LOCUS       sequence-364139-        6034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 Envelope protein fused to BioID2 and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6034)
  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 6034)
  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: E-1x-BioID2 pBabe-puro (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..6034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             60..529
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    594..793
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             994..1410
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1340..1362
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1378..1394
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1679..2374
                     /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             2375..2401
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2416..2436)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2421..2750
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2601..2736
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2760..3359
                     /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(2760..2779)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3256..3276
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3580..4049
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4179..4196)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4350..4938)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4430..4449)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5098..5889,5890..5958))
                     /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     5721..5740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5959..6034,1..29))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgcagcc
       61 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
      121 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
      181 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
      241 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
      301 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
      361 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
      421 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
      481 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc
      541 gtccgggatc gggagacccc tgcccaggga ccaccgaccc accaccggga ggcaagctgg
      601 ccagcaactt atctgtgtct gtccgattgt ctagtgtcta tgactgattt tatgcgcctg
      661 cgtcggtact agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag
      721 ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt
      781 ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg
      841 gtaggagacg agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg
      901 aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt
      961 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
     1021 ttgaccttag gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc
     1081 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     1141 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     1201 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     1261 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct
     1321 ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt
     1381 tatccagccc tcactccttc tctaggcgcc ggccggatcc gccaccatgt actctttcgt
     1441 gagcgaggaa accggcaccc tgatcgtgaa ctccgtgctg ctgttcctgg ccttcgtggt
     1501 gttcctgctg gtgaccctgg ctatcctgac cgctctgaga ctgtgcgctt actgctgcaa
     1561 catcgtgaac gtgtccctgg tgaagccctc tttctacgtg tacagccgcg tgaagaacct
     1621 gaacagctcc agggtgcctg acctgctggt gaccggtggt ggaggcgggt ctgaattctt
     1681 caagaacctg atctggctga aggaggtgga cagcacccag gagagactga aggagtggaa
     1741 cgtgagctac ggcaccgccc tggtggccga cagacagacc aagggcagag gcggcctggg
     1801 cagaaagtgg ctgagccagg agggcggcct gtacttcagc ttcctgctga accccaagga
     1861 gttcgagaac ctgctgcagc tgcccctggt gctgggcctg agcgtgagcg aggccctgga
     1921 ggagatcacc gagatcccct tcagcctgaa gtggcccaac gacgtgtact tccaggagaa
     1981 gaaggtgagc ggcgtgctgt gcgagctgag caaggacaag ctgatcgtgg gcatcggcat
     2041 caacgtgaac cagagagaga tccccgagga gatcaaggac agagccacca ccctgtacga
     2101 gatcaccggc aaggactggg acagaaagga ggtgctgctg aaggtgctga agagaatcag
     2161 cgagaacctg aagaagttca aggagaagag cttcaaggag ttcaagggca agatcgagag
     2221 caagatgctg tacctgggcg aggaggtgaa gctgctgggc gagggcaaga tcaccggcaa
     2281 gctggtgggc ctgagcgaga agggcggcgc cctgatcctg accgaggagg gcatcaagga
     2341 gatcctgagc ggcgagttca gcctgagaag aagctaccct tacgatgtac cggattacgc
     2401 atgagtcgac cctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2461 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2521 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2581 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2641 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     2701 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttacca
     2761 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     2821 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     2881 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     2941 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     3001 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     3061 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     3121 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     3181 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     3241 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     3301 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac
     3361 gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcgataa
     3421 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     3481 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa
     3541 tagagaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga
     3601 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat
     3661 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     3721 atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag
     3781 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     3841 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     3901 ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc
     3961 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     4021 attgactacc cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt aatttggttt
     4081 tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg
     4141 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4201 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4261 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4321 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4381 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4441 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4501 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4561 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4621 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4681 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4741 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4801 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4861 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4921 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4981 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5041 atccttttgc ggccgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5101 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5161 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5221 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5281 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5341 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5401 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5461 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5521 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5581 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5641 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5701 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5761 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5821 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5881 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5941 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6001 ttgtctcatg agcggataca tatttgaatg tatt
//
