggggcacgc gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa
     4081 ataagggtgt aataacttgg ctgattgtaa ccccgctaac cactgcggtc aaaccacttg
     4141 cccacaaaac cactaatggc accccgggga atacctgcat aagtaggtgg gcgggccaag
     4201 ataggggcgc gattgctgcg atctggagga caaattacac acacttgcgc ctgagcgcca
     4261 agcacagggt tgttggtcct catattcacg aggtcgctga gagcacggtg ggctaatgtt
     4321 gccatgggta gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc
     4381 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     4441 tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc
     4501 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc
     4561 cgggtagcat atgctatcct aatagagatt agggtagtat atgctatcct aatttatatc
     4621 tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg
     4681 tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg
     4741 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg
     4801 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     4861 tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctcatg ataagctgtc
     4921 aaacatgaga attttcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4981 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5041 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5101 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5161 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5221 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5281 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5341 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     5401 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5461 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5521 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5581 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5641 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     5701 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5761 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5821 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5881 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5941 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6001 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6061 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6121 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6181 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6241 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6301 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6361 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6421 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6481 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6541 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6601 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6661 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6721 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6781 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6841 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6901 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6961 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     7021 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     7081 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     7141 ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag gtcgaccaat
     7201 tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat tgctgcaggc
     7261 gcagaactgg taggtatgga agatctatac attgaatcaa tattggcaat tagccatatt
     7321 agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct
     7381 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gttgacattg
     7441 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     7501 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     7561 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     7621 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     7681 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     7741 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
     7801 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
     7861 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     7921 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
     7981 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcct
     8041 cactctcttc cgcatcgctg tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa
     8101 ctcttcgcgg tctttccagt actcttggat cggaaacccg tcggcctccg aacggtactc
     8161 cgccaccgag ggacctgagc gagtccgcat cgaccggatc ggaaaacct
//
