LOCUS       sequence-364144-        5992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SARS-CoV-2 ORF6 fused to BioID2 and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5992)
  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 5992)
  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: ORF6-1x-BioID2 pBabe-puro (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..5992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             56..525
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    590..789
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             990..1406
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1336..1358
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1374..1390
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1633..2328
                     /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             2329..2355
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2370..2390)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2375..2704
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2555..2690
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2714..3313
                     /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(2714..2733)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3210..3230
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3534..4003
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4133..4150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4304..4892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4384..4403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5052..5843,5844..5912))
                     /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     5675..5694
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5913..5992,1..25))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gcagcctgaa
       61 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
      121 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
      181 tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac
      241 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
      301 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
      361 agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc
      421 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
      481 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc
      541 gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggca agctggccag
      601 caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc
      661 ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct
      721 gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc
      781 cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag
      841 gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc
      901 gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga
      961 ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga
     1021 ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga
     1081 agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc
     1141 gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg
     1201 gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg
     1261 acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat
     1321 ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc
     1381 cagccctcac tccttctcta ggcgccggcc ggatccgcca ccatgttcca cctggtggac
     1441 ttccaggtga ccatcgccga gatcctgctg atcatcatgc gcaccttcaa ggtgtctatc
     1501 tggaacctgg actacatcat caacctgatc atcaagaacc tgagcaagtc cctgaccgag
     1561 aacaagtaca gccagctgga cgaggaacag cccatggaaa tcgacaccgg tggtggaggc
     1621 gggtctgaat tcttcaagaa cctgatctgg ctgaaggagg tggacagcac ccaggagaga
     1681 ctgaaggagt ggaacgtgag ctacggcacc gccctggtgg ccgacagaca gaccaagggc
     1741 agaggcggcc tgggcagaaa gtggctgagc caggagggcg gcctgtactt cagcttcctg
     1801 ctgaacccca aggagttcga gaacctgctg cagctgcccc tggtgctggg cctgagcgtg
     1861 agcgaggccc tggaggagat caccgagatc cccttcagcc tgaagtggcc caacgacgtg
     1921 tacttccagg agaagaaggt gagcggcgtg ctgtgcgagc tgagcaagga caagctgatc
     1981 gtgggcatcg gcatcaacgt gaaccagaga gagatccccg aggagatcaa ggacagagcc
     2041 accaccctgt acgagatcac cggcaaggac tgggacagaa aggaggtgct gctgaaggtg
     2101 ctgaagagaa tcagcgagaa cctgaagaag ttcaaggaga agagcttcaa ggagttcaag
     2161 ggcaagatcg agagcaagat gctgtacctg ggcgaggagg tgaagctgct gggcgagggc
     2221 aagatcaccg gcaagctggt gggcctgagc gagaagggcg gcgccctgat cctgaccgag
     2281 gagggcatca aggagatcct gagcggcgag ttcagcctga gaagaagcta cccttacgat
     2341 gtaccggatt acgcatgagt cgaccctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2401 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2461 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2521 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2581 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2641 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2701 caaaaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     2761 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     2821 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     2881 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     2941 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     3001 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     3061 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     3121 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     3181 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     3241 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     3301 gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc
     3361 ccatgcatcg ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     3421 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa
     3481 tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata
     3541 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     3601 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     3661 agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca
     3721 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
     3781 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     3841 cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt
     3901 gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg
     3961 gtctcctctg agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa
     4021 aattaatttg gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa
     4081 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4141 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4201 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4261 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4321 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4381 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4441 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4501 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4561 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4621 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4681 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4741 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4801 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4861 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4921 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4981 ggatcttcac ctagatcctt ttgcggccgc aaatcaatct aaagtatata tgagtaaact
     5041 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5101 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5161 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5221 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5281 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5341 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5401 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5461 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5521 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5581 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5641 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5701 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5761 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5821 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5881 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5941 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta ga
//
