LOCUS       sequence-224238-        8807 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 8807)
  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 8807)
  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_0912400-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8807
                     /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             2341..3126
                     /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             3133..3198
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3205..3222
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3258..3313
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3312..3331)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5549..5567)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5635..5739
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5740..5808,5809..6600)
                     /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      6771..7359
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7260..7279
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7513..7530
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7647..7668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7683..7688,7689..7706,7707..7713)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7721..7737
                     /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     7726..7748
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7745..7761
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7745..7761
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8032..8411
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8412..8615
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8566..8586
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8612..8636
                     /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 ggccgcccag
      421 ggcgaccagg tcatcaacga gaagctgacc aacttcgtgt tcctgagctg caactaccag
      481 aagggcaaag tcaacaacaa cctgctgaac agcatcgaga agcggaagcc ccagctgatg
      541 ctgtggatcg gcgactactt ctacaccgag tgcagcgaga tcaagtgcct ggacgacgcc
      601 tacacctaca tcaagaaaga ccccttctac atgaagctga agaagaagtt caagatcgac
      661 ggcatctacg acgaccacga ctacaacaag aacaacggcg accggctgta caagtacaag
      721 aaagagtcca agaagaaata cctggactac ctgaacgtgg acaagaacga cgtgcggtac
      781 aagcggaacg gcgcctacat cagcaagctg tacatcgacc ccgacaatga gaagaaccag
      841 gtcaaaatca tcatgctgga cacccggtac aacaaggacc cctacccttt ctacgcccct
      901 gacagctacc gggacctgtt cgtgcacatg ttcatcagct ttctgagccg gttccacagc
      961 agcatcttcg gcctgtgctg caacagcaag aacgatatcc tgggcaacga gcagtggaag
     1021 tggctggaaa gagagctgac aaacagcaac gccagagccc acatcatcat cagctccacc
     1081 cagatcttca gcaaccacat cattaacgag aactggggcc tgatgcccta cagcctgcgg
     1141 cggctgcgcg agctgatcaa aaagaccaag cccaagggcc tgctgttcct gtccggcgac
     1201 gtgcacttcg gctccatcat cggcaaagag gaatccgtca tcgaagtgac cagcagcagc
     1261 gtgaaccagg aaaacatctt cagctacatc aacaaatacg tgatcttctt cctgaccaac
     1321 atcctgagca aggtgtcccc attcgagctg aacaagatct acagcttcaa caatttcggc
     1381 tccgtgaata tcgcctacgt gaacgacaac gagattaaga tcaagacctc cgtgaacgat
     1441 gccgacggcg tggaaatcct ggtggccaat caggtgttca acaataagaa caatatctat
     1501 accaagacca aggatctgca catcatcctg gacgagttcg ccaccctgga atgcaagagc
     1561 aagaccggcg cgccgtcgac ctccatcacg gcctataaga gtgaggggga gtcagcggag
     1621 ttctccttcc cactcaacct tggagaggaa agcctgcagg gagagttgag atggaaggca
     1681 gagaaggctc cttcttccca gtcctggatc accttctccc taaagaacca aaaggtgtct
     1741 gtgcagaagt ctactagcaa ccccaagttc cagctgtccg aaacgctccc actcaccctt
     1801 cagatacccc aggtctccct tcagtttgct ggttctggca acctgaccct gactctggac
     1861 agagggatac tgtatcagga agtgaacctg gtggtgatga aagtgactca gcccgacagc
     1921 aacactttga cctgtgaggt gatgggaccc acctcaccca agatgagact gatcttgaag
     1981 caggagaatc aggaggccag ggtctccagg caggagaaag tgattcaagt gcaggcccct
     2041 gaagcagggg tgtggcaatg tctactgagt gaaggtgaag aggtcaagat ggactccaag
     2101 atccaggttt tatccaaagg gttgaattcc ggcggcggct ccggaggcgg taccggtggc
     2161 ggcgacctag ccccacagat gcttcgagaa ctccaggaga ctaatgcggc gctgcaagac
     2221 gtgagagagc tcttgcgaca gcaggtcaag gagatcacct tcctgaagaa tacggtgatg
     2281 gaatgtgacg cctgcggtct agacagaaac ctgcctccac tagcacctct gggcccacac
     2341 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2401 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2461 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2521 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2581 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2641 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2701 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2761 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2821 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2881 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2941 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3001 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3061 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3121 cattggggcc ctgactacaa agaccatgac ggtgattata aagatcatga catcgattac
     3181 aaggatgacg atgacaaggg ccctcaccac caccatcacc actaataata aggatccccc
     3241 gacctcgacc tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     3301 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttggtcg agatccctcg
     3361 gagatctcta gctagaggat cgatccccgc cccggacgaa ctaaacctga ctacgacatc
     3421 tctgcccctt cttcgcgggg cagtgcatgt aatcccttca gttggttggt acaacttgcc
     3481 aactgggccc tgttccacat gtgacacggg gggggaccaa acacaaaggg gttctctgac
     3541 tgtagttgac atccttataa atggatgtgc acatttgcca acactgagtg gctttcatcc
     3601 tggagcagac tttgcagtct gtggactgca acacaacatt gcctttatgt gtaactcttg
     3661 gctgaagctc ttacaccaat gctgggggac atgtacctcc caggggccca ggaagactac
     3721 gggaggctac accaacgtca atcagagggg cctgtgtagc taccgataag cggaccctca
     3781 agagggcatt agcaatagtg tttataaggc ccccttgtta accctaaacg ggtagcatat
     3841 gcttcccggg tagtagtata tactatccag actaacccta attcaatagc atatgttacc
     3901 caacgggaag catatgctat cgaattaggg ttagtaaaag ggtcctaagg aacagcgata
     3961 tctcccaccc catgagctgt cacggtttta tttacatggg gtcaggattc cacgagggta
     4021 gtgaaccatt ttagtcacaa gggcagtggc tgaagatcaa ggagcgggca gtgaactctc
     4081 ctgaatcttc gcctgcttct tcattctcct tcgtttagct aatagaataa ctgctgagtt
     4141 gtgaacagta aggtgtatgt gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa
     4201 taaaatttgg acggggggtt cagtggtggc attgtgctat gacaccaata taaccctcac
     4261 aaaccccttg ggcaataaat actagtgtag gaatgaaaca ttctgaatat ctttaacaat
     4321 agaaatccat ggggtgggga caagccgtaa agactggatg tccatctcac acgaatttat
     4381 ggctatgggc aacacataat cctagtgcaa tatgatactg gggttattaa gatgtgtccc
     4441 aggcagggac caagacaggt gaaccatgtt gttacactct atttgtaaca aggggaaaga
     4501 gagtggacgc cgacagcagc ggactccact ggttgtctct aacacccccg aaaattaaac
     4561 ggggctccac gccaatgggg cccataaaca aagacaagtg gccactcttt tttttgaaat
     4621 tgtggagtgg gggcacgcgt cagcccccac acgccgccct gcggttttgg actgtaaaat
     4681 aagggtgtaa taacttggct gattgtaacc ccgctaacca ctgcggtcaa accacttgcc
     4741 cacaaaacca ctaatggcac cccggggaat acctgcataa gtaggtgggc gggccaagat
     4801 aggggcgcga ttgctgcgat ctggaggaca aattacacac acttgcgcct gagcgccaag
     4861 cacagggttg ttggtcctca tattcacgag gtcgctgaga gcacggtggg ctaatgttgc
     4921 catgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
     4981 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     5041 ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg
     5101 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg
     5161 ggtagcatat gctatcctaa tagagattag ggtagtatat gctatcctaa tttatatctg
     5221 ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta
     5281 gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta
     5341 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta
     5401 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     5461 gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctcatgat aagctgtcaa
     5521 acatgagaat tttcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5581 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5641 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5701 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5761 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5821 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5881 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5941 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     6001 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6061 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6121 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6181 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6241 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     6301 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6361 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6421 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6481 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6541 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6601 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6661 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6721 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6781 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6841 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6901 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6961 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7021 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7081 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7141 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7201 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7261 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7321 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7381 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7441 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7501 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     7561 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     7621 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7681 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7741 cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt cgaccaattc
     7801 tcatgtttga cagcttatca tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc
     7861 agaactggta ggtatggaag atctatacat tgaatcaata ttggcaatta gccatattag
     7921 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
     7981 atcataatat gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat
     8041 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     8101 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     8161 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     8221 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     8281 atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     8341 cccagtacat gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg
     8401 ctattaccat ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact
     8461 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     8521 atcaacggga ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta
     8581 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcctca
     8641 ctctcttccg catcgctgtc tgcgagggcc agctgttggg ctcgcggttg aggacaaact
     8701 cttcgcggtc tttccagtac tcttggatcg gaaacccgtc ggcctccgaa cggtactccg
     8761 ccaccgaggg acctgagcga gtccgcatcg accggatcgg aaaacct
//
