LOCUS Exported 6312 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses Newcastle disease virus HN protein (47-575) with C-terminal SpyTag fusion in mammalian cells. ACCESSION . VERSION . KEYWORDS NDV-HN-SpyTag 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 May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6312 /organism="synthetic DNA construct" /mol_type="other DNA" 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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" 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(4195..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(5320..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 //