LOCUS       sequence-224559-        8330 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 8330)
  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 8330)
  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: SPECT1-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8330
                     /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             1864..2649
                     /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             2656..2721
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2728..2745
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2781..2836
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2835..2854)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5072..5090)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5158..5262
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5263..5331,5332..6123)
                     /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      6294..6882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6783..6802
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7036..7053
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7170..7191
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7206..7211,7212..7229,7230..7236)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7244..7260
                     /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     7249..7271
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7268..7284
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7268..7284
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7555..7934
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7935..8138
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8089..8109
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8135..8159
                     /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 ggccgcctac
      421 aacctgaaca acgacctgag caagaacaac aacttcgccc tgaacaccta cgtgcggaag
      481 gacgacgtgg aagatgacag caagaacgag atcgtggaca acatccagaa gatggtcgac
      541 gacttcagcg acgacatcgg cttcgtgaaa accagcatga gagaggtgct gctggacacc
      601 gaggccagcc tggaagaggt gtccgatcac gtggtgcaga atatcgccaa gtacagcctg
      661 accatcgagg aaaagctgaa cctgttcgac ggcctgctgg aagagttcat cgaaaacaac
      721 aagggcctga tcagcaacct ggccaagcgg cagcagaagc tgaagggcga caagatcaag
      781 aaagtgtgcg acctgatcct gaagaagctc aagaagctgg aaaacgtcaa caagctgatc
      841 aagtacaaga tcatcctgaa gtacggcaac aaggacaaca agaaagagat gatccagacc
      901 ctgaagaacg aggaaggcct gagcgacgac tttaagaaca atctggccaa ctacgagaca
      961 gagcagaaca acgatgacat caaagagatc gagctggtca acttcatcag caccaattac
     1021 gacaagttcg tggtcaacct cgaggacctc aacaaagagc tgctgaagga cctgaacatg
     1081 gccctgagcg gcgcgccgtc gacctccatc acggcctata agagtgaggg ggagtcagcg
     1141 gagttctcct tcccactcaa ccttggagag gaaagcctgc agggagagtt gagatggaag
     1201 gcagagaagg ctccttcttc ccagtcctgg atcaccttct ccctaaagaa ccaaaaggtg
     1261 tctgtgcaga agtctactag caaccccaag ttccagctgt ccgaaacgct cccactcacc
     1321 cttcagatac cccaggtctc ccttcagttt gctggttctg gcaacctgac cctgactctg
     1381 gacagaggga tactgtatca ggaagtgaac ctggtggtga tgaaagtgac tcagcccgac
     1441 agcaacactt tgacctgtga ggtgatggga cccacctcac ccaagatgag actgatcttg
     1501 aagcaggaga atcaggaggc cagggtctcc aggcaggaga aagtgattca agtgcaggcc
     1561 cctgaagcag gggtgtggca atgtctactg agtgaaggtg aagaggtcaa gatggactcc
     1621 aagatccagg ttttatccaa agggttgaat tccggcggcg gctccggagg cggtaccggt
     1681 ggcggcgacc tagccccaca gatgcttcga gaactccagg agactaatgc ggcgctgcaa
     1741 gacgtgagag agctcttgcg acagcaggtc aaggagatca ccttcctgaa gaatacggtg
     1801 atggaatgtg acgcctgcgg tctagacaga aacctgcctc cactagcacc tctgggccca
     1861 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1921 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1981 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2041 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2101 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2161 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2221 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2281 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2341 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2401 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2461 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2521 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2581 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2641 aagcattggg gccctgacta caaagaccat gacggtgatt ataaagatca tgacatcgat
     2701 tacaaggatg acgatgacaa gggccctcac caccaccatc accactaata ataaggatcc
     2761 cccgacctcg acctctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     2821 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttgg tcgagatccc
     2881 tcggagatct ctagctagag gatcgatccc cgccccggac gaactaaacc tgactacgac
     2941 atctctgccc cttcttcgcg gggcagtgca tgtaatccct tcagttggtt ggtacaactt
     3001 gccaactggg ccctgttcca catgtgacac ggggggggac caaacacaaa ggggttctct
     3061 gactgtagtt gacatcctta taaatggatg tgcacatttg ccaacactga gtggctttca
     3121 tcctggagca gactttgcag tctgtggact gcaacacaac attgccttta tgtgtaactc
     3181 ttggctgaag ctcttacacc aatgctgggg gacatgtacc tcccaggggc ccaggaagac
     3241 tacgggaggc tacaccaacg tcaatcagag gggcctgtgt agctaccgat aagcggaccc
     3301 tcaagagggc attagcaata gtgtttataa ggcccccttg ttaaccctaa acgggtagca
     3361 tatgcttccc gggtagtagt atatactatc cagactaacc ctaattcaat agcatatgtt
     3421 acccaacggg aagcatatgc tatcgaatta gggttagtaa aagggtccta aggaacagcg
     3481 atatctccca ccccatgagc tgtcacggtt ttatttacat ggggtcagga ttccacgagg
     3541 gtagtgaacc attttagtca caagggcagt ggctgaagat caaggagcgg gcagtgaact
     3601 ctcctgaatc ttcgcctgct tcttcattct ccttcgttta gctaatagaa taactgctga
     3661 gttgtgaaca gtaaggtgta tgtgaggtgc tcgaaaacaa ggtttcaggt gacgccccca
     3721 gaataaaatt tggacggggg gttcagtggt ggcattgtgc tatgacacca atataaccct
     3781 cacaaacccc ttgggcaata aatactagtg taggaatgaa acattctgaa tatctttaac
     3841 aatagaaatc catggggtgg ggacaagccg taaagactgg atgtccatct cacacgaatt
     3901 tatggctatg ggcaacacat aatcctagtg caatatgata ctggggttat taagatgtgt
     3961 cccaggcagg gaccaagaca ggtgaaccat gttgttacac tctatttgta acaaggggaa
     4021 agagagtgga cgccgacagc agcggactcc actggttgtc tctaacaccc ccgaaaatta
     4081 aacggggctc cacgccaatg gggcccataa acaaagacaa gtggccactc ttttttttga
     4141 aattgtggag tgggggcacg cgtcagcccc cacacgccgc cctgcggttt tggactgtaa
     4201 aataagggtg taataacttg gctgattgta accccgctaa ccactgcggt caaaccactt
     4261 gcccacaaaa ccactaatgg caccccgggg aatacctgca taagtaggtg ggcgggccaa
     4321 gataggggcg cgattgctgc gatctggagg acaaattaca cacacttgcg cctgagcgcc
     4381 aagcacaggg ttgttggtcc tcatattcac gaggtcgctg agagcacggt gggctaatgt
     4441 tgccatgggt agcatatact acccaaatat ctggatagca tatgctatcc taatctatat
     4501 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4561 ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat
     4621 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat
     4681 ccgggtagca tatgctatcc taatagagat tagggtagta tatgctatcc taatttatat
     4741 ctgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg
     4801 gtagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg
     4861 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg
     4921 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4981 gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctcat gataagctgt
     5041 caaacatgag aattttcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5101 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5161 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5221 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5281 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5341 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5401 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5461 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     5521 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5581 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5641 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5701 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5761 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     5821 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5881 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5941 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6001 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6061 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6121 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6181 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6241 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6301 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6361 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6421 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6481 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6541 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6601 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6661 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6721 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6781 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6841 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6901 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6961 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     7021 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     7081 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     7141 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     7201 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     7261 tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgaccaa
     7321 ttctcatgtt tgacagctta tcatcgcaga tccgggcaac gttgttgcca ttgctgcagg
     7381 cgcagaactg gtaggtatgg aagatctata cattgaatca atattggcaa ttagccatat
     7441 tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc
     7501 tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca tgttgacatt
     7561 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     7621 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     7681 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     7741 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     7801 atcatatgcc aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     7861 atgcccagta catgacctta cgggactttc ctacttggca gtacatctac gtattagtca
     7921 tcgctattac catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg
     7981 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     8041 aaaatcaacg ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg
     8101 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcc
     8161 tcactctctt ccgcatcgct gtctgcgagg gccagctgtt gggctcgcgg ttgaggacaa
     8221 actcttcgcg gtctttccag tactcttgga tcggaaaccc gtcggcctcc gaacggtact
     8281 ccgccaccga gggacctgag cgagtccgca tcgaccggat cggaaaacct
//
