LOCUS       sequence-313213-        6312 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Newcastle disease virus HN protein (47-575) with 
            C-terminal SpyTag fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6312)
  AUTHORS   Rahikainen R, Rijal P, Tan TK, Wu HJ, Andersson AC, Barrett JR, 
            Bowden TA, Draper SJ, Townsend AR, Howarth M
  TITLE     Overcoming Symmetry Mismatch in Vaccine Nanoassembly through 
            Spontaneous Amidation.
  JOURNAL   Angew Chem Int Ed Engl. 2020 Sep 4. doi: 10.1002/anie.202009663.
  PUBMED    32886840
REFERENCE   2  (bases 1 to 6312)
  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: "Angew Chem 
            Int Ed Engl. 2020 Sep 4. doi: 10.1002/anie.202009663."
COMMENT     Sequence Label: NDV-HN-SpyTag
FEATURES             Location/Qualifiers
     source          1..6312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1761..1770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3480..3518
                     /codon_start=1
                     /product="peptide derived from the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyTag
                     /note="forms a covalent isopeptide bond with the SpyCatcher
                     protein (Zakeri et al., 2012)"
                     /translation="AHIVMVDAYKPTK"
     CDS             3537..3554
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3666..3685)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3731..3786
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3785..3804)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4147..4163)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4147..4163)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4160..4182)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4171..4187
                     /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)."
     promoter        complement(join(4195..4201,4202..4219,4220..4225))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4240..4261
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4390..4407)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4561..5149)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4641..4660)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5320..6111,6112..6180))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5943..5962
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6181..6285)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg grgggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgcggccgt
     1741 ctcaggccga attcaagctt gccaccatgg ggatccttcc cagccctggg atgcctgcgc
     1801 tgctctccct cgtgagcctt ctctccgtgc tgctgatggg ttgcgtagct gaaaccggta
     1861 gcatgggcgc cagcacacct tctgatctcg tgggcatccc caccagaatc agcagagccg
     1921 aggaaaagat cacaagcgcc ctgggcagca accaggacgt ggtggacaga atctacaagc
     1981 aggtcgccct cgagagccct ctggcactgc tgaataccga gacaaccatc atgaacgcca
     2041 tcaccagcct gagctaccag atcaacggcg ctgccaacaa ttctggctgg ggcgctccta
     2101 tccacgatcc tgatttcatc ggcggcatcg gcaaagaact gatcgtggac aatgccagcg
     2161 acgtgaccag cttctacccc agcgcctttc aagagcacct gaacttcatc cccgctccta
     2221 caacaggcag cggctgcaca agaatcccca gcttcgatat gagcgccaca cactactgct
     2281 acacccacaa cgtgatcctg agcggctgta gggatcacag ccacagccat cagtatctgg
     2341 ccctgggcgt gctgagaaca accgccacag gccggatctt tttcagcacc ctgagatcca
     2401 tcagcctgga cgacacccag aaccggaagt cctgtagcgt gtcagccaca cctctgggct
     2461 gcgatatgct gtgcagcaaa gtgaccgaga cagaggaaga ggactacaac agcgccgtgc
     2521 ctacactgat ggcccatggc agactgggct ttgacggcca gtaccacgag aaggacctgg
     2581 acgtgacaac cctgttcgag gactgggtcg ccaactatcc tggtgttggc ggcggaagct
     2641 tcatcgacgg cagagtgtgg tttagcgtgt acggcggcct gaagcctaac agccctagcg
     2701 atacagtcca agagggcaaa tacgtgatct ataagcggta caacgacaca tgccccgacg
     2761 agcaggatta ccagatccgg atggctaaga gcagctacaa gcccggcaga ttcggcggca
     2821 agagaatcca gcaggccatc ctgagcatca aggtgtccac aagcctgggc gaagatcccg
     2881 tgctgacagt gcctcctaac accgtgactc tgatgggcgc cgagggcaga atcctgacag
     2941 tgggcacaag ccactttctg taccagcggg gcagcagcta cttcagccct gctctgctgt
     3001 accccatgac cgtgtccaac aagaccgcca cactgcacag cccctacacc ttcaacgcct
     3061 ttaccagacc tggcagcatc ccctgtcagg cctctgccag atgtcccaat agctgtgtga
     3121 ccggcgtgta cacagacccc tatcctctga tcttctaccg gaatcacacc ctgaggggcg
     3181 tgttcggcac aatgctggat agcgaacagg ccagactgaa ccctgcctcc gccgtgtttg
     3241 atagcaccag cagatcccgg atcaccagag tgtctagcag cagcacaaag gccgcctata
     3301 ccaccagcac ctgtttcaag gtggtcaaga caaacaagac ctactgcctg tctatcgccg
     3361 agatcagcaa caccctgttt ggcgagttca gaatcgtgcc cctgctggtg gaaatcctga
     3421 agaacgatgg cgtgcgcgag gccagaagcg gaggctccgg agggtcaggc ggatctgggg
     3481 ctcatattgt tatggttgat gcttacaagc caactaaggg aagcgggggt accaagcacc
     3541 accatcacca tcactaatga tcactcgaga ctagtatcgc gataattcac tcctcaggtg
     3601 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     3661 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     3721 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     3781 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     3841 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     3901 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     3961 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     4021 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     4081 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta
     4141 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4201 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4261 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatcga
     4321 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4381 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4441 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4501 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4561 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4621 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4681 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4741 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4801 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4861 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4921 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4981 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5041 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5101 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5161 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5221 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5281 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5341 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5401 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5461 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5521 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5581 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5641 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5701 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5761 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5821 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5881 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5941 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6001 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6061 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6121 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6181 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6241 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6301 agtgccacct gg
//
