LOCUS       sequence-364328-        7389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 NSP4 fused to BioID2 and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7389)
  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 7389)
  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: NSP4-1x-BioID2 pBabe-puro (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..7389
                     /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"
     CDS             2905..3600
                     /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             3601..3627
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3642..3662)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3647..3976
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3827..3962
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3986..4585
                     /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(3986..4005)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4482..4502
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5482..5499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5653..6241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5733..5752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6401..7192,7193..7261))
                     /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     7024..7043
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7262..7366)
                     /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 caccatgaag
     1381 atcgtgaaca actggctgaa gcagctgatc aaggtgaccc tggtgttcct gttcgtggcc
     1441 gctatcttct acctgatcac ccctgtgcac gtgatgagca agcacaccga cttcagctcc
     1501 gagatcatcg gctacaaggc catcgacggc ggagtgacca gagacatcgc ttccaccgac
     1561 acctgcttcg ccaacaagca cgctgacttc gacacctggt tctcccagag aggtggctct
     1621 tacaccaacg acaaggcctg cccactgatc gccgctgtga tcacccgtga agtgggattc
     1681 gtggtgcctg gcctgccagg aaccatcctg agaaccacca acggtgactt cctgcacttc
     1741 ctgccacgcg tgttcagcgc cgtgggcaac atctgctaca ccccttctaa gctgatcgag
     1801 tacaccgact tcgccacctc tgcttgcgtg ctggccgctg aatgcaccat cttcaaggac
     1861 gccagcggaa agccagtgcc ctactgctac gacaccaacg tgctggaggg tagcgtggct
     1921 tacgaatccc tgcgcccaga caccagatac gtgctgatgg acggctccat catccagttc
     1981 cccaacacct acctggaggg aagcgtgcgc gtggtgacca ccttcgactc cgaatactgc
     2041 agacacggaa cctgcgagcg ttccgaagcc ggagtgtgcg tgtctaccag cggtcgctgg
     2101 gtgctgaaca acgactacta caggtctctg cctggcgtgt tctgcggagt ggatgctgtg
     2161 aacctgctga ccaacatgtt caccccactg atccagccca tcggtgctct ggacatctcc
     2221 gcctctatcg tggctggagg tatcgtggcc atcgtggtga cctgcctggc ttactacttc
     2281 atgaggttcc gcagggcctt cggtgaatac agccacgtgg tggctttcaa caccctgctg
     2341 ttcctgatgt ccttcaccgt gctgtgcctg acccccgtgt actctttcct gccaggcgtg
     2401 tacagcgtga tctacctgta cctgaccttc tacctgacca acgacgtgtc tttcctggcc
     2461 cacatccagt ggatggtcat gttcacccct ctggtgcctt tctggatcac catcgcttac
     2521 atcatctgca tctctaccaa gcacttctac tggttcttca gcaactacct gaagagacgt
     2581 gtggtgttca acggcgtgag cttctccacc ttcgaggaag ccgctctgtg caccttcctg
     2641 ctgaacaagg agatgtacct gaagctgcgc agcgacgtgc tgctgcctct gacccagtac
     2701 aacaggtacc tggccctgta caacaagtac aagtacttct ccggcgctat ggacaccacc
     2761 tcttaccgtg aagccgcttg ctgccacctg gccaaggctc tgaacgactt ctctaacagc
     2821 ggatccgacg tgctgtacca gccccctcag acctctatca ccagcgccgt gctgcagacc
     2881 ggtggtggag gcgggtctga attcttcaag aacctgatct ggctgaagga ggtggacagc
     2941 acccaggaga gactgaagga gtggaacgtg agctacggca ccgccctggt ggccgacaga
     3001 cagaccaagg gcagaggcgg cctgggcaga aagtggctga gccaggaggg cggcctgtac
     3061 ttcagcttcc tgctgaaccc caaggagttc gagaacctgc tgcagctgcc cctggtgctg
     3121 ggcctgagcg tgagcgaggc cctggaggag atcaccgaga tccccttcag cctgaagtgg
     3181 cccaacgacg tgtacttcca ggagaagaag gtgagcggcg tgctgtgcga gctgagcaag
     3241 gacaagctga tcgtgggcat cggcatcaac gtgaaccaga gagagatccc cgaggagatc
     3301 aaggacagag ccaccaccct gtacgagatc accggcaagg actgggacag aaaggaggtg
     3361 ctgctgaagg tgctgaagag aatcagcgag aacctgaaga agttcaagga gaagagcttc
     3421 aaggagttca agggcaagat cgagagcaag atgctgtacc tgggcgagga ggtgaagctg
     3481 ctgggcgagg gcaagatcac cggcaagctg gtgggcctga gcgagaaggg cggcgccctg
     3541 atcctgaccg aggagggcat caaggagatc ctgagcggcg agttcagcct gagaagaagc
     3601 tacccttacg atgtaccgga ttacgcatga gtcgaccctg tggaatgtgt gtcagttagg
     3661 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3721 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3781 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3841 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3901 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3961 cctaggcttt tgcaaaaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac
     4021 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4081 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4141 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4201 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     4261 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     4321 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     4381 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     4441 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     4501 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     4561 catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag
     4621 gagcgcacga ccccatgcat cgataaaata aaagatttta tttagtctcc agaaaaaggg
     4681 gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
     4741 ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag gaacagatgg
     4801 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     4861 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     4921 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt
     4981 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     5041 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     5101 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc
     5161 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     5221 ttccttggga gggtctcctc tgagysmktg wmtmcmmkwm ascskskkgc wgttgcatcc
     5281 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
     5341 gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta
     5401 cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5461 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5521 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5581 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5641 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5701 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5761 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5821 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5881 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5941 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6001 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6061 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6121 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6181 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6241 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6301 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccgca
     6361 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6421 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6481 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6541 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6601 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6661 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6721 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6781 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6841 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6901 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6961 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7021 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7081 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7141 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7201 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7261 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7321 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7381 aagtgccac
//
