LOCUS       sequence-403123-        5958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a peptide from the LDL receptor binding site of PCSK9 
            genetically fused to SpyCatcher001 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5958)
  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 5958)
  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-PEED
FEATURES             Location/Qualifiers
     source          1..5958
                     /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(582..602)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(624..641)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             652..674
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    689..713
                     /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(713..732)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(714..732)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(816..835)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1041..1118
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1119..2201
                     /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(1138..1157)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3513..3704
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3698..3720)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3806..3945
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3861..3880
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3960..3977)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4131..4719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4211..4230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4890..5681,5682..5750))
                     /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     5513..5532
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5751..5855)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5923..5941
                     /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 tgctcgcttg acggtggaaa
      301 cgggtgccgt cctcctccgg ctcgaggccg ttaacggtca cttgaccctg ctcgttaacg
      361 gtgaaggtga tcgcggtcgc cacttcataa ccatccggcg ccgcggtttc aacaaaggtg
      421 tatttacccg gatacaggta gaaatccttc acttggccgt cgctaatcca ggtgctgatg
      481 gttttaccgc tgctatcacg cagttccatg gtcgcgcccg ccagttcctt gccatcctcg
      541 tcacgtttgc taaacttaat gtgggtcgcg ctgtcgccgc taccctggaa atacaggttc
      601 tcggtggtcg ggatatcgta gtcgtggtgg tggtggtgat gcatatgtat atctccttct
      661 taaagttaaa caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag
      721 tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc
      781 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
      841 ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc
      901 aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc
      961 ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa
     1021 gggagagcgt cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat
     1081 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg
     1141 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
     1201 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
     1261 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
     1321 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
     1381 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
     1441 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
     1501 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
     1561 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc
     1621 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
     1681 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
     1741 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
     1801 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
     1861 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
     1921 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca
     1981 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
     2041 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa
     2101 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2161 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta
     2221 agttagctca ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca
     2281 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     2341 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
     2401 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
     2461 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
     2521 attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga
     2581 ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg
     2641 ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg
     2701 ctcgcggctc ttaccagcct aacttcgatc attggaccgc tgatcgtcac ggcgatttat
     2761 gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc cctatacctt
     2821 gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac ctgaatggaa
     2881 gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat caattcttgc
     2941 ggagaactgt gaatgcgcaa accaaccctt ggcagaacat atccatcgcg tccgccatct
     3001 ccagcagccg cacgcggcgc atctcgggca gcgttgggtc ctggccacgg gtgcgcatga
     3061 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga
     3121 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct
     3181 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt
     3241 cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg
     3301 gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt
     3361 cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt
     3421 catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca
     3481 tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta
     3541 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg
     3601 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc
     3661 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     3721 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     3781 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     3841 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accattgcgg
     3901 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
     3961 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4021 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4081 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4141 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4201 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4261 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4321 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4381 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4441 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4501 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4561 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4621 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4681 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4741 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4801 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4861 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4921 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4981 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5041 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5101 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5161 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg
     5221 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5281 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5341 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5401 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5461 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
     5521 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5581 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5641 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5701 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5761 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5821 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5881 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5941 ccctttcgtc ttcaagaa
//
