LOCUS       sequence-224711-        8678 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 8678)
  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 8678)
  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_0910300-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8678
                     /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             2212..2997
                     /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             3004..3069
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3076..3093
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3129..3184
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3183..3202)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5420..5438)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5506..5610
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5611..5679,5680..6471)
                     /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      6642..7230
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7131..7150
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7384..7401
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7518..7539
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7554..7559,7560..7577,7578..7584)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7592..7608
                     /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     7597..7619
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7616..7632
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7616..7632
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7903..8282
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8283..8486
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8437..8457
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8483..8507
                     /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 ggccgccgag
      421 gaattcgaga agatcaagta ccctcaggac gtgaccaaga ttaagtacta cgacaacagc
      481 cacaagatcg acagcaagta cctgtacaag cacgtggcca agggcatgct gcaggacctg
      541 atcgtgaccg tgcagatcga caaggacaga gaggtggaca agatcctgtt cctgcccaag
      601 aaaaagacct acatgaagat catcggcgat tggagcgact accccaaaga gaagaacaac
      661 gagatctgcc tggccttcgt gctgcagaag tactttaaga aagagaaaga aaaaaagtgc
      721 accaaggatt tcttcaccag cggcaacatc tacaacgtgt gctacaacga caacaactac
      781 gccatcaaga acctgtacct ctacatcgag atcagccaga agtacctcga gaagctgttc
      841 agcagctaca gcatccccgt gatcgtgaac accgagaaca acatcatcta cttcgccaac
      901 gacaagatga agtccaaaga ggaaaagccc atcctggacc tgcacatctg ctacaagagc
      961 aactacgaga cacccaccta cctgggcaag ctgatcttca actcctaccc catcaacatc
     1021 aaccggaaca aggaccagtt cgacattatc tacaccaaga gctttctgac caacaactgg
     1081 gactacaacc tggccatcaa gggcaactac atcaccagct acaaccgggt ggtgttcgtg
     1141 cggatcccca gagacagccc tcccaacaaa atgaagtgcc cacccttctg ggattacagc
     1201 gtgctgggca gcggccctct gcacaagatc aagaagaaaa acaagaacaa tatcattcag
     1261 aacatggtgc agtacagctt cagcaacaag aatatccaga actactacat cccattcctc
     1321 aagaaggacg agctgcagac caacgaggcc tacatcatct gcctgtacag cgacgagaac
     1381 gacttcgagg gcatgcagat catcaacgcc cacttctaca tcaacaccgc cgacagcggc
     1441 gcgccgtcga cctccatcac ggcctataag agtgaggggg agtcagcgga gttctccttc
     1501 ccactcaacc ttggagagga aagcctgcag ggagagttga gatggaaggc agagaaggct
     1561 ccttcttccc agtcctggat caccttctcc ctaaagaacc aaaaggtgtc tgtgcagaag
     1621 tctactagca accccaagtt ccagctgtcc gaaacgctcc cactcaccct tcagataccc
     1681 caggtctccc ttcagtttgc tggttctggc aacctgaccc tgactctgga cagagggata
     1741 ctgtatcagg aagtgaacct ggtggtgatg aaagtgactc agcccgacag caacactttg
     1801 acctgtgagg tgatgggacc cacctcaccc aagatgagac tgatcttgaa gcaggagaat
     1861 caggaggcca gggtctccag gcaggagaaa gtgattcaag tgcaggcccc tgaagcaggg
     1921 gtgtggcaat gtctactgag tgaaggtgaa gaggtcaaga tggactccaa gatccaggtt
     1981 ttatccaaag ggttgaattc cggcggcggc tccggaggcg gtaccggtgg cggcgaccta
     2041 gccccacaga tgcttcgaga actccaggag actaatgcgg cgctgcaaga cgtgagagag
     2101 ctcttgcgac agcaggtcaa ggagatcacc ttcctgaaga atacggtgat ggaatgtgac
     2161 gcctgcggtc tagacagaaa cctgcctcca ctagcacctc tgggcccaca cccagaaacg
     2221 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2281 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2341 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2401 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2461 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2521 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2581 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2641 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2701 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2761 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2821 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2881 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2941 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggggc
     3001 cctgactaca aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac
     3061 gatgacaagg gccctcacca ccaccatcac cactaataat aaggatcccc cgacctcgac
     3121 ctctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     3181 ctcactcgga aggacatatg ggagggcaaa tcatttggtc gagatccctc ggagatctct
     3241 agctagagga tcgatccccg ccccggacga actaaacctg actacgacat ctctgcccct
     3301 tcttcgcggg gcagtgcatg taatcccttc agttggttgg tacaacttgc caactgggcc
     3361 ctgttccaca tgtgacacgg ggggggacca aacacaaagg ggttctctga ctgtagttga
     3421 catccttata aatggatgtg cacatttgcc aacactgagt ggctttcatc ctggagcaga
     3481 ctttgcagtc tgtggactgc aacacaacat tgcctttatg tgtaactctt ggctgaagct
     3541 cttacaccaa tgctggggga catgtacctc ccaggggccc aggaagacta cgggaggcta
     3601 caccaacgtc aatcagaggg gcctgtgtag ctaccgataa gcggaccctc aagagggcat
     3661 tagcaatagt gtttataagg cccccttgtt aaccctaaac gggtagcata tgcttcccgg
     3721 gtagtagtat atactatcca gactaaccct aattcaatag catatgttac ccaacgggaa
     3781 gcatatgcta tcgaattagg gttagtaaaa gggtcctaag gaacagcgat atctcccacc
     3841 ccatgagctg tcacggtttt atttacatgg ggtcaggatt ccacgagggt agtgaaccat
     3901 tttagtcaca agggcagtgg ctgaagatca aggagcgggc agtgaactct cctgaatctt
     3961 cgcctgcttc ttcattctcc ttcgtttagc taatagaata actgctgagt tgtgaacagt
     4021 aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga ataaaatttg
     4081 gacggggggt tcagtggtgg cattgtgcta tgacaccaat ataaccctca caaacccctt
     4141 gggcaataaa tactagtgta ggaatgaaac attctgaata tctttaacaa tagaaatcca
     4201 tggggtgggg acaagccgta aagactggat gtccatctca cacgaattta tggctatggg
     4261 caacacataa tcctagtgca atatgatact ggggttatta agatgtgtcc caggcaggga
     4321 ccaagacagg tgaaccatgt tgttacactc tatttgtaac aaggggaaag agagtggacg
     4381 ccgacagcag cggactccac tggttgtctc taacaccccc gaaaattaaa cggggctcca
     4441 cgccaatggg gcccataaac aaagacaagt ggccactctt ttttttgaaa ttgtggagtg
     4501 ggggcacgcg tcagccccca cacgccgccc tgcggttttg gactgtaaaa taagggtgta
     4561 ataacttggc tgattgtaac cccgctaacc actgcggtca aaccacttgc ccacaaaacc
     4621 actaatggca ccccggggaa tacctgcata agtaggtggg cgggccaaga taggggcgcg
     4681 attgctgcga tctggaggac aaattacaca cacttgcgcc tgagcgccaa gcacagggtt
     4741 gttggtcctc atattcacga ggtcgctgag agcacggtgg gctaatgttg ccatgggtag
     4801 catatactac ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata
     4861 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     4921 tgctatccta atttatatct gggtagcata ggctatccta atctatatct gggtagcata
     4981 tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata
     5041 tgctatccta atagagatta gggtagtata tgctatccta atttatatct gggtagcata
     5101 tactacccaa atatctggat agcatatgct atcctaatct atatctgggt agcatatgct
     5161 atcctaatct atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     5221 atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt agcataggct
     5281 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     5341 atcctaatct gtatccgggt agcatatgct atcctcatga taagctgtca aacatgagaa
     5401 ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5461 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5521 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5581 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5641 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5701 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5761 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5821 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     5881 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5941 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6001 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6061 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6121 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     6181 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6241 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6301 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6361 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6421 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6481 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6541 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6601 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6661 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6721 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6781 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6841 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6901 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6961 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7021 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7081 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7141 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7201 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7261 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7321 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7381 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7441 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     7501 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     7561 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     7621 aacagctatg accatgatta cgccaagctc tagctagagg tcgaccaatt ctcatgtttg
     7681 acagcttatc atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg cagaactggt
     7741 aggtatggaa gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
     7801 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
     7861 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
     7921 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     7981 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     8041 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     8101 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     8161 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     8221 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     8281 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
     8341 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     8401 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
     8461 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcctc actctcttcc
     8521 gcatcgctgt ctgcgagggc cagctgttgg gctcgcggtt gaggacaaac tcttcgcggt
     8581 ctttccagta ctcttggatc ggaaacccgt cggcctccga acggtactcc gccaccgagg
     8641 gacctgagcg agtccgcatc gaccggatcg gaaaacct
//
