LOCUS       sequence-224488-        8207 bp    DNA     circular SYN 14-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 8207)
  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 8207)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: SOAP-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8207
                     /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             1741..2526
                     /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             2533..2598
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2605..2622
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2658..2713
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2712..2731)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4949..4967)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5035..5139
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5140..5208,5209..6000)
                     /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      6171..6759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6660..6679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6913..6930
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7047..7068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7083..7088,7089..7106,7107..7113)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7121..7137
                     /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     7126..7148
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7145..7161
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7145..7161
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7432..7811
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7812..8015
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7966..7986
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8012..8036
                     /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 ggccgccaaa
      421 ggcgtgcgga acaaggccga gaacagagcc ccttacgtgc actaccacaa caacgccaac
      481 atcaagaact acgccgccgt ggacatcttc agccccaaga aaaccggcaa agaatgcatc
      541 aagtgcctgc ccgacaactt ctgcgagtgc gagtgctcct gcaagaacaa ggctggcttc
      601 agcatgaagt accggcacgc cagcaagggc tccaagggct acagcaagaa gatgaccaag
      661 accagcagcc ggtcccggca gtgcaacgac ctgcccctgg aaaagagcga gcaggaaacc
      721 gagcccgagc tggaacctga ggtggaaccc gaggtcgagc ctgaagtcga acccgaagtc
      781 gaaccagagg tcgagcccga ggtggaacct gaagtcgagc cagaggtgga accagaggtg
      841 gacccagaag tggatcctga gaacgaccag ggcaccggca gccagaacaa tatcgccgag
      901 tgctacagca gctgcaagag cgtgaccggc gtgcagaccg aggaatgcag ctgtagctgc
      961 tcctgcggcg cgccgtcgac ctccatcacg gcctataaga gtgaggggga gtcagcggag
     1021 ttctccttcc cactcaacct tggagaggaa agcctgcagg gagagttgag atggaaggca
     1081 gagaaggctc cttcttccca gtcctggatc accttctccc taaagaacca aaaggtgtct
     1141 gtgcagaagt ctactagcaa ccccaagttc cagctgtccg aaacgctccc actcaccctt
     1201 cagatacccc aggtctccct tcagtttgct ggttctggca acctgaccct gactctggac
     1261 agagggatac tgtatcagga agtgaacctg gtggtgatga aagtgactca gcccgacagc
     1321 aacactttga cctgtgaggt gatgggaccc acctcaccca agatgagact gatcttgaag
     1381 caggagaatc aggaggccag ggtctccagg caggagaaag tgattcaagt gcaggcccct
     1441 gaagcagggg tgtggcaatg tctactgagt gaaggtgaag aggtcaagat ggactccaag
     1501 atccaggttt tatccaaagg gttgaattcc ggcggcggct ccggaggcgg taccggtggc
     1561 ggcgacctag ccccacagat gcttcgagaa ctccaggaga ctaatgcggc gctgcaagac
     1621 gtgagagagc tcttgcgaca gcaggtcaag gagatcacct tcctgaagaa tacggtgatg
     1681 gaatgtgacg cctgcggtct agacagaaac ctgcctccac tagcacctct gggcccacac
     1741 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1801 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1861 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1921 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1981 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2041 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2101 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2161 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2221 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2281 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2341 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     2401 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     2461 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     2521 cattggggcc ctgactacaa agaccatgac ggtgattata aagatcatga catcgattac
     2581 aaggatgacg atgacaaggg ccctcaccac caccatcacc actaataata aggatccccc
     2641 gacctcgacc tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     2701 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttggtcg agatccctcg
     2761 gagatctcta gctagaggat cgatccccgc cccggacgaa ctaaacctga ctacgacatc
     2821 tctgcccctt cttcgcgggg cagtgcatgt aatcccttca gttggttggt acaacttgcc
     2881 aactgggccc tgttccacat gtgacacggg gggggaccaa acacaaaggg gttctctgac
     2941 tgtagttgac atccttataa atggatgtgc acatttgcca acactgagtg gctttcatcc
     3001 tggagcagac tttgcagtct gtggactgca acacaacatt gcctttatgt gtaactcttg
     3061 gctgaagctc ttacaccaat gctgggggac atgtacctcc caggggccca ggaagactac
     3121 gggaggctac accaacgtca atcagagggg cctgtgtagc taccgataag cggaccctca
     3181 agagggcatt agcaatagtg tttataaggc ccccttgtta accctaaacg ggtagcatat
     3241 gcttcccggg tagtagtata tactatccag actaacccta attcaatagc atatgttacc
     3301 caacgggaag catatgctat cgaattaggg ttagtaaaag ggtcctaagg aacagcgata
     3361 tctcccaccc catgagctgt cacggtttta tttacatggg gtcaggattc cacgagggta
     3421 gtgaaccatt ttagtcacaa gggcagtggc tgaagatcaa ggagcgggca gtgaactctc
     3481 ctgaatcttc gcctgcttct tcattctcct tcgtttagct aatagaataa ctgctgagtt
     3541 gtgaacagta aggtgtatgt gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa
     3601 taaaatttgg acggggggtt cagtggtggc attgtgctat gacaccaata taaccctcac
     3661 aaaccccttg ggcaataaat actagtgtag gaatgaaaca ttctgaatat ctttaacaat
     3721 agaaatccat ggggtgggga caagccgtaa agactggatg tccatctcac acgaatttat
     3781 ggctatgggc aacacataat cctagtgcaa tatgatactg gggttattaa gatgtgtccc
     3841 aggcagggac caagacaggt gaaccatgtt gttacactct atttgtaaca aggggaaaga
     3901 gagtggacgc cgacagcagc ggactccact ggttgtctct aacacccccg aaaattaaac
     3961 ggggctccac gccaatgggg cccataaaca aagacaagtg gccactcttt tttttgaaat
     4021 tgtggagtgg gggcacgcgt cagcccccac acgccgccct gcggttttgg actgtaaaat
     4081 aagggtgtaa taacttggct gattgtaacc ccgctaacca ctgcggtcaa accacttgcc
     4141 cacaaaacca ctaatggcac cccggggaat acctgcataa gtaggtgggc gggccaagat
     4201 aggggcgcga ttgctgcgat ctggaggaca aattacacac acttgcgcct gagcgccaag
     4261 cacagggttg ttggtcctca tattcacgag gtcgctgaga gcacggtggg ctaatgttgc
     4321 catgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
     4381 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     4441 ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg
     4501 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg
     4561 ggtagcatat gctatcctaa tagagattag ggtagtatat gctatcctaa tttatatctg
     4621 ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta
     4681 gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta
     4741 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta
     4801 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     4861 gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctcatgat aagctgtcaa
     4921 acatgagaat tttcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4981 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5041 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5101 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5161 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5221 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5281 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5341 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     5401 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5461 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5521 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5581 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5641 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     5701 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5761 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5821 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5881 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5941 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6001 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6061 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6121 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6181 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6241 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6301 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6361 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6421 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6481 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6541 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6601 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6661 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6721 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6781 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6841 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6901 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6961 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     7021 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7081 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7141 cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt cgaccaattc
     7201 tcatgtttga cagcttatca tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc
     7261 agaactggta ggtatggaag atctatacat tgaatcaata ttggcaatta gccatattag
     7321 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
     7381 atcataatat gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat
     7441 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     7501 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     7561 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     7621 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     7681 atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     7741 cccagtacat gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg
     7801 ctattaccat ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact
     7861 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     7921 atcaacggga ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta
     7981 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcctca
     8041 ctctcttccg catcgctgtc tgcgagggcc agctgttggg ctcgcggttg aggacaaact
     8101 cttcgcggtc tttccagtac tcttggatcg gaaacccgtc ggcctccgaa cggtactccg
     8161 ccaccgaggg acctgagcga gtccgcatcg accggatcgg aaaacct
//
