LOCUS       sequence-224486-        8708 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 8708)
  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 8708)
  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..8708
                     /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    3159..3214
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3213..3232)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5450..5468)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5536..5640
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5641..5709,5710..6501)
                     /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      6672..7260
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7161..7180
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7414..7431
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7548..7569
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7584..7589,7590..7607,7608..7614)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7622..7638
                     /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     7627..7649
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7646..7662
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7646..7662
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7933..8312
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8313..8516
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8467..8487
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8513..8537
                     /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 cactaataat aaggatcccc cgacctcgac ctctggctaa taaaggaaat ttattttcat
     3181 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3241 tcatttggtc gagatccctc ggagatctct agctagagga tcgatccccg ccccggacga
     3301 actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg taatcccttc
     3361 agttggttgg tacaacttgc caactgggcc ctgttccaca tgtgacacgg ggggggacca
     3421 aacacaaagg ggttctctga ctgtagttga catccttata aatggatgtg cacatttgcc
     3481 aacactgagt ggctttcatc ctggagcaga ctttgcagtc tgtggactgc aacacaacat
     3541 tgcctttatg tgtaactctt ggctgaagct cttacaccaa tgctggggga catgtacctc
     3601 ccaggggccc aggaagacta cgggaggcta caccaacgtc aatcagaggg gcctgtgtag
     3661 ctaccgataa gcggaccctc aagagggcat tagcaatagt gtttataagg cccccttgtt
     3721 aaccctaaac gggtagcata tgcttcccgg gtagtagtat atactatcca gactaaccct
     3781 aattcaatag catatgttac ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa
     3841 gggtcctaag gaacagcgat atctcccacc ccatgagctg tcacggtttt atttacatgg
     3901 ggtcaggatt ccacgagggt agtgaaccat tttagtcaca agggcagtgg ctgaagatca
     3961 aggagcgggc agtgaactct cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc
     4021 taatagaata actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg
     4081 tttcaggtga cgcccccaga ataaaatttg gacggggggt tcagtggtgg cattgtgcta
     4141 tgacaccaat ataaccctca caaacccctt gggcaataaa tactagtgta ggaatgaaac
     4201 attctgaata tctttaacaa tagaaatcca tggggtgggg acaagccgta aagactggat
     4261 gtccatctca cacgaattta tggctatggg caacacataa tcctagtgca atatgatact
     4321 ggggttatta agatgtgtcc caggcaggga ccaagacagg tgaaccatgt tgttacactc
     4381 tatttgtaac aaggggaaag agagtggacg ccgacagcag cggactccac tggttgtctc
     4441 taacaccccc gaaaattaaa cggggctcca cgccaatggg gcccataaac aaagacaagt
     4501 ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca cacgccgccc
     4561 tgcggttttg gactgtaaaa taagggtgta ataacttggc tgattgtaac cccgctaacc
     4621 actgcggtca aaccacttgc ccacaaaacc actaatggca ccccggggaa tacctgcata
     4681 agtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac aaattacaca
     4741 cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga ggtcgctgag
     4801 agcacggtgg gctaatgttg ccatgggtag catatactac ccaaatatct ggatagcata
     4861 tgctatccta atctatatct gggtagcata ggctatccta atctatatct gggtagcata
     4921 tgctatccta atctatatct gggtagtata tgctatccta atttatatct gggtagcata
     4981 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     5041 tgctatccta atctgtatcc gggtagcata tgctatccta atagagatta gggtagtata
     5101 tgctatccta atttatatct gggtagcata tactacccaa atatctggat agcatatgct
     5161 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agcataggct
     5221 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     5281 atcctaattt atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     5341 atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct
     5401 atcctcatga taagctgtca aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata
     5461 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     5521 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5581 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5641 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5701 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5761 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5821 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5881 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5941 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6001 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6061 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6121 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6181 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6241 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6301 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6361 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6421 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6481 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6541 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6601 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6661 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6721 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6781 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6841 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6901 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6961 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7021 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7081 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7141 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7201 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7261 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7321 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     7381 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     7441 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     7501 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     7561 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     7621 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     7681 tagctagagg tcgaccaatt ctcatgtttg acagcttatc atcgcagatc cgggcaacgt
     7741 tgttgccatt gctgcaggcg cagaactggt aggtatggaa gatctataca ttgaatcaat
     7801 attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt ggctattggc
     7861 cattgcatac gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat
     7921 gaccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     7981 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     8041 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     8101 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     8161 tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta
     8221 aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt
     8281 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg
     8341 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     8401 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc
     8461 cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
     8521 tagtgaaccg tcagatcctc actctcttcc gcatcgctgt ctgcgagggc cagctgttgg
     8581 gctcgcggtt gaggacaaac tcttcgcggt ctttccagta ctcttggatc ggaaacccgt
     8641 cggcctccga acggtactcc gccaccgagg gacctgagcg agtccgcatc gaccggatcg
     8701 gaaaacct
//
