LOCUS       sequence-364147-        6003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BioID2 fused to SARS-CoV-2 ORF7b and myc tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6003)
  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 6003)
  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: BioID2-1x-ORF7b pBabe-puro (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..6003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             111..580
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    645..844
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1045..1461
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1391..1413
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1429..1445
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1514..1523
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1523..1552
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1556..2251
                     /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"
     primer_bind     complement(2436..2456)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2441..2770
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2621..2756
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2780..3379
                     /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(2780..2799)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3276..3296
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3600..4069
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4199..4216)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4370..4958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4450..4469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5118..5909,5910..5978))
                     /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     5741..5760
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5979..6003,1..80))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
       61 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgcagc ctgaatatgg
      121 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
      181 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
      241 gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca
      301 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
      361 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
      421 acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta
      481 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
      541 tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct cgtccgggat
      601 cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg gccagcaact
      661 tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct gcgtcggtac
      721 tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca
      781 cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc
      841 tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct ggtaggagac
      901 gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg gaaccgaagc
      961 cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg tctgactgtg
     1021 tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag tttgacctta
     1081 ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt caagaagaga
     1141 cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg gccgcgagac
     1201 ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc acctggcccg
     1261 catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc ttttgacccc
     1321 cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc tccatccgcc
     1381 ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct ttatccagcc
     1441 ctcactcctt ctctaggcgc cggccggatc ccagtgtggt ggtacgtagg aattcggaga
     1501 cccaagctgg ctagccacca tggaacaaaa actcatctca gaagaggatc tcgacttcaa
     1561 gaacctgatc tggctgaagg aggtggacag cacccaggag agactgaagg agtggaacgt
     1621 gagctacggc accgccctgg tggccgacag acagaccaag ggcagaggcg gcctgggcag
     1681 aaagtggctg agccaggagg gcggcctgta cttcagcttc ctgctgaacc ccaaggagtt
     1741 cgagaacctg ctgcagctgc ccctggtgct gggcctgagc gtgagcgagg ccctggagga
     1801 gatcaccgag atccccttca gcctgaagtg gcccaacgac gtgtacttcc aggagaagaa
     1861 ggtgagcggc gtgctgtgcg agctgagcaa ggacaagctg atcgtgggca tcggcatcaa
     1921 cgtgaaccag agagagatcc ccgaggagat caaggacaga gccaccaccc tgtacgagat
     1981 caccggcaag gactgggaca gaaaggaggt gctgctgaag gtgctgaaga gaatcagcga
     2041 gaacctgaag aagttcaagg agaagagctt caaggagttc aagggcaaga tcgagagcaa
     2101 gatgctgtac ctgggcgagg aggtgaagct gctgggcgag ggcaagatca ccggcaagct
     2161 ggtgggcctg agcgagaagg gcggcgccct gatcctgacc gaggagggca tcaaggagat
     2221 cctgagcggc gagttcagcc tgagaagaag cctcgagggt ggaggcgggt ctaccggtat
     2281 gatcgagctg agcctgatcg acttctacct gtgcttcctg gccttcctgc tgttcctggt
     2341 gctgatcatg ctgatcatct tctggttctc cctggagctg caggaccaca acgaaacctg
     2401 ccacgcttaa gtttaaaccg ctgagtcgac cctgtggaat gtgtgtcagt tagggtgtgg
     2461 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2521 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2581 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2641 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2701 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2761 cttttgcaaa aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     2821 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     2881 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     2941 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     3001 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     3061 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     3121 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     3181 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     3241 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     3301 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     3361 ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc
     3421 acgaccccat gcatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     3481 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     3541 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     3601 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     3661 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     3721 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     3781 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     3841 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     3901 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     3961 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     4021 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg
     4081 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt
     4141 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4201 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4261 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4321 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4381 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4441 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4501 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4561 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4621 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4681 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4741 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4801 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4861 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4921 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4981 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5041 caaaaaggat cttcacctag atccttttgc ggccgcaaat caatctaaag tatatatgag
     5101 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5161 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5221 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5281 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5341 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5401 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5461 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5521 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5581 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5641 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5701 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5761 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5821 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5881 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5941 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6001 tga
//
