LOCUS       sequence-328137-        6432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SpyCatcher-fused influenza HA ministem with removable HisTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6432)
  AUTHORS   Lampinen V, Heinimaki S, Laitinen OH, Pesu M, Hankaniemi MM, 
            Blazevic V, Hytonen VP
  TITLE     Modular vaccine platform based on the norovirus-like particle.
  JOURNAL   J Nanobiotechnology. 2021 Jan 19;19(1):25. doi: 
            10.1186/s12951-021-00772-0.
  PUBMED    33468139
REFERENCE   2  (bases 1 to 6432)
  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: "J 
            Nanobiotechnology."; date: "2021-01-19"; volume: "19(1):25. doi"; 
            pages: " 10.1186/s12951-021-00772-0"
COMMENT     Sequence Label: pET11b-SpyCatcher-HA2
FEATURES             Location/Qualifiers
     source          1..6432
                     /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(1056..1076)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(1098..1115)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1126..1148
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1163..1187
                     /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(1187..1206)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1188..1206)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1290..1309)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1515..1592
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1593..2675
                     /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(1612..1631)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3987..4178
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4172..4194)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4280..4419
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4335..4354
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4434..4451)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4605..5193)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4685..4704)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5364..6155,6156..6224))
                     /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     5987..6006
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6225..6329)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6397..6415
                     /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 gcttattaca gaaaggtgga cagcagaacc
      301 cattcaccat ctttacgcac atacgcttga ccatcacgcg gcgcttccgg gatgtaaccg
      361 ccaccggtac cctggctatc atgcgcgtcc agggtacgtt cgttctccag cagaaccagc
      421 agttccgcgt tgtaggtcca gatgtccaga aaaccatcgt ccactttctt gttcaggttt
      481 tccatacgtt tctcatcctt gttgaattcc ttgccggtcg cggtgtcttg aatgttcatt
      541 ttctcgataa cggtgttcac cttgttggta ataccgttga tcgcgttctg ggtcgcgcta
      601 cccgcgctgc cgttacgcag accggtaacc atacgcagtt tcgcgctacg cacatacttc
      661 gggctttcac cgttggtggt cgggtgggtg ttctggtacg gcaggctgct gttcgcgcta
      721 ccgtggctat cctccagcag gttcacgctg tgggtaacgg tcacgttctt ttccagaacg
      781 gtatccacgg tgtcctcgag accgttaacg gtcacttggc cctgctcgtt aacggtgaag
      841 gtgatcgcgg tcgccacttc ataaccatcc ggcgccgcgg tttcaacaaa ggtgtatttg
      901 cccggataca ggtagaaatc cttcacttga ccgtcgctaa tccaggtgct gatggttttg
      961 ccgctgctat cacgcagttc catggtcgca cccgccagtt ccttgccatc ctcgtcacgt
     1021 ttgctaaact taatgtgggt cgcgctgtca ccgctgccct ggaaatacag gttctcggtg
     1081 gtcgggatat cgtagtcgtg gtggtggtgg tgatgcatat gtatatctcc ttcttaaagt
     1141 taaacaaaat tatttctaga ggggaattgt tatccgctca caattcccct atagtgagtc
     1201 gtattaattt cgcgggatcg agatctcgat cctctacgcc ggacgcatcg tggccggcat
     1261 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
     1321 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
     1381 cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt
     1441 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga
     1501 gcgtcgagat cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc
     1561 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
     1621 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
     1681 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
     1741 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
     1801 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
     1861 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
     1921 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
     1981 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
     2041 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
     2101 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
     2161 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
     2221 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
     2281 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
     2341 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg
     2401 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
     2461 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
     2521 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
     2581 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     2641 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag
     2701 ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc
     2761 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
     2821 atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt
     2881 cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc
     2941 gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc
     3001 gccggcatgg cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg
     3061 atggccttcc ccattatgat tcttctcgct tccggcggca tcgggatgcc cgcgttgcag
     3121 gccatgctgt ccaggcaggt agatgacgac catcagggac agcttcaagg atcgctcgcg
     3181 gctcttacca gcctaacttc gatcattgga ccgctgatcg tcacggcgat ttatgccgcc
     3241 tcggcgagca catggaacgg gttggcatgg attgtaggcg ccgccctata ccttgtctgc
     3301 ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct cgacctgaat ggaagccggc
     3361 ggcacctcgc taacggattc accactccaa gaattggagc caatcaattc ttgcggagaa
     3421 ctgtgaatgc gcaaaccaac ccttggcaga acatatccat cgcgtccgcc atctccagca
     3481 gccgcacgcg gcgcatctcg ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc
     3541 tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat
     3601 caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa
     3661 caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc
     3721 cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac
     3781 ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc
     3841 gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat
     3901 cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca
     3961 gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg
     4021 cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg
     4081 atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct
     4141 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     4201 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     4261 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     4321 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccatt gcggtgtgaa
     4381 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
     4441 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4501 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4561 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4621 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4681 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4741 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4801 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4861 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4921 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4981 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5041 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5101 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5161 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5221 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5281 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5341 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5401 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5461 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5521 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5581 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5641 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     5701 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5761 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5821 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5881 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5941 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     6001 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6061 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6121 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6181 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6241 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6301 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6361 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     6421 cgtcttcaag aa
//
