LOCUS       sequence-403129-        6315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses furin P-domain genetically fused to SpyCatcher001 in 
            bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6315)
  AUTHORS   Lampinen V, Ojanen MJT, Caro FM, Grohn S, Hankaniemi MM, Pesu M, 
            Hytonen VP
  TITLE     Experimental VLP vaccine displaying a furin antigen elicits 
            production of autoantibodies and is well tolerated in mice.
  JOURNAL   Nanoscale Adv. 2024 Oct 9;6(24):6239-52. doi: 10.1039/d4na00483c.
  PUBMED    39430302
REFERENCE   2  (bases 1 to 6315)
  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"; doi: "10"; journalName: 
            "Nanoscale Adv"; date: "2024-10-9- 9"; volume: "6"; issue: "24"; 
            pages: "6239-52"
COMMENT     Sequence Label: SpyCatcher-P-Domain
FEATURES             Location/Qualifiers
     source          1..6315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(939..959)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(981..998)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1009..1031
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1046..1070
                     /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     complement(1070..1089)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1071..1089)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1173..1192)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1398..1475
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1476..2558
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1495..1514)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3870..4061
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4055..4077)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4163..4302
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4218..4237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4317..4334)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4488..5076)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4568..4587)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5247..6038,6039..6107))
                     /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"
     primer_bind     5870..5889
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6108..6212)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6280..6298
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcttattatt ccggcgcggt gccatacaga
      301 accagggtga atttggtcag ggtgccgtag ttgttcgctt cgctggtgtt ttcaatctcc
      361 agcacccatt cacccgccgg atcctcatcc cagctatggg tggtcataaa cgcccagtcg
      421 ttgaaaccat ccgcgctata gtcatgcgga cgcgccgcca gcagggtgct acgggtaccc
      481 atcgggctaa tcaggtggat cgccaggtca ccacgacggt tgtagctcag ggtcagacgc
      541 gcctgaacgt gttccagacg ggtgatgtgg ttcggttcac ccaggcacgc ggtaaccgct
      601 ttacgcactt ccagacgttt accaatatcc ttcggttcca ccaggatctc aacaatgcac
      661 ttacgctgcg gcgcaacctc gaggccgtta acggtcactt gaccctgctc gttaacggtg
      721 aaggtgatcg cggtcgccac ttcataacca tccggcgccg cggtttcaac aaaggtgtat
      781 ttacccggat acaggtagaa atccttcact tggccgtcgc taatccaggt gctgatggtt
      841 ttaccgctgc tatcacgcag ttccatggtc gcgcccgcca gttccttgcc atcctcgtca
      901 cgtttgctaa acttaatgtg ggtcgcgctg tcgccgctac cctggaaata caggttctcg
      961 gtggtcggga tatcgtagtc gtggtggtgg tggtgatgca tatgtatatc tccttcttaa
     1021 agttaaacaa aattatttct agaggggaat tgttatccgc tcacaattcc cctatagtga
     1081 gtcgtattaa tttcgcggga tcgagatctc gatcctctac gccggacgca tcgtggccgg
     1141 catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga
     1201 agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg
     1261 ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc
     1321 ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg
     1381 agagcgtcga gatcccggac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat
     1441 agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg
     1501 tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc
     1561 acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc
     1621 ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct
     1681 ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc
     1741 aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag
     1801 cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg
     1861 atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg
     1921 atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac
     1981 tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat
     2041 taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc
     2101 aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa
     2161 ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga
     2221 tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct
     2281 cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca
     2341 tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc
     2401 agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca
     2461 ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     2521 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt
     2581 tagctcactc attaggcacc gggatctcga ccgatgccct tgagagcctt caacccagtc
     2641 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt
     2701 atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc
     2761 tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc
     2821 ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt
     2881 atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc
     2941 tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg
     3001 caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca aggatcgctc
     3061 gcggctctta ccagcctaac ttcgatcatt ggaccgctga tcgtcacggc gatttatgcc
     3121 gcctcggcga gcacatggaa cgggttggca tggattgtag gcgccgccct ataccttgtc
     3181 tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg aatggaagcc
     3241 ggcggcacct cgctaacgga ttcaccactc caagaattgg agccaatcaa ttcttgcgga
     3301 gaactgtgaa tgcgcaaacc aacccttggc agaacatatc catcgcgtcc gccatctcca
     3361 gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg
     3421 tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg
     3481 aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag
     3541 caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag
     3601 cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa
     3661 cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc
     3721 gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat
     3781 catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga
     3841 acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca
     3901 tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg
     3961 cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca
     4021 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     4081 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     4141 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
     4201 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc attgcggtgt
     4261 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
     4321 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4381 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4441 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4501 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4561 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4621 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4681 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4741 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4801 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4861 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4921 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4981 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5041 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5101 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5161 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5221 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5281 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5341 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5401 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5461 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5521 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca
     5581 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5641 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5701 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5761 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5821 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     5881 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5941 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6001 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6061 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6121 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6181 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6241 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6301 tttcgtcttc aagaa
//
