LOCUS       sequence-208764-        9154 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 9154)
  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 9154)
  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: TRAP-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9154
                     /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             2686..3471
                     /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             3478..3543
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3550..3567
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3605..3660
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3659..3678)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5896..5914)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5982..6086
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6087..6155,6156..6947)
                     /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      7118..7706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7607..7626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7860..7877
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7994..8015
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8030..8035,8036..8053,8054..8060)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8068..8084
                     /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     8073..8095
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8092..8108
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8092..8108
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8379..8758
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8759..8962
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8913..8933
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8959..8983
                     /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 ggccgccaca
      421 atggcccatc tgggcaacgt gaagtacctc gtgatcgtgt tcctgatctt cttcgacctg
      481 ttcctcgtga acggccggga cgtgcagaac aacatcgtgg acgagatcaa gtaccgcgag
      541 gaagtgtgca acgacgaggt ggacctgtac ctgctgatgg actgcagcgg cagcatccgg
      601 cggcacaact gggtcaacca tgccgtgcct ctggccatga agctgatcca gcagctgaac
      661 ctgaacgaca acgccatcca cctgtacgcc agcgtgttca gcaacaacgc cagagagatc
      721 atcagactgc acagcgacgc cagcaagaac aaagagaagg ccctgatcat catcaagagc
      781 ctgctgagca ccaacctgcc ctacggcaag accaacctgg ccgatgctct gctgcaagtg
      841 cggaagcacc tgaatgaccg gatcaacaga gagaacgcca atcagctggt cgtgatcctg
      901 accgacggca tccccgacag catccaggac agcctgaaag agagccggaa gctgagcgac
      961 cggggcgtga agatcgccgt gtttggcatc ggacagggca tcaacgtggc cttcaaccgg
     1021 ttcctcgtgg gctgccaccc tagcgacggc aagtgcaatc tgtacgccga cagcgcctgg
     1081 gagaacgtga agaatgtgat cggccctttc atgaaggccg tgtgcgtgga agtggaaaag
     1141 accgccagct gcggcgtgtg ggatgagtgg tccccttgca gcgtgacctg cggcaagggc
     1201 accagaagcc ggaagagaga gatcctgcac gagggctgca ccagcgagct gcaggaacag
     1261 tgcgaagagg aacggtgcct gcccaagagg gaacccctgg atgtgcccga cgagcccgag
     1321 gacgatcagc ctagacccag aggcgacaac ttcgccgtgg aaaagcccaa cgagaacatc
     1381 atcgacaaca acccccagga acccagcccc aaccccgaag agggaaaggg cgagaacccc
     1441 aacggcttcg atctggacga gaaccctgag aaccccccca accctcccaa tccccctaac
     1501 cccccaaatc caccaaatcc tcccaaccct gacatccccg agcaggaacc taacatccct
     1561 gaggacagcg agaaagaggt gcccagcgac gtgcccaaga accccgagga tgaccgggaa
     1621 gagaacttcg acatccccaa gaagccagag aacaagcacg acaaccagaa caacctgcct
     1681 aacgacaagt ccgaccggta catcccctac agccccctga gccccaaggt gctggacaac
     1741 gagagaaagc agagcgaccc ccagagccag gacaacaacg gcaacagaca cgtgccaaac
     1801 agcgaggaca gagagacacg gccccacggc cggaacaacg agaatcgggc ctacaaccgg
     1861 aagcacaaca atgcccccaa gcaccctgag cgggaagaac acgaaaaacc tggcgcgccg
     1921 tcgacctcca tcacggccta taagagtgag ggggagtcag cggagttctc cttcccactc
     1981 aaccttggag aggaaagcct gcagggagag ttgagatgga aggcagagaa ggctccttct
     2041 tcccagtcct ggatcacctt ctccctaaag aaccaaaagg tgtctgtgca gaagtctact
     2101 agcaacccca agttccagct gtccgaaacg ctcccactca cccttcagat accccaggtc
     2161 tcccttcagt ttgctggttc tggcaacctg accctgactc tggacagagg gatactgtat
     2221 caggaagtga acctggtggt gatgaaagtg actcagcccg acagcaacac tttgacctgt
     2281 gaggtgatgg gacccacctc acccaagatg agactgatct tgaagcagga gaatcaggag
     2341 gccagggtct ccaggcagga gaaagtgatt caagtgcagg cccctgaagc aggggtgtgg
     2401 caatgtctac tgagtgaagg tgaagaggtc aagatggact ccaagatcca ggttttatcc
     2461 aaagggttga attccggcgg cggctccgga ggcggtaccg gtggcggcga cctagcccca
     2521 cagatgcttc gagaactcca ggagactaat gcggcgctgc aagacgtgag agagctcttg
     2581 cgacagcagg tcaaggagat caccttcctg aagaatacgg tgatggaatg tgacgcctgc
     2641 ggtctagaca gaaacctgcc tccactagca cctctgggcc cacacccaga aacgctggtg
     2701 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2761 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2821 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2881 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2941 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3001 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3061 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3121 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3181 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3241 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3301 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3361 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3421 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gggccctgac
     3481 tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac
     3541 aagggccctc accaccacca tcaccactaa ttaattaagg atcccccgac ctcgacctct
     3601 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     3661 ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag atctctagct
     3721 agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct gccccttctt
     3781 cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac tgggccctgt
     3841 tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt agttgacatc
     3901 cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg agcagacttt
     3961 gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct gaagctctta
     4021 caccaatgct gggggacatg tacctcccag gggcccagga agactacggg aggctacacc
     4081 aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga gggcattagc
     4141 aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct tcccgggtag
     4201 tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa cgggaagcat
     4261 atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct cccaccccat
     4321 gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg aaccatttta
     4381 gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg aatcttcgcc
     4441 tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg aacagtaagg
     4501 tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa aatttggacg
     4561 gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa ccccttgggc
     4621 aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga aatccatggg
     4681 gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc tatgggcaac
     4741 acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg cagggaccaa
     4801 gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag tggacgccga
     4861 cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg gctccacgcc
     4921 aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt ggagtggggg
     4981 cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag ggtgtaataa
     5041 cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac aaaaccacta
     5101 atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg ggcgcgattg
     5161 ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac agggttgttg
     5221 gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat gggtagcata
     5281 tactacccaa atatctggat agcatatgct atcctaatct atatctgggt agcataggct
     5341 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     5401 atcctaattt atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     5461 atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct
     5521 atcctaatag agattagggt agtatatgct atcctaattt atatctgggt agcatatact
     5581 acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca tatgctatcc
     5641 taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc
     5701 taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca taggctatcc
     5761 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc
     5821 taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca tgagaatttt
     5881 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5941 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6001 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6061 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     6121 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6181 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6241 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6301 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     6361 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6421 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6481 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6541 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6601 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     6661 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6721 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6781 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6841 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6901 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6961 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     7021 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7081 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     7141 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     7201 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     7261 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7321 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7381 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7441 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7501 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7561 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7621 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7681 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7741 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     7801 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     7861 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     7921 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     7981 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     8041 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     8101 gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca tgtttgacag
     8161 cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga actggtaggt
     8221 atggaagatc tatacattga atcaatattg gcaattagcc atattagtca ttggttatat
     8281 agcataaatc aatattggct attggccatt gcatacgttg tatctatatc ataatatgta
     8341 catttatatt ggctcatgtc caatatgacc gccatgttga cattgattat tgactagtta
     8401 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     8461 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     8521 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     8581 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc
     8641 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     8701 cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     8761 gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc
     8821 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     8881 tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg
     8941 ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcctcactc tcttccgcat
     9001 cgctgtctgc gagggccagc tgttgggctc gcggttgagg acaaactctt cgcggtcttt
     9061 ccagtactct tggatcggaa acccgtcggc ctccgaacgg tactccgcca ccgagggacc
     9121 tgagcgagtc cgcatcgacc ggatcggaaa acct
//
