LOCUS       sequence-364400-        7615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NSP13 fused to BioID2 and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7615)
  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 7615)
  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: NSP13-1x-BioID2 pBabe-puro (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..7615
                     /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             3208..3903
                     /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             3904..3930
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3945..3965)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3950..4279
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4130..4265
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4289..4888
                     /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(4289..4308)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4785..4805
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5109..5578
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5708..5725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5879..6467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5959..5978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6627..7418,7419..7487))
                     /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     7250..7269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7488..7592)
                     /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 caccatggcc
     1381 gtgggcgctt gcgtgctgtg caactctcag accagcctga gatgtggagc ttgcatccgc
     1441 aggcctttcc tgtgctgcaa gtgctgctac gaccacgtga tcagcacctc ccacaagctg
     1501 gtgctgtccg tgaacccata cgtgtgcaac gctcctggtt gtgacgtgac tgacgtgacc
     1561 cagctgtacc tgggcggaat gagctactac tgcaagtccc acaagccccc tatcagcttc
     1621 ccactgtgcg ctaacggcca ggtgttcgga ctgtacaaga acacctgcgt gggctccgac
     1681 aacgtgaccg acttcaacgc catcgctacc tgcgactgga ccaacgccgg agactacatc
     1741 ctggctaaca cctgcaccga aaggctgaag ctgttcgccg ctgagaccct gaaggccacc
     1801 gaggaaacct tcaagctgtc ttacggtatc gctaccgtgc gtgaggtgct gtctgaccgc
     1861 gaactgcacc tgtcttggga agtgggcaag ccaagaccac ccctgaaccg taactacgtg
     1921 ttcaccggat acagggtgac caagaacagc aaggtgcaga tcggagagta caccttcgaa
     1981 aagggagact acggtgacgc cgtggtgtac agaggaacca ccacctacaa gctgaacgtg
     2041 ggtgactact tcgtgctgac ctcccacacc gtgatgccac tgtctgctcc caccctggtg
     2101 cctcaggaac actacgtgcg catcaccgga ctgtacccaa ccctgaacat ctccgacgag
     2161 ttcagctcca acgtggccaa ctaccagaaa gtgggcatgc agaagtactc caccctgcag
     2221 ggacctccag gtaccggcaa gtctcacttc gccatcggtc tggctctgta ctacccctct
     2281 gcccgtatcg tgtacactgc ttgttctcac gctgctgtgg atgctctgtg cgagaaggct
     2341 ctgaagtacc tgcctatcga caagtgctcc cgcatcatcc cagccagagc tcgtgtggaa
     2401 tgcttcgaca agttcaaggt gaactctacc ctggagcagt acgtgttctg caccgtgaac
     2461 gccctgcctg aaaccaccgc tgacatcgtg gtgttcgacg agatcagcat ggccaccaac
     2521 tacgacctgt ctgtggtgaa cgctagactg agggctaagc actacgtgta catcggtgac
     2581 ccagctcagc tgcctgctcc tcgtaccctg ctgaccaagg gcaccctgga gccagaatac
     2641 ttcaacagcg tgtgccgcct gatgaagacc atcggtccag acatgttcct gggtacctgc
     2701 cgccgctgcc ctgctgaaat cgtggacacc gtgtctgctc tggtgtacga caacaagctg
     2761 aaggcccaca aggacaagag cgcccagtgc ttcaagatgt tctacaaggg agtgatcacc
     2821 cacgacgtgt ctagcgccat caacaggccc cagatcggtg tggtgcgcga gttcctgacc
     2881 aggaaccccg cctggagaaa ggccgtgttc atctcccctt acaactctca gaacgccgtg
     2941 gcttccaaga tcctgggtct gcctacccag accgtggact cctctcaggg ctctgaatac
     3001 gactacgtga tcttcaccca gaccaccgag accgcccaca gctgcaacgt gaacaggttc
     3061 aacgtggcca tcaccagagc taaagtggga atcctgtgca tcatgagcga cagagacctg
     3121 tacgacaagc tgcagttcac ctccctggag atcccccgca ggaacgtggc taccctgcag
     3181 accggtggtg gaggcgggtc tgaattcttc aagaacctga tctggctgaa ggaggtggac
     3241 agcacccagg agagactgaa ggagtggaac gtgagctacg gcaccgccct ggtggccgac
     3301 agacagacca agggcagagg cggcctgggc agaaagtggc tgagccagga gggcggcctg
     3361 tacttcagct tcctgctgaa ccccaaggag ttcgagaacc tgctgcagct gcccctggtg
     3421 ctgggcctga gcgtgagcga ggccctggag gagatcaccg agatcccctt cagcctgaag
     3481 tggcccaacg acgtgtactt ccaggagaag aaggtgagcg gcgtgctgtg cgagctgagc
     3541 aaggacaagc tgatcgtggg catcggcatc aacgtgaacc agagagagat ccccgaggag
     3601 atcaaggaca gagccaccac cctgtacgag atcaccggca aggactggga cagaaaggag
     3661 gtgctgctga aggtgctgaa gagaatcagc gagaacctga agaagttcaa ggagaagagc
     3721 ttcaaggagt tcaagggcaa gatcgagagc aagatgctgt acctgggcga ggaggtgaag
     3781 ctgctgggcg agggcaagat caccggcaag ctggtgggcc tgagcgagaa gggcggcgcc
     3841 ctgatcctga ccgaggaggg catcaaggag atcctgagcg gcgagttcag cctgagaaga
     3901 agctaccctt acgatgtacc ggattacgca tgagtcgacc ctgtggaatg tgtgtcagtt
     3961 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4021 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4081 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4141 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4201 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4261 aggcctaggc ttttgcaaaa agcttaccat gaccgagtac aagcccacgg tgcgcctcgc
     4321 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4381 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4441 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     4501 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4561 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4621 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     4681 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     4741 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     4801 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     4861 gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga
     4921 aaggagcgca cgaccccatg catcgataaa ataaaagatt ttatttagtc tccagaaaaa
     4981 ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg
     5041 caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga
     5101 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     5161 agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     5221 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca
     5281 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     5341 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     5401 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
     5461 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     5521 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcaca
     5581 tgcagcatgt atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat
     5641 agttgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5701 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5761 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5821 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5881 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5941 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6001 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6061 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6121 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6181 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6241 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6301 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6361 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6421 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6481 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6541 tgagattatc aaaaaggatc ttcacctaga tccttttgcg gccgcaaatc aatctaaagt
     6601 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6661 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6721 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6781 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6841 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6901 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6961 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7021 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7081 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7141 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7201 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7261 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7321 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7381 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7441 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7501 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7561 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
