LOCUS       sequence-208766-        8182 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 8182)
  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 8182)
  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: CYP19B-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8182
                     /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             1714..2499
                     /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             2506..2571
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2578..2595
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2633..2688
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2687..2706)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4924..4942)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5010..5114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5115..5183,5184..5975)
                     /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      6146..6734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6635..6654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6888..6905
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7022..7043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7058..7063,7064..7081,7082..7088)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7096..7112
                     /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     7101..7123
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7120..7136
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7120..7136
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7407..7786
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7787..7990
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7941..7961
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7987..8011
                     /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 ggccgccgcc
      421 gaggaacacg agatcaccca caagacctac ttcgacatca ccatcgacga caagcccctg
      481 ggccggatcg tgttcggcct gtacggcaag gtggccccta agaccgtgga aaacttcgtg
      541 tccatctgca agggcaccgt ggtggacggc aagatgctgc actacaccaa cagcatcttc
      601 caccggatca tccccaactt catggcccag ggcggcgata tcaccaactt caacggcgct
      661 ggcggcctga gcatctacgg caagaagttc gaggacgaga acttcaaagt gaaccacgcc
      721 aagcggggcc tgctgagcat ggccaacgcc ggcaagaaca ccaacggcgc ccagttcttc
      781 atcctgttca tccccacccc ttggctggac ggccggcacg tggtgttcgg cgaggtggtg
      841 gaaggcctgg acaagctggt gcacatcgag gccgtgggca ccgacagcgg cgagcctctg
      901 aagcgggtgc tggtcaaaga gtccggcgag ctgcctctcg gcgcgccgtc gacctccatc
      961 acggcctata agagtgaggg ggagtcagcg gagttctcct tcccactcaa ccttggagag
     1021 gaaagcctgc agggagagtt gagatggaag gcagagaagg ctccttcttc ccagtcctgg
     1081 atcaccttct ccctaaagaa ccaaaaggtg tctgtgcaga agtctactag caaccccaag
     1141 ttccagctgt ccgaaacgct cccactcacc cttcagatac cccaggtctc ccttcagttt
     1201 gctggttctg gcaacctgac cctgactctg gacagaggga tactgtatca ggaagtgaac
     1261 ctggtggtga tgaaagtgac tcagcccgac agcaacactt tgacctgtga ggtgatggga
     1321 cccacctcac ccaagatgag actgatcttg aagcaggaga atcaggaggc cagggtctcc
     1381 aggcaggaga aagtgattca agtgcaggcc cctgaagcag gggtgtggca atgtctactg
     1441 agtgaaggtg aagaggtcaa gatggactcc aagatccagg ttttatccaa agggttgaat
     1501 tccggcggcg gctccggagg cggtaccggt ggcggcgacc tagccccaca gatgcttcga
     1561 gaactccagg agactaatgc ggcgctgcaa gacgtgagag agctcttgcg acagcaggtc
     1621 aaggagatca ccttcctgaa gaatacggtg atggaatgtg acgcctgcgg tctagacaga
     1681 aacctgcctc cactagcacc tctgggccca cacccagaaa cgctggtgaa agtaaaagat
     1741 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1801 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1861 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1921 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1981 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2041 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2101 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2161 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     2221 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2281 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2341 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2401 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2461 cagatcgctg agataggtgc ctcactgatt aagcattggg gccctgacta caaagaccat
     2521 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gggccctcac
     2581 caccaccatc accactaatt aattaaggat cccccgacct cgacctctgg ctaataaagg
     2641 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     2701 tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag aggatcgatc
     2761 cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg cggggcagtg
     2821 catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc cacatgtgac
     2881 acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct tataaatgga
     2941 tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc agtctgtgga
     3001 ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca ccaatgctgg
     3061 gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa cgtcaatcag
     3121 aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa tagtgtttat
     3181 aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta gtatatacta
     3241 tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat gctatcgaat
     3301 tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga gctgtcacgg
     3361 ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt cacaagggca
     3421 gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt
     3481 ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg tatgtgaggt
     3541 gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg gggttcagtg
     3601 gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa taaatactag
     3661 tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt ggggacaagc
     3721 cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac ataatcctag
     3781 tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga caggtgaacc
     3841 atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca gcagcggact
     3901 ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa tggggcccat
     3961 aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca cgcgtcagcc
     4021 cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact tggctgattg
     4081 taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg
     4141 ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct gcgatctgga
     4201 ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt cctcatattc
     4261 acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata ctacccaaat
     4321 atctggatag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
     4381 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
     4441 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     4501 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctaatagag
     4561 attagggtag tatatgctat cctaatttat atctgggtag catatactac ccaaatatct
     4621 ggatagcata tgctatccta atctatatct gggtagcata tgctatccta atctatatct
     4681 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     4741 gggtagtata tgctatccta atttatatct gggtagcata ggctatccta atctatatct
     4801 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc
     4861 gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct tgaagacgaa
     4921 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4981 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5041 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5101 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5161 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5221 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5281 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5341 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5401 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5461 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5521 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5581 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5641 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     5701 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5761 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5821 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5881 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5941 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6001 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6061 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6121 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6181 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6241 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6301 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6361 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6421 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6481 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6541 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6601 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6661 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6721 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6781 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6841 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6901 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6961 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     7021 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     7081 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     7141 attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct tatcatcgca
     7201 gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat ggaagatcta
     7261 tacattgaat caatattggc aattagccat attagtcatt ggttatatag cataaatcaa
     7321 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     7381 ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt aatagtaatc
     7441 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     7501 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     7561 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     7621 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga
     7681 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt
     7741 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     7801 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     7861 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     7921 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     7981 aagcagagct cgtttagtga accgtcagat cctcactctc ttccgcatcg ctgtctgcga
     8041 gggccagctg ttgggctcgc ggttgaggac aaactcttcg cggtctttcc agtactcttg
     8101 gatcggaaac ccgtcggcct ccgaacggta ctccgccacc gagggacctg agcgagtccg
     8161 catcgaccgg atcggaaaac ct
//
