LOCUS       sequence-208752-        9448 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 9448)
  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 9448)
  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: PSOP7-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9448
                     /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             2980..3765
                     /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             3772..3837
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3844..3861
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3899..3954
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3953..3972)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6190..6208)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6276..6380
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6381..6449,6450..7241)
                     /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      7412..8000
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7901..7920
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8154..8171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8288..8309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8324..8329,8330..8347,8348..8354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8362..8378
                     /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     8367..8389
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8386..8402
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8386..8402
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8673..9052
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9053..9256
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9207..9227
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9253..9277
                     /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 ggccgccctg
      421 agcctgaagg gcatcaaaga gggcgaccct ccccggctga tgtacgacaa caacaaagtg
      481 aaccacggca acaccctgaa ggtgtccaag gacgagctga tgaagcggat cgtgcaggac
      541 gtgtcctacg atatcaacaa cttcatcaac gaccagtatg agaacaacaa taacaacaac
      601 aataataaca ataacaacaa caacaacatg cggcgggaca agaccagcga gaacgtgaaa
      661 gagctgcaga acgacgacac cgtgtacatc aagtccctgc agaacaagcc cctgctgaag
      721 atgaacaaca aggaaaaaaa caacaagatc gagatgaagg acagcaagaa gggcaactac
      781 aacaagtaca agatcaagca caacaacaag gcctacgacg gcggcatcag caacagcaga
      841 gagcagaaca acgacgagaa ctacatgaga gaggaacagc ggggcaacat ggccttcaag
      901 ctgaacaacc tgaacaagaa gaagtcccgg atcaagaaca ataagaacgg caagaaagag
      961 aacaatatcg acattaacaa caatatgatc aagatcaaca acatcaagaa ggacatcccc
     1021 aagatcgaca acgacaacaa caacaacaat aacaatgata acaacaataa taacaacaac
     1081 gataacaaca acaacaacaa taacgacaac aataacaata acaacaatgt ggacggcgac
     1141 gacaacaccg ccatccagaa caagggcccc accaatatcg ccagcaccaa cgagcacaag
     1201 aacatcaaca caaacaatca gaacaaacag aagctgcact ccgagaacca ccacaaaaac
     1261 aagctgaccg agacaagcca caacggcgac gtgatcgaga tcagaatcat cagcgacccc
     1321 ggcttcgtga agtccaacca caagaaaacc gagaagaaca agaaatacga catgaaggaa
     1381 aaggacctgc tgcgcatcaa gaacgggaag cggaacaaag aactgatggg cgtgcccgac
     1441 aagatgagca agcgccgcga ggtggaacgc gagaaggacc acatcaaggg caattctaaa
     1501 aacaagaata agaagaagaa taaaaacaaa aagaaaaaca agaacaagaa aaaaaacaag
     1561 aacaaaaact tcgccatcaa gaccaacgat gagtctaaga aaaataagat cagctacatc
     1621 cagatgtaca ccgacaagaa cggcgccgac gagtacagcc ccaagctgaa tgacgacgag
     1681 acattcgacg aggatatgta cgagggcaat gacgagatcg acagcaatat ggccattaag
     1741 aagatcaacg aggataagaa cctgctcgag ttcggcgacg agggcgtgaa ggatatcggc
     1801 tccaccgaga acgagatgat caaaaacaag aagaacatca tcaagaacag ccggaaggtg
     1861 tacaacctca acctgtacaa ctgcgatctg atcgacgact gggacctgaa ccacgagtac
     1921 atcgacgaag agggcaagct gatcaagctg agcggctacg tgttccagaa catcgtgtcc
     1981 accgacagca tgcccaccgt gaatatcgcc gactggaagc tgcggggcag ctgctcctac
     2041 gacaactaca tctgtggcgc cctgaattac atgaataaca tctacagcaa gggcgaccgc
     2101 gtgatcttcg acggcaagat ctacgaggcc accgccgagg tgtacgagac acccaaagtg
     2161 atggaaaacc tgtggatcga aaagaccaag gactgctacg acttcggcgc gccgtcgacc
     2221 tccatcacgg cctataagag tgagggggag tcagcggagt tctccttccc actcaacctt
     2281 ggagaggaaa gcctgcaggg agagttgaga tggaaggcag agaaggctcc ttcttcccag
     2341 tcctggatca ccttctccct aaagaaccaa aaggtgtctg tgcagaagtc tactagcaac
     2401 cccaagttcc agctgtccga aacgctccca ctcacccttc agatacccca ggtctccctt
     2461 cagtttgctg gttctggcaa cctgaccctg actctggaca gagggatact gtatcaggaa
     2521 gtgaacctgg tggtgatgaa agtgactcag cccgacagca acactttgac ctgtgaggtg
     2581 atgggaccca cctcacccaa gatgagactg atcttgaagc aggagaatca ggaggccagg
     2641 gtctccaggc aggagaaagt gattcaagtg caggcccctg aagcaggggt gtggcaatgt
     2701 ctactgagtg aaggtgaaga ggtcaagatg gactccaaga tccaggtttt atccaaaggg
     2761 ttgaattccg gcggcggctc cggaggcggt accggtggcg gcgacctagc cccacagatg
     2821 cttcgagaac tccaggagac taatgcggcg ctgcaagacg tgagagagct cttgcgacag
     2881 caggtcaagg agatcacctt cctgaagaat acggtgatgg aatgtgacgc ctgcggtcta
     2941 gacagaaacc tgcctccact agcacctctg ggcccacacc cagaaacgct ggtgaaagta
     3001 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3061 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3121 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3181 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3241 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3301 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3361 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3421 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3481 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3541 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3601 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3661 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3721 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggggccc tgactacaaa
     3781 gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga tgacaagggc
     3841 cctcaccacc accatcacca ctaattaatt aaggatcccc cgacctcgac ctctggctaa
     3901 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     3961 aggacatatg ggagggcaaa tcatttggtc gagatccctc ggagatctct agctagagga
     4021 tcgatccccg ccccggacga actaaacctg actacgacat ctctgcccct tcttcgcggg
     4081 gcagtgcatg taatcccttc agttggttgg tacaacttgc caactgggcc ctgttccaca
     4141 tgtgacacgg ggggggacca aacacaaagg ggttctctga ctgtagttga catccttata
     4201 aatggatgtg cacatttgcc aacactgagt ggctttcatc ctggagcaga ctttgcagtc
     4261 tgtggactgc aacacaacat tgcctttatg tgtaactctt ggctgaagct cttacaccaa
     4321 tgctggggga catgtacctc ccaggggccc aggaagacta cgggaggcta caccaacgtc
     4381 aatcagaggg gcctgtgtag ctaccgataa gcggaccctc aagagggcat tagcaatagt
     4441 gtttataagg cccccttgtt aaccctaaac gggtagcata tgcttcccgg gtagtagtat
     4501 atactatcca gactaaccct aattcaatag catatgttac ccaacgggaa gcatatgcta
     4561 tcgaattagg gttagtaaaa gggtcctaag gaacagcgat atctcccacc ccatgagctg
     4621 tcacggtttt atttacatgg ggtcaggatt ccacgagggt agtgaaccat tttagtcaca
     4681 agggcagtgg ctgaagatca aggagcgggc agtgaactct cctgaatctt cgcctgcttc
     4741 ttcattctcc ttcgtttagc taatagaata actgctgagt tgtgaacagt aaggtgtatg
     4801 tgaggtgctc gaaaacaagg tttcaggtga cgcccccaga ataaaatttg gacggggggt
     4861 tcagtggtgg cattgtgcta tgacaccaat ataaccctca caaacccctt gggcaataaa
     4921 tactagtgta ggaatgaaac attctgaata tctttaacaa tagaaatcca tggggtgggg
     4981 acaagccgta aagactggat gtccatctca cacgaattta tggctatggg caacacataa
     5041 tcctagtgca atatgatact ggggttatta agatgtgtcc caggcaggga ccaagacagg
     5101 tgaaccatgt tgttacactc tatttgtaac aaggggaaag agagtggacg ccgacagcag
     5161 cggactccac tggttgtctc taacaccccc gaaaattaaa cggggctcca cgccaatggg
     5221 gcccataaac aaagacaagt ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg
     5281 tcagccccca cacgccgccc tgcggttttg gactgtaaaa taagggtgta ataacttggc
     5341 tgattgtaac cccgctaacc actgcggtca aaccacttgc ccacaaaacc actaatggca
     5401 ccccggggaa tacctgcata agtaggtggg cgggccaaga taggggcgcg attgctgcga
     5461 tctggaggac aaattacaca cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc
     5521 atattcacga ggtcgctgag agcacggtgg gctaatgttg ccatgggtag catatactac
     5581 ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata ggctatccta
     5641 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     5701 atttatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     5761 atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata tgctatccta
     5821 atagagatta gggtagtata tgctatccta atttatatct gggtagcata tactacccaa
     5881 atatctggat agcatatgct atcctaatct atatctgggt agcatatgct atcctaatct
     5941 atatctgggt agcataggct atcctaatct atatctgggt agcatatgct atcctaatct
     6001 atatctgggt agtatatgct atcctaattt atatctgggt agcataggct atcctaatct
     6061 atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct
     6121 gtatccgggt agcatatgct atcctcatga taagctgtca aacatgagaa ttttcttgaa
     6181 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     6241 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6301 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6361 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6421 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6481 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6541 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6601 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     6661 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6721 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6781 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6841 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6901 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     6961 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     7021 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     7081 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     7141 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7201 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     7261 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7321 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7381 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7441 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7501 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     7561 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7621 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7681 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7741 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7801 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7861 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7921 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7981 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     8041 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     8101 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     8161 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     8221 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     8281 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     8341 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     8401 accatgatta cgccaagctc tagctagagg tcgaccaatt ctcatgtttg acagcttatc
     8461 atcgcagatc cgggcaacgt tgttgccatt gctgcaggcg cagaactggt aggtatggaa
     8521 gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata
     8581 aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta
     8641 tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta gttattaata
     8701 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     8761 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     8821 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     8881 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc
     8941 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg
     9001 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     9061 gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     9121 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     9181 atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt
     9241 ctatataagc agagctcgtt tagtgaaccg tcagatcctc actctcttcc gcatcgctgt
     9301 ctgcgagggc cagctgttgg gctcgcggtt gaggacaaac tcttcgcggt ctttccagta
     9361 ctcttggatc ggaaacccgt cggcctccga acggtactcc gccaccgagg gacctgagcg
     9421 agtccgcatc gaccggatcg gaaaacct
//
