LOCUS       sequence-208750-        8710 bp    DNA     circular SYN 15-DEC-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 8710)
  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 8710)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 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: CHT1-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8710
                     /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             2242..3027
                     /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             3034..3099
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3106..3123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3161..3216
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3215..3234)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5452..5470)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5538..5642
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5643..5711,5712..6503)
                     /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      6674..7262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7163..7182
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7416..7433
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7550..7571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7586..7591,7592..7609,7610..7616)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7624..7640
                     /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     7629..7651
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7648..7664
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7648..7664
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7935..8314
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8315..8518
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8469..8489
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8515..8539
                     /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 ggccgcccat
      421 agagccagac ccggcgagag ccggaagaac cccagagaga tcatcaagac cttcaaagag
      481 agcggcaagg gcatcatcca gggctactac ccctcctggg tgtcctacaa ccacaacctg
      541 aaggacctga accccaatct gaacgtggtg cacatgagct tcgccaagat ggacctgagc
      601 tacgacagca tcgagagcat cgtgggcagc cctctgctgt tcaagagcct gatcggcctc
      661 gagtacatcg gcctgaacga gtacttcaac gacgccatga acctgcggaa ggccaggccc
      721 gacatcatca tgctgctgag cctgggcggc gagacatacc accccagcag cttcgacagc
      781 gccctgaacg ccgtggaaaa gatcgccaac ctggtggacg agctgggctt cgacggcatc
      841 gacgtggact acgagcccaa cggcgccttc gacggactga acgacaaaga gaaggccgac
      901 ttcttcgtgc agtacgtgac caagctgcgc gagtacatgt gcgacgacaa gctgatctcc
      961 atcagccaga gcagcaatgg cgccctgagc tgcatcggct tcaacgatcc caagaaaatc
     1021 tgcatggacg acgaggcccc ttacaacagc aagtacttta acaagcccga cgtgaagaaa
     1081 gagctgctga gagccgccca gatggccagt gccggcggag ccatctacct gatgaacaat
     1141 ctgaaggata tgatcgacat ggtgttcgtg cagaccttta actacgccaa cagcgccgac
     1201 agcaccgtga tgaaggaact gtacgacagc tacgcctact acggcaagaa atacgactac
     1261 gtgatcatca tgggcttcac cctgatgttc cccagcaccc ctttcaaccc caacgacaag
     1321 atgctggtca agtccatcgg cgacttcgtg aaaaccgaga acaagctgaa caagcgggcc
     1381 gacggcttcg gcctgtggtc cctgtctagc gacaacgccg cccacaacga gcagctggcc
     1441 atcgagtact ttgtggaaag cctgcacggc gcgccgtcga cctccatcac ggcctataag
     1501 agtgaggggg agtcagcgga gttctccttc ccactcaacc ttggagagga aagcctgcag
     1561 ggagagttga gatggaaggc agagaaggct ccttcttccc agtcctggat caccttctcc
     1621 ctaaagaacc aaaaggtgtc tgtgcagaag tctactagca accccaagtt ccagctgtcc
     1681 gaaacgctcc cactcaccct tcagataccc caggtctccc ttcagtttgc tggttctggc
     1741 aacctgaccc tgactctgga cagagggata ctgtatcagg aagtgaacct ggtggtgatg
     1801 aaagtgactc agcccgacag caacactttg acctgtgagg tgatgggacc cacctcaccc
     1861 aagatgagac tgatcttgaa gcaggagaat caggaggcca gggtctccag gcaggagaaa
     1921 gtgattcaag tgcaggcccc tgaagcaggg gtgtggcaat gtctactgag tgaaggtgaa
     1981 gaggtcaaga tggactccaa gatccaggtt ttatccaaag ggttgaattc cggcggcggc
     2041 tccggaggcg gtaccggtgg cggcgaccta gccccacaga tgcttcgaga actccaggag
     2101 actaatgcgg cgctgcaaga cgtgagagag ctcttgcgac agcaggtcaa ggagatcacc
     2161 ttcctgaaga atacggtgat ggaatgtgac gcctgcggtc tagacagaaa cctgcctcca
     2221 ctagcacctc tgggcccaca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2281 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2341 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2401 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2461 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2521 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2581 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2641 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2701 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2761 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2821 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     2881 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     2941 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3001 ataggtgcct cactgattaa gcattggggc cctgactaca aagaccatga cggtgattat
     3061 aaagatcatg acatcgatta caaggatgac gatgacaagg gccctcacca ccaccatcac
     3121 cactaattaa ttaaggatcc cccgacctcg acctctggct aataaaggaa atttattttc
     3181 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     3241 aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc cgccccggac
     3301 gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca tgtaatccct
     3361 tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac ggggggggac
     3421 caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg tgcacatttg
     3481 ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact gcaacacaac
     3541 attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg gacatgtacc
     3601 tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag gggcctgtgt
     3661 agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa ggcccccttg
     3721 ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc cagactaacc
     3781 ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta gggttagtaa
     3841 aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt ttatttacat
     3901 ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt ggctgaagat
     3961 caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct ccttcgttta
     4021 gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc tcgaaaacaa
     4081 ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt ggcattgtgc
     4141 tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg taggaatgaa
     4201 acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg taaagactgg
     4261 atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg caatatgata
     4321 ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat gttgttacac
     4381 tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc actggttgtc
     4441 tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa acaaagacaa
     4501 gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc cacacgccgc
     4561 cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta accccgctaa
     4621 ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg aatacctgca
     4681 taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg acaaattaca
     4741 cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac gaggtcgctg
     4801 agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat ctggatagca
     4861 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4921 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     4981 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     5041 tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat tagggtagta
     5101 tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg atagcatatg
     5161 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagcatagg
     5221 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     5281 ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     5341 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg
     5401 ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag ggcctcgtga
     5461 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5521 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5581 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5641 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5701 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5761 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5821 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5881 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5941 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6001 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6061 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6121 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6181 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6241 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6301 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6361 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6421 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6481 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6541 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6601 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6661 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6721 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6781 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6841 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6901 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6961 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7021 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7081 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7141 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7201 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7261 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7321 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     7381 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     7441 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     7501 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     7561 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     7621 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     7681 tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga tccgggcaac
     7741 gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata cattgaatca
     7801 atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata ttggctattg
     7861 gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat
     7921 atgaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     7981 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     8041 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     8101 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     8161 cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg
     8221 taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca
     8281 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacaccaa
     8341 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     8401 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta ataaccccgc
     8461 cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     8521 tttagtgaac cgtcagatcc tcactctctt ccgcatcgct gtctgcgagg gccagctgtt
     8581 gggctcgcgg ttgaggacaa actcttcgcg gtctttccag tactcttgga tcggaaaccc
     8641 gtcggcctcc gaacggtact ccgccaccga gggacctgag cgagtccgca tcgaccggat
     8701 cggaaaacct
//
