LOCUS       sequence-364148-        6512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 ORF8 to TurboID and 3xHA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6512)
  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 6512)
  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: ORF8-1x-TurboID pbabe-puro (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..6512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             52..521
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    586..785
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             986..1402
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1332..1354
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1370..1386
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1808..2764
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             2771..2797
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2801..2827
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2831..2857
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2886..2906)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2891..3220
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3071..3206
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3230..3829
                     /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(3230..3249)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3726..3746
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4050..4519
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4649..4666)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4820..5408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4900..4919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5568..6359,6360..6428))
                     /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"
     promoter        complement(join(6429..6512,1..21))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgcag cctgaatatg
       61 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
      121 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
      181 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
      241 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
      301 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
      361 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
      421 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
      481 ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga
      541 tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggcaagct ggccagcaac
      601 ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta
      661 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac
      721 acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac
      781 ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga
      841 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag
      901 ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt
      961 gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt
     1021 aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag
     1081 acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga
     1141 cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc
     1201 gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc
     1261 ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc
     1321 cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc
     1381 cctcactcct tctctaggcg ccggcctcga ggccaccatg aagttcctgg tgttcctggg
     1441 tatcatcacc accgtggccg ctttccacca ggagtgctct ctgcagagct gcacccagca
     1501 ccagccatac gtggtggatg acccatgccc tatccacttc tacagcaagt ggtacatcag
     1561 agtgggagct aggaagtccg ctcctctgat cgagctgtgc gtggacgaag ccggcagcaa
     1621 gtccccaatc cagtacatcg acatcggaaa ctacaccgtg tcttgcctgc cattcaccat
     1681 caactgccag gaacccaagc tgggctccct ggtggtgaga tgctctttct acgaggactt
     1741 cctggaatac cacgacgtgc gtgtggtgct ggacttcatc accggtggtg gaggcgggtc
     1801 tgaattcaaa gacaatactg tgcctctgaa gctgatcgct ctcctggcta atggcgagtt
     1861 ccatagtggc gaacagctgg gagaaaccct gggcatgtcc agggccgcta tcaacaagca
     1921 cattcagact ctgcgcgact ggggcgtgga cgtgttcacc gtgcccggaa agggctactc
     1981 tctgcccgag cctatcccgc tgctgaacgc taaacagatt ctgggacagc tggacggcgg
     2041 gagcgtggca gtcctgcctg tggtcgactc caccaatcag tacctgctgg atcgaatcgg
     2101 cgagctgaag agtggggatg cttgcattgc agaatatcag caggcaggga gaggaagcag
     2161 agggaggaaa tggttctctc cttttggagc taacctgtac ctgagtatgt tttggcgcct
     2221 gaagcgggga ccagcagcaa tcggcctggg cccggtcatc ggaattgtca tggcagaagc
     2281 gctgcgaaag ctgggagcag acaaggtgcg agtcaaatgg cccaatgacc tgtatctgca
     2341 ggatagaaag ctggcaggca tcctggtgga gctggccgga ataacaggcg atgctgcaca
     2401 gatcgtcatt ggcgccggga ttaacgtggc tatgaggcgc gtggaggaaa gcgtggtcaa
     2461 tcagggctgg atcacactgc aggaagcagg gattaacctg gacaggaata ctctggccgc
     2521 tacgctgatc cgagagctgc gggcagccct ggaactgttc gagcaggaag gcctggctcc
     2581 atatctgcca cggtgggaga agctggataa cttcatcaat agacccgtga agctgatcat
     2641 tggggacaaa gagattttcg ggattagccg ggggattgat aaacagggag ccctgctgct
     2701 ggaacaggac ggagttatca aaccctggat gggcggagaa atcagtctgc ggtctgccga
     2761 aaagggatcc tacccgtatg atgttccgga ttacgctggc tatccctacg acgtgcccga
     2821 ctatgccggg tacccctatg acgtcccaga ctacgcatag gtttaaaccg ctgagtcgac
     2881 cctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2941 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3001 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3061 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3121 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3181 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttacca tgaccgagta
     3241 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     3301 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     3361 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     3421 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     3481 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     3541 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     3601 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     3661 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     3721 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     3781 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca
     3841 cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcgataa aataaaagat
     3901 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     3961 agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt
     4021 cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     4081 aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa
     4141 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
     4201 gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
     4261 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     4321 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc
     4381 cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
     4441 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
     4501 cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt aatttggttt tttttcttaa
     4561 gtatttacat taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4621 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4681 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4741 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4801 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4861 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4921 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4981 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5041 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5101 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5161 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5221 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5281 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5341 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5401 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5461 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttgc
     5521 ggccgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5581 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5641 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5701 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5761 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5821 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5881 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5941 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6001 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6061 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6121 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6181 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6241 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6301 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6361 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6421 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6481 agcggataca tatttgaatg tatttagaaa aa
//
