LOCUS       sequence-208680-        8869 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 8869)
  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 8869)
  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..8869
                     /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    3320..3375
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3374..3393)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5611..5629)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5697..5801
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5802..5870,5871..6662)
                     /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      6833..7421
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7322..7341
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7575..7592
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7709..7730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7745..7750,7751..7768,7769..7775)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7783..7799
                     /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     7788..7810
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7807..7823
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7807..7823
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8094..8473
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8474..8677
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8628..8648
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8674..8698
                     /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 actaattaat taaggatccc
     3301 ccgacctcga cctctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     3361 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttggt cgagatccct
     3421 cggagatctc tagctagagg atcgatcccc gccccggacg aactaaacct gactacgaca
     3481 tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg gtacaacttg
     3541 ccaactgggc cctgttccac atgtgacacg gggggggacc aaacacaaag gggttctctg
     3601 actgtagttg acatccttat aaatggatgt gcacatttgc caacactgag tggctttcat
     3661 cctggagcag actttgcagt ctgtggactg caacacaaca ttgcctttat gtgtaactct
     3721 tggctgaagc tcttacacca atgctggggg acatgtacct cccaggggcc caggaagact
     3781 acgggaggct acaccaacgt caatcagagg ggcctgtgta gctaccgata agcggaccct
     3841 caagagggca ttagcaatag tgtttataag gcccccttgt taaccctaaa cgggtagcat
     3901 atgcttcccg ggtagtagta tatactatcc agactaaccc taattcaata gcatatgtta
     3961 cccaacggga agcatatgct atcgaattag ggttagtaaa agggtcctaa ggaacagcga
     4021 tatctcccac cccatgagct gtcacggttt tatttacatg gggtcaggat tccacgaggg
     4081 tagtgaacca ttttagtcac aagggcagtg gctgaagatc aaggagcggg cagtgaactc
     4141 tcctgaatct tcgcctgctt cttcattctc cttcgtttag ctaatagaat aactgctgag
     4201 ttgtgaacag taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag
     4261 aataaaattt ggacgggggg ttcagtggtg gcattgtgct atgacaccaa tataaccctc
     4321 acaaacccct tgggcaataa atactagtgt aggaatgaaa cattctgaat atctttaaca
     4381 atagaaatcc atggggtggg gacaagccgt aaagactgga tgtccatctc acacgaattt
     4441 atggctatgg gcaacacata atcctagtgc aatatgatac tggggttatt aagatgtgtc
     4501 ccaggcaggg accaagacag gtgaaccatg ttgttacact ctatttgtaa caaggggaaa
     4561 gagagtggac gccgacagca gcggactcca ctggttgtct ctaacacccc cgaaaattaa
     4621 acggggctcc acgccaatgg ggcccataaa caaagacaag tggccactct tttttttgaa
     4681 attgtggagt gggggcacgc gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa
     4741 ataagggtgt aataacttgg ctgattgtaa ccccgctaac cactgcggtc aaaccacttg
     4801 cccacaaaac cactaatggc accccgggga atacctgcat aagtaggtgg gcgggccaag
     4861 ataggggcgc gattgctgcg atctggagga caaattacac acacttgcgc ctgagcgcca
     4921 agcacagggt tgttggtcct catattcacg aggtcgctga gagcacggtg ggctaatgtt
     4981 gccatgggta gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc
     5041 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     5101 tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc
     5161 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc
     5221 cgggtagcat atgctatcct aatagagatt agggtagtat atgctatcct aatttatatc
     5281 tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg
     5341 tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg
     5401 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg
     5461 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     5521 tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctcatg ataagctgtc
     5581 aaacatgaga attttcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5641 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5701 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5761 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5821 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5881 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5941 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6001 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     6061 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6121 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6181 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6241 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6301 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     6361 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6421 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6481 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6541 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6601 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6661 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6721 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6781 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6841 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6901 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6961 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7021 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7081 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7141 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7201 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7261 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7321 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7381 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7441 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7501 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     7561 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     7621 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     7681 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     7741 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     7801 ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag gtcgaccaat
     7861 tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat tgctgcaggc
     7921 gcagaactgg taggtatgga agatctatac attgaatcaa tattggcaat tagccatatt
     7981 agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct
     8041 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gttgacattg
     8101 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     8161 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     8221 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     8281 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     8341 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     8401 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
     8461 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
     8521 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     8581 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
     8641 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcct
     8701 cactctcttc cgcatcgctg tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa
     8761 ctcttcgcgg tctttccagt actcttggat cggaaacccg tcggcctccg aacggtactc
     8821 cgccaccgag ggacctgagc gagtccgcat cgaccggatc ggaaaacct
//
