LOCUS       sequence-224567-        8318 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 8318)
  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 8318)
  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: AOP-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8318
                     /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             1852..2637
                     /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             2644..2709
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2716..2733
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2769..2824
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2823..2842)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5060..5078)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5146..5250
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5251..5319,5320..6111)
                     /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      6282..6870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6771..6790
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7024..7041
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7158..7179
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7194..7199,7200..7217,7218..7224)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7232..7248
                     /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     7237..7259
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7256..7272
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7256..7272
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7543..7922
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7923..8126
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8077..8097
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8123..8147
                     /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 ggccgccttc
      421 aagaacgccc tgatcaagca cagcatcaac atcgtgtcca agcggggcaa cagcaagaac
      481 agattcagcc agaaggtgta cgagtccaag aacatcgacc tcgagaacga catcaaagag
      541 aacgatctga tccccaacgt gaaagtgatg atcgacgtgc ggaacatgaa caatatcgcc
      601 gacaccgacg gcagccccaa cgacttcacc agcatcgaca cccacgagct gttcaacaac
      661 aagaagatcc tgctgatctc cctgcctggc gccttcaccc ctacctgcag caccaagatg
      721 atccccggct acgaggaaga gtacgactac ttcatcaaag aaaacaactt cgacgacatc
      781 tactgcatca ccaacaacga tatctacgtg ctgaagtcct ggttcaagag catggacatc
      841 aagaagatta agtacatcag cgacggcaac agcgccttta ccgagagcat gaacatgctg
      901 gtggacaaga gcaacttctt catgggcatg cggccttgga gattcgtggc catcgtggaa
      961 aacaacatcc tggtcaagat gttccaggaa aaggacaagc agcacaacat ccagaccgac
     1021 ccctacgaca tcagcaccgt gaacaatgtg aaagagttcc tgaagaacaa ccagctcggc
     1081 gcgccgtcga cctccatcac ggcctataag agtgaggggg agtcagcgga gttctccttc
     1141 ccactcaacc ttggagagga aagcctgcag ggagagttga gatggaaggc agagaaggct
     1201 ccttcttccc agtcctggat caccttctcc ctaaagaacc aaaaggtgtc tgtgcagaag
     1261 tctactagca accccaagtt ccagctgtcc gaaacgctcc cactcaccct tcagataccc
     1321 caggtctccc ttcagtttgc tggttctggc aacctgaccc tgactctgga cagagggata
     1381 ctgtatcagg aagtgaacct ggtggtgatg aaagtgactc agcccgacag caacactttg
     1441 acctgtgagg tgatgggacc cacctcaccc aagatgagac tgatcttgaa gcaggagaat
     1501 caggaggcca gggtctccag gcaggagaaa gtgattcaag tgcaggcccc tgaagcaggg
     1561 gtgtggcaat gtctactgag tgaaggtgaa gaggtcaaga tggactccaa gatccaggtt
     1621 ttatccaaag ggttgaattc cggcggcggc tccggaggcg gtaccggtgg cggcgaccta
     1681 gccccacaga tgcttcgaga actccaggag actaatgcgg cgctgcaaga cgtgagagag
     1741 ctcttgcgac agcaggtcaa ggagatcacc ttcctgaaga atacggtgat ggaatgtgac
     1801 gcctgcggtc tagacagaaa cctgcctcca ctagcacctc tgggcccaca cccagaaacg
     1861 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1921 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1981 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2041 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2101 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2161 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2221 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2281 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2341 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2401 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2461 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2521 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2581 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggggc
     2641 cctgactaca aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac
     2701 gatgacaagg gccctcacca ccaccatcac cactaataat aaggatcccc cgacctcgac
     2761 ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     2821 ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc ggagatctct
     2881 agctagagga tcgatccccg ccccggacga actaaacctg actacgacat ctctgcccct
     2941 tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc caactgggcc
     3001 ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga ctgtagttga
     3061 catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc ctggagcaga
     3121 ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt ggctgaagct
     3181 cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta cgggaggcta
     3241 caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc aagagggcat
     3301 tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata tgcttcccgg
     3361 gtagtagtat atactatcca gactaaccct aattcaatag catatgttac ccaacgggaa
     3421 gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat atctcccacc
     3481 ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt agtgaaccat
     3541 tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct cctgaatctt
     3601 cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt tgtgaacagt
     3661 aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga ataaaatttg
     3721 gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca caaacccctt
     3781 gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa tagaaatcca
     3841 tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta tggctatggg
     3901 caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc caggcaggga
     3961 ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag agagtggacg
     4021 ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa cggggctcca
     4081 cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa ttgtggagtg
     4141 ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa taagggtgta
     4201 ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc ccacaaaacc
     4261 actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga taggggcgcg
     4321 attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa gcacagggtt
     4381 gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg ccatgggtag
     4441 catatactac ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata
     4501 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     4561 tgctatccta atttatatct gggtagcata ggctatccta atctatatct gggtagcata
     4621 tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata
     4681 tgctatccta atagagatta gggtagtata tgctatccta atttatatct gggtagcata
     4741 tactacccaa atatctggat agcatatgct atcctaatct atatctgggt agcatatgct
     4801 atcctaatct atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     4861 atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt agcataggct
     4921 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     4981 atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca aacatgagaa
     5041 ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5101 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5161 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5221 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5281 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5341 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5401 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5461 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     5521 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5581 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5641 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5701 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5761 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     5821 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5881 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5941 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6001 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6061 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6121 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6181 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6241 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6301 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6361 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6421 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6481 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6541 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6601 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6661 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6721 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6781 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6841 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6901 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6961 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7021 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7081 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     7141 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     7201 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     7261 aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt ctcatgtttg
     7321 acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg cagaactggt
     7381 aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
     7441 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
     7501 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
     7561 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     7621 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     7681 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     7741 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     7801 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     7861 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     7921 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
     7981 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     8041 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
     8101 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcctc actctcttcc
     8161 gcatcgctgt ctgcgagggc cagctgttgg gctcgcggtt gaggacaaac tcttcgcggt
     8221 ctttccagta ctcttggatc ggaaacccgt cggcctccga acggtactcc gccaccgagg
     8281 gacctgagcg agtccgcatc gaccggatcg gaaaacct
//
