LOCUS       sequence-221974-        8867 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 8867)
  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 8867)
  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: GPR180-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8867
                     /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             2401..3186
                     /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             3193..3258
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3265..3282
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3318..3373
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3372..3391)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5609..5627)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5695..5799
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5800..5868,5869..6660)
                     /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      6831..7419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7320..7339
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7573..7590
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7707..7728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7743..7748,7749..7766,7767..7773)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7781..7797
                     /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     7786..7808
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7805..7821
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7805..7821
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8092..8471
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8472..8675
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8626..8646
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8672..8696
                     /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 gtgtaccagg gccagttcta cctgaagaag aacgagaaga tccactacct gaccaagttc
      481 agctgcaaca tcggcacctg tcagttcagc atcaagatga agctgcggga caatatcgcc
      541 gccaagctgc tgaacaacta ctacaaactg aacaacaagg ccctgaaaga gatggaaaac
      601 gacaaggacg acggcaacaa ccataacaac aacaaatacg acgacgataa caacaacaac
      661 aacaatcggg ccaacaagct gctgcacatg gactacgaca acgcctacta cgtgctgagc
      721 aaccaggacc tcgagtacat ccctcagcgg atcaccctga tgcaggacat cgagcagaga
      781 tacgtgaaca agaacgacaa gtgctacgcc ttcggcaatc tgaagaacag ccacatgttc
      841 aacatcccat tctacttcaa gtacagcgac ctggtcaaca gcaagaccta caacctcaag
      901 aacttcatca acaacaagta ctccatgagc ttcaacgcca tccacagcga gctgaaaaac
      961 aacaaactgc tccagagcga gatcctgaac tacggcccca tcctgaatga gaagaactcc
     1021 aactacgcct acaaagagat ctacgccaag accacccacc gggtgcacgt gtggtttctg
     1081 atgttcgacg actgctacga caatttcatc cggaacctga agctgaacat caagaccaag
     1141 ctggcctact gggagtacct gctgcacgcc agcaagaagt tcgacatcag cagcctgaac
     1201 gacatggaca tcaagcagga caccagctac aacgacttcc agaacgagga aaccgagaag
     1261 gacatctaca gcaaagagaa catcaacaat aataataaca ataacatgaa taacaacatg
     1321 aacgagaaaa agaacaaggc catcttctac aagtccttca acttcgacaa gatcaccctc
     1381 ctggaaaacc ccaagcggtt cctgttcctg gaagaacagt tcaccaagtc cgagatcaac
     1441 gatatctcat tcttcctgaa gtactaccag ttcaagaaga tgaagaattt caacaatatg
     1501 tacaacaacc tgtacaagaa cggctactac gagcaggtca tcgactacat ctaccagaat
     1561 gagggcttcc acatcgagta cgaagtgaac atcttccaga ccaacaaagc ccacttcagc
     1621 tacgagggcg cgccgtcgac ctccatcacg gcctataaga gtgaggggga gtcagcggag
     1681 ttctccttcc cactcaacct tggagaggaa agcctgcagg gagagttgag atggaaggca
     1741 gagaaggctc cttcttccca gtcctggatc accttctccc taaagaacca aaaggtgtct
     1801 gtgcagaagt ctactagcaa ccccaagttc cagctgtccg aaacgctccc actcaccctt
     1861 cagatacccc aggtctccct tcagtttgct ggttctggca acctgaccct gactctggac
     1921 agagggatac tgtatcagga agtgaacctg gtggtgatga aagtgactca gcccgacagc
     1981 aacactttga cctgtgaggt gatgggaccc acctcaccca agatgagact gatcttgaag
     2041 caggagaatc aggaggccag ggtctccagg caggagaaag tgattcaagt gcaggcccct
     2101 gaagcagggg tgtggcaatg tctactgagt gaaggtgaag aggtcaagat ggactccaag
     2161 atccaggttt tatccaaagg gttgaattcc ggcggcggct ccggaggcgg taccggtggc
     2221 ggcgacctag ccccacagat gcttcgagaa ctccaggaga ctaatgcggc gctgcaagac
     2281 gtgagagagc tcttgcgaca gcaggtcaag gagatcacct tcctgaagaa tacggtgatg
     2341 gaatgtgacg cctgcggtct agacagaaac ctgcctccac tagcacctct gggcccacac
     2401 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2461 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2521 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2581 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2641 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2701 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2761 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2821 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2881 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2941 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3001 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3061 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3121 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3181 cattggggcc ctgactacaa agaccatgac ggtgattata aagatcatga catcgattac
     3241 aaggatgacg atgacaaggg ccctcaccac caccatcacc actaataata aggatccccc
     3301 gacctcgacc tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     3361 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttggtcg agatccctcg
     3421 gagatctcta gctagaggat cgatccccgc cccggacgaa ctaaacctga ctacgacatc
     3481 tctgcccctt cttcgcgggg cagtgcatgt aatcccttca gttggttggt acaacttgcc
     3541 aactgggccc tgttccacat gtgacacggg gggggaccaa acacaaaggg gttctctgac
     3601 tgtagttgac atccttataa atggatgtgc acatttgcca acactgagtg gctttcatcc
     3661 tggagcagac tttgcagtct gtggactgca acacaacatt gcctttatgt gtaactcttg
     3721 gctgaagctc ttacaccaat gctgggggac atgtacctcc caggggccca ggaagactac
     3781 gggaggctac accaacgtca atcagagggg cctgtgtagc taccgataag cggaccctca
     3841 agagggcatt agcaatagtg tttataaggc ccccttgtta accctaaacg ggtagcatat
     3901 gcttcccggg tagtagtata tactatccag actaacccta attcaatagc atatgttacc
     3961 caacgggaag catatgctat cgaattaggg ttagtaaaag ggtcctaagg aacagcgata
     4021 tctcccaccc catgagctgt cacggtttta tttacatggg gtcaggattc cacgagggta
     4081 gtgaaccatt ttagtcacaa gggcagtggc tgaagatcaa ggagcgggca gtgaactctc
     4141 ctgaatcttc gcctgcttct tcattctcct tcgtttagct aatagaataa ctgctgagtt
     4201 gtgaacagta aggtgtatgt gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa
     4261 taaaatttgg acggggggtt cagtggtggc attgtgctat gacaccaata taaccctcac
     4321 aaaccccttg ggcaataaat actagtgtag gaatgaaaca ttctgaatat ctttaacaat
     4381 agaaatccat ggggtgggga caagccgtaa agactggatg tccatctcac acgaatttat
     4441 ggctatgggc aacacataat cctagtgcaa tatgatactg gggttattaa gatgtgtccc
     4501 aggcagggac caagacaggt gaaccatgtt gttacactct atttgtaaca aggggaaaga
     4561 gagtggacgc cgacagcagc ggactccact ggttgtctct aacacccccg aaaattaaac
     4621 ggggctccac gccaatgggg cccataaaca aagacaagtg gccactcttt tttttgaaat
     4681 tgtggagtgg gggcacgcgt cagcccccac acgccgccct gcggttttgg actgtaaaat
     4741 aagggtgtaa taacttggct gattgtaacc ccgctaacca ctgcggtcaa accacttgcc
     4801 cacaaaacca ctaatggcac cccggggaat acctgcataa gtaggtgggc gggccaagat
     4861 aggggcgcga ttgctgcgat ctggaggaca aattacacac acttgcgcct gagcgccaag
     4921 cacagggttg ttggtcctca tattcacgag gtcgctgaga gcacggtggg ctaatgttgc
     4981 catgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
     5041 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     5101 ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg
     5161 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg
     5221 ggtagcatat gctatcctaa tagagattag ggtagtatat gctatcctaa tttatatctg
     5281 ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta
     5341 gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta
     5401 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta
     5461 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     5521 gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctcatgat aagctgtcaa
     5581 acatgagaat tttcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5641 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5701 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5761 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5821 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5881 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5941 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6001 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     6061 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6121 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6181 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6241 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6301 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     6361 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6421 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6481 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6541 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6601 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6661 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6721 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6781 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6841 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6901 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6961 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7021 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7081 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7141 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7201 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7261 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7321 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7381 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7441 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7501 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7561 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     7621 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     7681 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7741 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7801 cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt cgaccaattc
     7861 tcatgtttga cagcttatca tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc
     7921 agaactggta ggtatggaag atctatacat tgaatcaata ttggcaatta gccatattag
     7981 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
     8041 atcataatat gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat
     8101 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     8161 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     8221 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     8281 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     8341 atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     8401 cccagtacat gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg
     8461 ctattaccat ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact
     8521 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     8581 atcaacggga ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta
     8641 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcctca
     8701 ctctcttccg catcgctgtc tgcgagggcc agctgttggg ctcgcggttg aggacaaact
     8761 cttcgcggtc tttccagtac tcttggatcg gaaacccgtc ggcctccgaa cggtactccg
     8821 ccaccgaggg acctgagcga gtccgcatcg accggatcgg aaaacct
//
