LOCUS       sequence-224708-        9011 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 9011)
  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 9011)
  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: POFUT2-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9011
                     /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             2545..3330
                     /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             3337..3402
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3409..3426
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3462..3517
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3516..3535)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5753..5771)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5839..5943
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5944..6012,6013..6804)
                     /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      6975..7563
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7464..7483
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7717..7734
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7851..7872
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7887..7892,7893..7910,7911..7917)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7925..7941
                     /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     7930..7952
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7949..7965
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7949..7965
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8236..8615
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8616..8819
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8770..8790
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8816..8840
                     /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 ggccgccgtg
      421 atctgcgtga cccctcagaa gctgatctgc ctgaaagaag atgtgtacct gggcgatttc
      481 ttcttcttcc tgaaacggaa aaagtacatt atgtacgacg tgaacatcgg cgagggcttc
      541 aacctgcaga aagagatctt ctaccggctg agcctggtca tctacaacct gaacaagaag
      601 gacaagatca acatctacta cctggtgctg cctccctggt gctacgtgac ccactggaac
      661 atccggaagg gcaacaacct gagatgggag ttcttcttca acaccgacat catgaagaaa
      721 gtgatcccca tcatcgagta cgaggaatac gagaagctgt acggcaacta cgccgatatc
      781 atgatcaaca gcaagtacat cctggacaac tacaaagaga agtccttcct gatcctgccc
      841 ttcgaggaat gcaacatcaa cgtgaacagg ttcaagcagt tctgcaagaa gtgcgagcac
      901 aagtacaacg tgctgtactc cggctactgc accaccatca acaccaagca gagcgagtgc
      961 tacagctaca acatgatcag caactacttc atcacctcta tcctcgagaa cctcttcctg
     1021 tacaatatcg ccagcgtgct gatcaagcag tccaccaaca tcctggtgcc cttcgtgaac
     1081 gagctgtacc agagcaacct ggaagatatc ctgctgttca acaacaagct gctgagctac
     1141 ggcaacaact acatcagcaa catcctcaag accaaccact acatctccag ccacctgaga
     1201 tacaccgact ttaagtacat ctcccggtac aacgtcccac ccatccacat tgccctgctg
     1261 aagctgctgt acatcatgtt catcaacaat tgccggatca tctttatcgc ctccgacgag
     1321 aaggtggaaa tccagaaagt cattaacaag gacttccacc agtacaagaa gcacttctac
     1381 ttctacaaca accagaacaa cctccacgag ggcgagttct ctatcatcga gcagtggatc
     1441 tgcacccggt cctacatctt catcggcaac atcttcagcc ggttcaccat gaacatcaac
     1501 tgggagcggc acctgatcaa caagggccag atcaatcaga acatcgacct gtgcagctac
     1561 cacatcaacg acgacaacga ccaggacatc aagaactcct acaagaagat cgtccacatc
     1621 tttaaccaca aggccctgca gaagatcaag aacatctacg acaattacgc cgaccgggac
     1681 aagaagtaca tcaataccat ctgctacaac tttctgagcc acttccccaa caaccgggcc
     1741 atctacagaa aagagtacat caccaacacc ggcgcgccgt cgacctccat cacggcctat
     1801 aagagtgagg gggagtcagc ggagttctcc ttcccactca accttggaga ggaaagcctg
     1861 cagggagagt tgagatggaa ggcagagaag gctccttctt cccagtcctg gatcaccttc
     1921 tccctaaaga accaaaaggt gtctgtgcag aagtctacta gcaaccccaa gttccagctg
     1981 tccgaaacgc tcccactcac ccttcagata ccccaggtct cccttcagtt tgctggttct
     2041 ggcaacctga ccctgactct ggacagaggg atactgtatc aggaagtgaa cctggtggtg
     2101 atgaaagtga ctcagcccga cagcaacact ttgacctgtg aggtgatggg acccacctca
     2161 cccaagatga gactgatctt gaagcaggag aatcaggagg ccagggtctc caggcaggag
     2221 aaagtgattc aagtgcaggc ccctgaagca ggggtgtggc aatgtctact gagtgaaggt
     2281 gaagaggtca agatggactc caagatccag gttttatcca aagggttgaa ttccggcggc
     2341 ggctccggag gcggtaccgg tggcggcgac ctagccccac agatgcttcg agaactccag
     2401 gagactaatg cggcgctgca agacgtgaga gagctcttgc gacagcaggt caaggagatc
     2461 accttcctga agaatacggt gatggaatgt gacgcctgcg gtctagacag aaacctgcct
     2521 ccactagcac ctctgggccc acacccagaa acgctggtga aagtaaaaga tgctgaagat
     2581 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2641 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2701 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2761 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2821 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2881 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2941 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3001 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3061 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3121 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3181 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3241 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3301 gagataggtg cctcactgat taagcattgg ggccctgact acaaagacca tgacggtgat
     3361 tataaagatc atgacatcga ttacaaggat gacgatgaca agggccctca ccaccaccat
     3421 caccactaat aataaggatc ccccgacctc gacctctggc taataaagga aatttatttt
     3481 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     3541 aaatcatttg gtcgagatcc ctcggagatc tctagctaga ggatcgatcc ccgccccgga
     3601 cgaactaaac ctgactacga catctctgcc ccttcttcgc ggggcagtgc atgtaatccc
     3661 ttcagttggt tggtacaact tgccaactgg gccctgttcc acatgtgaca cgggggggga
     3721 ccaaacacaa aggggttctc tgactgtagt tgacatcctt ataaatggat gtgcacattt
     3781 gccaacactg agtggctttc atcctggagc agactttgca gtctgtggac tgcaacacaa
     3841 cattgccttt atgtgtaact cttggctgaa gctcttacac caatgctggg ggacatgtac
     3901 ctcccagggg cccaggaaga ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg
     3961 tagctaccga taagcggacc ctcaagaggg cattagcaat agtgtttata aggccccctt
     4021 gttaacccta aacgggtagc atatgcttcc cgggtagtag tatatactat ccagactaac
     4081 cctaattcaa tagcatatgt tacccaacgg gaagcatatg ctatcgaatt agggttagta
     4141 aaagggtcct aaggaacagc gatatctccc accccatgag ctgtcacggt tttatttaca
     4201 tggggtcagg attccacgag ggtagtgaac cattttagtc acaagggcag tggctgaaga
     4261 tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc ttcttcattc tccttcgttt
     4321 agctaataga ataactgctg agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca
     4381 aggtttcagg tgacgccccc agaataaaat ttggacgggg ggttcagtgg tggcattgtg
     4441 ctatgacacc aatataaccc tcacaaaccc cttgggcaat aaatactagt gtaggaatga
     4501 aacattctga atatctttaa caatagaaat ccatggggtg gggacaagcc gtaaagactg
     4561 gatgtccatc tcacacgaat ttatggctat gggcaacaca taatcctagt gcaatatgat
     4621 actggggtta ttaagatgtg tcccaggcag ggaccaagac aggtgaacca tgttgttaca
     4681 ctctatttgt aacaagggga aagagagtgg acgccgacag cagcggactc cactggttgt
     4741 ctctaacacc cccgaaaatt aaacggggct ccacgccaat ggggcccata aacaaagaca
     4801 agtggccact cttttttttg aaattgtgga gtgggggcac gcgtcagccc ccacacgccg
     4861 ccctgcggtt ttggactgta aaataagggt gtaataactt ggctgattgt aaccccgcta
     4921 accactgcgg tcaaaccact tgcccacaaa accactaatg gcaccccggg gaatacctgc
     4981 ataagtaggt gggcgggcca agataggggc gcgattgctg cgatctggag gacaaattac
     5041 acacacttgc gcctgagcgc caagcacagg gttgttggtc ctcatattca cgaggtcgct
     5101 gagagcacgg tgggctaatg ttgccatggg tagcatatac tacccaaata tctggatagc
     5161 atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc
     5221 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc
     5281 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     5341 atatgctatc ctaatctgta tccgggtagc atatgctatc ctaatagaga ttagggtagt
     5401 atatgctatc ctaatttata tctgggtagc atatactacc caaatatctg gatagcatat
     5461 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagcatag
     5521 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     5581 gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     5641 gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat
     5701 gctatcctca tgataagctg tcaaacatga gaattttctt gaagacgaaa gggcctcgtg
     5761 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5821 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5881 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5941 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6001 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6061 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6121 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6181 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6241 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6301 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6361 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6421 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6481 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6541 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6601 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6661 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6721 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6781 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6841 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6901 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6961 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7021 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7081 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7141 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7201 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7261 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7321 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7381 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7441 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7501 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7561 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7621 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7681 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7741 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7801 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7861 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7921 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7981 ctctagctag aggtcgacca attctcatgt ttgacagctt atcatcgcag atccgggcaa
     8041 cgttgttgcc attgctgcag gcgcagaact ggtaggtatg gaagatctat acattgaatc
     8101 aatattggca attagccata ttagtcattg gttatatagc ataaatcaat attggctatt
     8161 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
     8221 tatgaccgcc atgttgacat tgattattga ctagttatta atagtaatca attacggggt
     8281 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     8341 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     8401 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     8461 acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg
     8521 gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc
     8581 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca
     8641 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     8701 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg
     8761 ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     8821 gtttagtgaa ccgtcagatc ctcactctct tccgcatcgc tgtctgcgag ggccagctgt
     8881 tgggctcgcg gttgaggaca aactcttcgc ggtctttcca gtactcttgg atcggaaacc
     8941 cgtcggcctc cgaacggtac tccgccaccg agggacctga gcgagtccgc atcgaccgga
     9001 tcggaaaacc t
//
