LOCUS       sequence-224480-        8174 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 8174)
  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 8174)
  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: P28-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8174
                     /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             1708..2493
                     /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             2500..2565
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2572..2589
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2625..2680
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2679..2698)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4916..4934)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5002..5106
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5107..5175,5176..5967)
                     /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      6138..6726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6627..6646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6880..6897
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7014..7035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7050..7055,7056..7073,7074..7080)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7088..7104
                     /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     7093..7115
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7112..7128
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7112..7128
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7399..7778
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7779..7982
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7933..7953
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7979..8003
                     /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 ggccgccaga
      421 gtgaccgaga acaccatctg caaatacggc tacctgatcc agatgagcaa ccactacgag
      481 tgcaagtgca tcgagggcta cgtgctgatc aacgaggata cctgcggcaa gaaagtcgtg
      541 tgcgacaagg tggaaaacag cttcaaggcc tgcgacgagt acgcctactg cttcgacctg
      601 ggcaacaaga acaacgagaa gcagatcaag tgcatgtgcc ggaccgagta caccctgaca
      661 gccggcgtgt gcgtgcccaa cgtgtgccgg gacaaagtgt gcggcaaggg caagtgtatc
      721 gtggaccccg ccaacagcct gacccacacc tgtagctgca acatcggcac catcctgaac
      781 cagaacaagc tgtgcgacat ccagggcgac accccttgca gcctgaagtg cgccgagaac
      841 gaagtgtgca ccctggaagg caactactac acctgtaaag aggaccccag cagcaacggc
      901 ggaggcaaca ccgtggacca ggccgacaca agcggcgcgc cgtcgacctc catcacggcc
      961 tataagagtg agggggagtc agcggagttc tccttcccac tcaaccttgg agaggaaagc
     1021 ctgcagggag agttgagatg gaaggcagag aaggctcctt cttcccagtc ctggatcacc
     1081 ttctccctaa agaaccaaaa ggtgtctgtg cagaagtcta ctagcaaccc caagttccag
     1141 ctgtccgaaa cgctcccact cacccttcag ataccccagg tctcccttca gtttgctggt
     1201 tctggcaacc tgaccctgac tctggacaga gggatactgt atcaggaagt gaacctggtg
     1261 gtgatgaaag tgactcagcc cgacagcaac actttgacct gtgaggtgat gggacccacc
     1321 tcacccaaga tgagactgat cttgaagcag gagaatcagg aggccagggt ctccaggcag
     1381 gagaaagtga ttcaagtgca ggcccctgaa gcaggggtgt ggcaatgtct actgagtgaa
     1441 ggtgaagagg tcaagatgga ctccaagatc caggttttat ccaaagggtt gaattccggc
     1501 ggcggctccg gaggcggtac cggtggcggc gacctagccc cacagatgct tcgagaactc
     1561 caggagacta atgcggcgct gcaagacgtg agagagctct tgcgacagca ggtcaaggag
     1621 atcaccttcc tgaagaatac ggtgatggaa tgtgacgcct gcggtctaga cagaaacctg
     1681 cctccactag cacctctggg cccacaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1741 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1801 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1861 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     1921 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1981 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2041 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2101 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2161 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2221 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2281 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2341 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2401 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2461 gctgagatag gtgcctcact gattaagcat tggggccctg actacaaaga ccatgacggt
     2521 gattataaag atcatgacat cgattacaag gatgacgatg acaagggccc tcaccaccac
     2581 catcaccact aataataagg atcccccgac ctcgacctct ggctaataaa ggaaatttat
     2641 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     2701 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc
     2761 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
     2821 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
     2881 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
     2941 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
     3001 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
     3061 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
     3121 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
     3181 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
     3241 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
     3301 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
     3361 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
     3421 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
     3481 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
     3541 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
     3601 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
     3661 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
     3721 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
     3781 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
     3841 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
     3901 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
     3961 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
     4021 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
     4081 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
     4141 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
     4201 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
     4261 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
     4321 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     4381 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     4441 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4501 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
     4561 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
     4621 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
     4681 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4741 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4801 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     4861 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc
     4921 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4981 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5041 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5101 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5161 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5221 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5281 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5341 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5401 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5461 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5521 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5581 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5641 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5701 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5761 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5821 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5881 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5941 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6001 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6061 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6121 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6181 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6241 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6301 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6361 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6421 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6481 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6541 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6601 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6661 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6721 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6781 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6841 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6901 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     6961 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7021 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     7081 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     7141 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg
     7201 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga
     7261 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
     7321 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     7381 caatatgacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     7441 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     7501 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     7561 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     7621 cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg
     7681 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt
     7741 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     7801 ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     7861 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc
     7921 ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     7981 ctcgtttagt gaaccgtcag atcctcactc tcttccgcat cgctgtctgc gagggccagc
     8041 tgttgggctc gcggttgagg acaaactctt cgcggtcttt ccagtactct tggatcggaa
     8101 acccgtcggc ctccgaacgg tactccgcca ccgagggacc tgagcgagtc cgcatcgacc
     8161 ggatcggaaa acct
//
