LOCUS       sequence-366312-        6706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NSP16 fused to BioID2 and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6706)
  AUTHORS   May DG, Martin-Sancho L, Anschau V, Liu S, Chrisopulos RJ, Scott KL,
            Halfmann CT, Diaz Pena R, Pratt D, Campos AR, Roux KJ
  TITLE     A BioID-Derived Proximity Interactome for SARS-CoV-2 Proteins.
  JOURNAL   Viruses. 2022 Mar 15;14(3). pii: v14030611. doi: 10.3390/v14030611.
  PUBMED    35337019
REFERENCE   2  (bases 1 to 6706)
  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: "Viruses."; 
            date: "2022-03-15"; volume: "14(3). pii: v14030611. doi"; pages: " 
            10.3390/v14030611"
COMMENT     Sequence Label: NSP16-1x-BioID2 pBabe-puro (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..6706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     196..216
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(231..257)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(258..953)
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
     CDS             complement(1894..2310)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(1910..1926)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(1942..1964)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    2511..2710
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             2775..3244
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        3275..3379
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3380..3448,3449..4240)
                     /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     complement(3598..3617)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4400..4988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4889..4908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5142..5159
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             5289..5758
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(5979..6578)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6062..6082)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6559..6578
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(6588..6706,1..211))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6602..6706,1..31)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
       61 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
      121 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
      181 ctggggactt tccacaccct aactgacaca cattccacag ggtcgactca tgcgtaatcc
      241 ggtacatcgt aagggtagct tcttctcagg ctgaactcgc cgctcaggat ctccttgatg
      301 ccctcctcgg tcaggatcag ggcgccgccc ttctcgctca ggcccaccag cttgccggtg
      361 atcttgccct cgcccagcag cttcacctcc tcgcccaggt acagcatctt gctctcgatc
      421 ttgcccttga actccttgaa gctcttctcc ttgaacttct tcaggttctc gctgattctc
      481 ttcagcacct tcagcagcac ctcctttctg tcccagtcct tgccggtgat ctcgtacagg
      541 gtggtggctc tgtccttgat ctcctcgggg atctctctct ggttcacgtt gatgccgatg
      601 cccacgatca gcttgtcctt gctcagctcg cacagcacgc cgctcacctt cttctcctgg
      661 aagtacacgt cgttgggcca cttcaggctg aaggggatct cggtgatctc ctccagggcc
      721 tcgctcacgc tcaggcccag caccaggggc agctgcagca ggttctcgaa ctccttgggg
      781 ttcagcagga agctgaagta caggccgccc tcctggctca gccactttct gcccaggccg
      841 cctctgccct tggtctgtct gtcggccacc agggcggtgc cgtagctcac gttccactcc
      901 ttcagtctct cctgggtgct gtccacctcc ttcagccaga tcaggttctt gaagaattca
      961 gacccgcctc caccaccggt gttgttcacc agcacgtcag aggagatgac cactctgttg
     1021 ttttccctga tgatcaggcg tcccttggac agcaggctca ggatcatgtc gttgatctgt
     1081 ccctccttca ggctcatcac agcggtgcct ctcagcttca gtgggaactt agacatgtcg
     1141 aacagggagt agctagacag ctggataggg ttggtgttgc gccagaagat gtagttggcg
     1201 tgcatcacgt aaccgtcgat ctgttcacgg ggctttccca ggtagttgca gccgatcagg
     1261 aaagcctcgg agctagaggc gttcacgttg gtcacgaaag cggtccacca ggcgaagtgt
     1321 cccatcagct tgtacaggtc agcgttccag gagtgttcgg tgatcttgat agccacagat
     1381 ccgcccaggg ccagcttctg ctggatgaag ccgcagatgt aggtgaagaa accttccttg
     1441 ctgtcgttct ccttggtcac gttcttggtc tttgggtcgt acatgtcaga gatgatcagg
     1501 tcccacttgt tagcggtgtg cacagtagca cagtcgccga tcagggtgct gtcagcatca
     1561 gacacgaagt cgttcaggtc agagtccacc agcagggtac cggtgggcag ccactgtctc
     1621 agcacagcgg tacctggagc cactcccttg tcagaaccag caccgaagtg gatcacgcgc
     1681 atgttgtaag gcacggccag ggtcagggtg ttcaggtact ggcacagctg ggtgtactta
     1741 gccacgttca tcatgatacc cttgggcagg gtggcggagt ctccgtagtt ctgcaggtcg
     1801 cacttctcca gcagcatacg ctgcatcttg tacaggttag gcatagccac tccaggctgc
     1861 caagcctggg aagacatggt ggcggatccg gccggcgcct agagaaggag tgagggctgg
     1921 ataaagggag gatcgaggcg gggtcgaacg aggaggttca agggggagag acggggcgga
     1981 tggaggaaga ggaggcggag gcttagggtg tacaaagggc ttgacccagg gaggggggtc
     2041 aaaagccaag gcttcccagg tcacgatgta ggggacctgg tctgggtgtc catgcgggcc
     2101 aggtgaaaag accttgatct taacctgggt gatgaggtct cggttaaagg tgccgtctcg
     2161 cggccatccg acgttaaagg ttggccattc tgcagagcag aaggtaaccc aacgtctctt
     2221 cttgacatct accgactggt tgtgagcgat ccgctcgaca tctttccagt gacctaaggt
     2281 caaacttaag ggagtggtaa cagtctggcc ctaattttca gacaaataca gaaacacagt
     2341 cagacagaga caacacagaa cgatgctgca gcgctgcagc agacaagacg cgcggcttcg
     2401 gttccaaacc gaaagcaaaa attcagacgg aggcgggaac tgttttaggt tctcgtctcc
     2461 taccagaacc acatatcctg acggggtcgg attccacatc gactcccttc ctcaggtcgg
     2521 gccacaaaaa cggcccccaa agtccctggg acgtctccca gggttgcggc cgggtgttca
     2581 gaactcgtca gttccaccac gggtccgcca gatacagagc tagttagcta actagtaccg
     2641 acgcaggcgc ataaaatcag tcatagacac tagacaatcg gacagacaca gataagttgc
     2701 tggccagctt gcctcccggt ggtgggtcgg tggtccctgg gcaggggtct cccgatcccg
     2761 gacgagcccc caaatgaaag acccccgctg acgggtagtc aatcactcag aggagaccct
     2821 cccaaggaac agcgagacca caagtcggat gcaactgcaa gagggtttat tggatacacg
     2881 ggtacccggg cgactcagtc aatcggagga ctggcgcccc gagtgagggg ttgtgggctc
     2941 ttttattgag ctcggggagc agaagcgcgc gaacagaagc gagaagcgaa ctgattggtt
     3001 agttcaaata aggcacaggg tcatttcagg tccttggggc accctggaaa catctgatgg
     3061 ttctctagaa actgctgagg gctggaccgc atctggggac catctgttct tggccctgag
     3121 ccggggcagg aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct
     3181 gttcttggcc ctgagccggg gcaggaactg cttaccacag atatcctgtt tggcccatat
     3241 tcaggctgca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3301 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3361 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3421 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3481 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3541 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3601 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3661 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3721 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3781 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3841 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3901 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3961 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4021 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4081 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4141 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4201 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4261 atatatactt tagattgatt tgcggccgca aaaggatcta ggtgaagatc ctttttgata
     4321 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4381 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4441 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4501 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4561 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4621 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4681 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4741 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4801 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4861 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4921 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4981 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5041 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     5101 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5161 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5221 gcaactatgg ccatttaatg taaatactta agaaaaaaaa ccaaattaat tttgatacat
     5281 gctgcatgtg aaagaccccc gctgacgggt agtcaatcac tcagaggaga ccctcccaag
     5341 gaacagcgag accacaagtc ggatgcaact gcaagagggt ttattggata cacgggtacc
     5401 cgggcgactc agtcaatcgg aggactggcg ccccgagtga ggggttgtgg gctcttttat
     5461 tgagctcggg gagcagaagc gcgcgaacag aagcgagaag cgaactgatt ggttagttca
     5521 aataaggcac agggtcattt caggtccttg gggcaccctg gaaacatctg atggttctct
     5581 agaaactgct gagggctgga ccgcatctgg ggaccatctg ttcttggccc tgagccgggg
     5641 caggaactgc ttaccacaga tatcctgttt ggcccatatt cagctgttcc atctgttctt
     5701 ggccctgagc cggggcagga actgcttacc acagatatcc tgtttggccc atattcagct
     5761 gttccatctg ttcctgacct tgatctgaac ttctctattc tcagttatgt atttttccat
     5821 gccttgcaaa atggcgttac ttaagctagc ttgccaaacc tacaggtggg gtctttcatt
     5881 cccccctttt tctggagact aaataaaatc ttttatttta tcgatgcatg gggtcgtgcg
     5941 ctcctttcgg tcgggcgctg cgggtcgtgg ggcgggcgtc aggcaccggg cttgcgggtc
     6001 atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc
     6061 cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg caccccggcg
     6121 cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt gccctggtgg
     6181 tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc
     6241 aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc
     6301 gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc
     6361 accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg
     6421 aagagttctt gcagctcggt gacccgctcg atgtggcggt ccggatcgac ggtgtggcgc
     6481 gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg gacgtcgtcg
     6541 cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg taagcttttt gcaaaagcct
     6601 aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg
     6661 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaat
//
