LOCUS       sequence-224561-        8180 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 8180)
  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 8180)
  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: CYP19B-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8180
                     /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             1714..2499
                     /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             2506..2571
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2578..2595
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2631..2686
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2685..2704)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4922..4940)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5008..5112
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5113..5181,5182..5973)
                     /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      6144..6732
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6633..6652
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6886..6903
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7020..7041
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7056..7061,7062..7079,7080..7086)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7094..7110
                     /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     7099..7121
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7118..7134
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7118..7134
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7405..7784
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7785..7988
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7939..7959
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7985..8009
                     /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 ggccgccgcc
      421 gaggaacacg agatcaccca caagacctac ttcgacatca ccatcgacga caagcccctg
      481 ggccggatcg tgttcggcct gtacggcaag gtggccccta agaccgtgga aaacttcgtg
      541 tccatctgca agggcaccgt ggtggacggc aagatgctgc actacaccaa cagcatcttc
      601 caccggatca tccccaactt catggcccag ggcggcgata tcaccaactt caacggcgct
      661 ggcggcctga gcatctacgg caagaagttc gaggacgaga acttcaaagt gaaccacgcc
      721 aagcggggcc tgctgagcat ggccaacgcc ggcaagaaca ccaacggcgc ccagttcttc
      781 atcctgttca tccccacccc ttggctggac ggccggcacg tggtgttcgg cgaggtggtg
      841 gaaggcctgg acaagctggt gcacatcgag gccgtgggca ccgacagcgg cgagcctctg
      901 aagcgggtgc tggtcaaaga gtccggcgag ctgcctctcg gcgcgccgtc gacctccatc
      961 acggcctata agagtgaggg ggagtcagcg gagttctcct tcccactcaa ccttggagag
     1021 gaaagcctgc agggagagtt gagatggaag gcagagaagg ctccttcttc ccagtcctgg
     1081 atcaccttct ccctaaagaa ccaaaaggtg tctgtgcaga agtctactag caaccccaag
     1141 ttccagctgt ccgaaacgct cccactcacc cttcagatac cccaggtctc ccttcagttt
     1201 gctggttctg gcaacctgac cctgactctg gacagaggga tactgtatca ggaagtgaac
     1261 ctggtggtga tgaaagtgac tcagcccgac agcaacactt tgacctgtga ggtgatggga
     1321 cccacctcac ccaagatgag actgatcttg aagcaggaga atcaggaggc cagggtctcc
     1381 aggcaggaga aagtgattca agtgcaggcc cctgaagcag gggtgtggca atgtctactg
     1441 agtgaaggtg aagaggtcaa gatggactcc aagatccagg ttttatccaa agggttgaat
     1501 tccggcggcg gctccggagg cggtaccggt ggcggcgacc tagccccaca gatgcttcga
     1561 gaactccagg agactaatgc ggcgctgcaa gacgtgagag agctcttgcg acagcaggtc
     1621 aaggagatca ccttcctgaa gaatacggtg atggaatgtg acgcctgcgg tctagacaga
     1681 aacctgcctc cactagcacc tctgggccca cacccagaaa cgctggtgaa agtaaaagat
     1741 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1801 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1861 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1921 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1981 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2041 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2101 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2161 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     2221 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2281 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2341 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2401 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2461 cagatcgctg agataggtgc ctcactgatt aagcattggg gccctgacta caaagaccat
     2521 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gggccctcac
     2581 caccaccatc accactaata ataaggatcc cccgacctcg acctctggct aataaaggaa
     2641 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     2701 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     2761 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     2821 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     2881 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     2941 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     3001 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     3061 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     3121 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     3181 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     3241 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     3301 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     3361 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     3421 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     3481 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     3541 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     3601 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     3661 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     3721 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     3781 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     3841 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     3901 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     3961 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     4021 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     4081 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     4141 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     4201 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     4261 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     4321 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     4381 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     4441 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4501 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     4561 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     4621 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4681 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4741 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     4801 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     4861 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     4921 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4981 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5041 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5101 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5161 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5221 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5281 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5341 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5401 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5461 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5521 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5581 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5641 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5701 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5761 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5821 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5881 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5941 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6001 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6061 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6121 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6181 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6241 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6301 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6361 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6421 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6481 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6541 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6601 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6661 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6721 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6781 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6841 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6901 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6961 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     7021 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     7081 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     7141 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     7201 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata
     7261 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     7321 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     7381 catgtccaat atgaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa
     7441 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     7501 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     7561 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     7621 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg
     7681 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc
     7741 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     7801 agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     7861 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     7921 ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     7981 gcagagctcg tttagtgaac cgtcagatcc tcactctctt ccgcatcgct gtctgcgagg
     8041 gccagctgtt gggctcgcgg ttgaggacaa actcttcgcg gtctttccag tactcttgga
     8101 tcggaaaccc gtcggcctcc gaacggtact ccgccaccga gggacctgag cgagtccgca
     8161 tcgaccggat cggaaaacct
//
