LOCUS       sequence-224243-        8012 bp    DNA     circular SYN 15-APR-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 8012)
  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 8012)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_1310500-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8012
                     /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             1546..2331
                     /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             2338..2403
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2410..2427
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2463..2518
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2517..2536)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4754..4772)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4840..4944
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4945..5013,5014..5805)
                     /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      5976..6564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6465..6484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6718..6735
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6852..6873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6888..6893,6894..6911,6912..6918)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6926..6942
                     /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     6931..6953
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6950..6966
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6950..6966
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7237..7616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7617..7820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7771..7791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7817..7841
                     /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 gaggacgtga tcgagctgaa gaacaactac atcgagggca tcgtgaaggg cgacgccgac
      481 ctgctgttca actgcgccat ctacctggac agcgacacct acgtgtcccc aaagagcgac
      541 ggcaagttcc tgttcaagaa cgtgaaagag ggcgagtaca ctatgtacgt gaaccacccc
      601 tacctcgagt tcaacaagta ctacgtggaa gtgaagaagc agatcacccg gaacaacggc
      661 gccctgtacc tggtgcaggc ctacgagctg aacctgccct tcgagaagtc caacctgatc
      721 gtgaacaaca tcgtgttcaa ggccaagcgg atctacgact tcctgatccc cggcgcgccg
      781 tcgacctcca tcacggccta taagagtgag ggggagtcag cggagttctc cttcccactc
      841 aaccttggag aggaaagcct gcagggagag ttgagatgga aggcagagaa ggctccttct
      901 tcccagtcct ggatcacctt ctccctaaag aaccaaaagg tgtctgtgca gaagtctact
      961 agcaacccca agttccagct gtccgaaacg ctcccactca cccttcagat accccaggtc
     1021 tcccttcagt ttgctggttc tggcaacctg accctgactc tggacagagg gatactgtat
     1081 caggaagtga acctggtggt gatgaaagtg actcagcccg acagcaacac tttgacctgt
     1141 gaggtgatgg gacccacctc acccaagatg agactgatct tgaagcagga gaatcaggag
     1201 gccagggtct ccaggcagga gaaagtgatt caagtgcagg cccctgaagc aggggtgtgg
     1261 caatgtctac tgagtgaagg tgaagaggtc aagatggact ccaagatcca ggttttatcc
     1321 aaagggttga attccggcgg cggctccgga ggcggtaccg gtggcggcga cctagcccca
     1381 cagatgcttc gagaactcca ggagactaat gcggcgctgc aagacgtgag agagctcttg
     1441 cgacagcagg tcaaggagat caccttcctg aagaatacgg tgatggaatg tgacgcctgc
     1501 ggtctagaca gaaacctgcc tccactagca cctctgggcc cacacccaga aacgctggtg
     1561 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1621 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1681 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     1741 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1801 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     1861 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     1921 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     1981 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2041 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2101 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2161 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2221 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2281 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gggccctgac
     2341 tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac
     2401 aagggccctc accaccacca tcaccactaa taataaggat cccccgacct cgacctctgg
     2461 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     2521 cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag
     2581 aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg
     2641 cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc
     2701 cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct
     2761 tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc
     2821 agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca
     2881 ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa
     2941 cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa
     3001 tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta
     3061 gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat
     3121 gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga
     3181 gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt
     3241 cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg
     3301 cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg
     3361 tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg
     3421 gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa
     3481 taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt
     3541 ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac
     3601 ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga
     3661 caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca
     3721 gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa
     3781 tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca
     3841 cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact
     3901 tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat
     3961 ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct
     4021 gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt
     4081 cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata
     4141 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag cataggctat
     4201 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4261 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4321 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     4381 cctaatagag attagggtag tatatgctat cctaatttat atctgggtag catatactac
     4441 ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata tgctatccta
     4501 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     4561 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     4621 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     4681 atctgtatcc gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct
     4741 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4801 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4861 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4921 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4981 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5041 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5101 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5161 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     5221 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5281 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5341 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5401 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5461 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     5521 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5581 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5641 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5701 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5761 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5821 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5881 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5941 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6001 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6061 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6121 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6181 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6241 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6301 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6361 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6421 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6481 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6541 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6601 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6661 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6721 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     6781 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     6841 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     6901 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     6961 tatgaccatg attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct
     7021 tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat
     7081 ggaagatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     7141 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     7201 tttatattgg ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt
     7261 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     7321 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     7381 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     7441 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc
     7501 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     7561 tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     7621 tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     7681 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     7741 caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg
     7801 aggtctatat aagcagagct cgtttagtga accgtcagat cctcactctc ttccgcatcg
     7861 ctgtctgcga gggccagctg ttgggctcgc ggttgaggac aaactcttcg cggtctttcc
     7921 agtactcttg gatcggaaac ccgtcggcct ccgaacggta ctccgccacc gagggacctg
     7981 agcgagtccg catcgaccgg atcggaaaac ct
//
