LOCUS       sequence-364477-        7470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NSP14 fused to BioID2 and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7470)
  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 7470)
  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: NSP14-1x-BioID2 pBabe-puro (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..7470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1369..1378
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2986..3681
                     /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             3682..3708
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3723..3743)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3728..4057
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3908..4043
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4067..4666
                     /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(4067..4086)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4563..4583
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5563..5580)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5734..6322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5814..5833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6482..7273,7274..7342))
                     /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     7105..7124
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7343..7447)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccgc caccatggct
     1381 gagaacgtga ccggtctgtt caaggactgc tctaaggtca tcactggtct gcacccaact
     1441 caggctccaa cccacctgtc tgtggacacc aagttcaaga ccgaaggact gtgcgtggac
     1501 atccccggta tccctaagga catgacctac cgcaggctga tctccatgat gggcttcaag
     1561 atgaactacc aggtgaacgg ataccctaac atgttcatca ccagagagga agctatccgc
     1621 cacgtgaggg cctggatcgg tttcgacgtg gaaggttgcc acgctaccag agaagctgtg
     1681 ggtaccaacc tgccactgca gctgggcttc tctaccggag tgaacctggt ggctgtgcct
     1741 accggctacg tggacacccc aaacaacacc gacttctcta gagtgagcgc caagccccct
     1801 ccaggagacc agttcaagca cctgatcccc ctgatgtaca agggtctgcc ttggaacgtg
     1861 gtgcgcatca agatcgtgca gatgctgtcc gacaccctga agaacctgtc tgacagggtg
     1921 gtgttcgtgc tgtgggctca cggcttcgag ctgaccagca tgaagtactt cgtgaagatc
     1981 ggacccgaac gtacctgctg cctgtgcgac agacgtgcca cctgcttctc caccgcctct
     2041 gacacctacg cttgctggca ccactccatc ggattcgact acgtgtacaa ccctttcatg
     2101 atcgacgtgc agcagtgggg attcaccggt aacctgcagt ccaaccacga cctgtactgc
     2161 caggtgcacg gtaacgccca cgtggcttct tgcgacgcta tcatgaccag atgcctggcc
     2221 gtgcacgagt gcttcgtgaa gcgtgtggac tggaccatcg agtaccctat catcggcgac
     2281 gaactgaaga tcaacgccgc ttgcagaaag gtgcagcaca tggtggtgaa ggccgctctg
     2341 ctggctgaca agttcccagt gctgcacgac atcggaaacc caaaggctat caagtgcgtg
     2401 ccccaggccg acgtggagtg gaagttctac gacgctcagc cctgctccga caaggcctac
     2461 aagatcgagg aactgttcta ctcttacgct acccacagcg acaagttcac cgacggcgtg
     2521 tgcctgttct ggaactgcaa cgtggacagg tacccagcca actctatcgt gtgcagattc
     2581 gacacccgtg tgctgagcaa cctgaacctg cccggatgcg acggcggatc cctgtacgtg
     2641 aacaagcacg ccttccacac ccctgctttc gacaagtctg ccttcgtgaa cctgaagcag
     2701 ctgcctttct tctactacag cgactcccca tgcgagagcc acggcaagca ggtggtgtcc
     2761 gacatcgact acgtgcccct gaagagcgcc acctgcatca cccgctgcaa cctgggcggc
     2821 gccgtgtgca gacaccacgc caacgaatac aggctgtacc tggacgctta caacatgatg
     2881 atcagcgccg gcttctccct gtgggtgtac aagcagttcg acacctacaa cctgtggaac
     2941 accttcacca ggctgcagac cggtggtgga ggcgggtctg aattcttcaa gaacctgatc
     3001 tggctgaagg aggtggacag cacccaggag agactgaagg agtggaacgt gagctacggc
     3061 accgccctgg tggccgacag acagaccaag ggcagaggcg gcctgggcag aaagtggctg
     3121 agccaggagg gcggcctgta cttcagcttc ctgctgaacc ccaaggagtt cgagaacctg
     3181 ctgcagctgc ccctggtgct gggcctgagc gtgagcgagg ccctggagga gatcaccgag
     3241 atccccttca gcctgaagtg gcccaacgac gtgtacttcc aggagaagaa ggtgagcggc
     3301 gtgctgtgcg agctgagcaa ggacaagctg atcgtgggca tcggcatcaa cgtgaaccag
     3361 agagagatcc ccgaggagat caaggacaga gccaccaccc tgtacgagat caccggcaag
     3421 gactgggaca gaaaggaggt gctgctgaag gtgctgaaga gaatcagcga gaacctgaag
     3481 aagttcaagg agaagagctt caaggagttc aagggcaaga tcgagagcaa gatgctgtac
     3541 ctgggcgagg aggtgaagct gctgggcgag ggcaagatca ccggcaagct ggtgggcctg
     3601 agcgagaagg gcggcgccct gatcctgacc gaggagggca tcaaggagat cctgagcggc
     3661 gagttcagcc tgagaagaag ctacccttac gatgtaccgg attacgcatg agtcgaccct
     3721 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3781 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3841 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3901 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3961 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4021 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttaccatga ccgagtacaa
     4081 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     4141 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     4201 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     4261 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     4321 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     4381 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     4441 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     4501 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     4561 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     4621 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga
     4681 cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcgataaaat aaaagatttt
     4741 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     4801 ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag
     4861 atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
     4921 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag
     4981 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     5041 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     5101 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     5161 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag
     5221 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     5281 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgakt gwmtmcmmkw
     5341 mascskskkk cwktyrcats crrcwtgtgg tctcgctgtt ccttgggagg gtctcctctg
     5401 agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa aattaatttg
     5461 gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa tcggccaacg
     5521 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5581 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5641 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5701 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5761 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5821 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5881 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5941 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6001 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6061 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6121 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6181 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6241 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6301 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6361 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6421 ctagatcctt ttgcggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6481 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6541 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6601 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6661 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6721 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6781 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6841 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6901 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6961 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7021 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7081 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7141 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7201 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7261 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7321 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7381 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7441 ttccgcgcac atttccccga aaagtgccac
//
