LOCUS       Exported                6229 bp ds-DNA     circular SYN 16-JUN-2022
DEFINITION  Expresses SARS-CoV-2 NSP10 fused to BioID2 and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    NSP10-1x-BioID2 pbabe-puro (SARS-CoV-2)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6229)
  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 6229)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 16, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6229
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     regulatory      1369..1378
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1822..2517
                     /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             2518..2544
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2559..2579)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2564..2893
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2744..2879
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2806..2825
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2903..3502
                     /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(2903..2922)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3399..3419
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3723..4192
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4322..4339)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4493..5081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4573..4592)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5241..6101)
                     /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     5864..5883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6102..6206)
                     /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 caccatggct
     1381 ggtaacgcta ctgaggtgcc tgctaacagc accgtgctgt ccttctgcgc cttcgctgtg
     1441 gacgccgcta aggcctacaa ggactacctg gcttccggcg gacagcctat caccaactgc
     1501 gtgaagatgc tgtgcaccca caccggaacc ggtcaggcca tcaccgtgac cccagaggct
     1561 aacatggacc aggaatcttt cggtggcgcc agctgctgcc tgtactgcag atgccacatc
     1621 gaccacccca accctaaggg tttctgcgac ctgaagggca agtacgtgca gatcccaacc
     1681 acctgcgcta acgaccccgt gggattcacc ctgaagaaca ccgtgtgcac cgtgtgcgga
     1741 atgtggaagg gctacggatg ctcttgcgac cagctgaggg aacctatgct gcagaccggt
     1801 ggtggaggcg ggtctgaatt cttcaagaac ctgatctggc tgaaggaggt ggacagcacc
     1861 caggagagac tgaaggagtg gaacgtgagc tacggcaccg ccctggtggc cgacagacag
     1921 accaagggca gaggcggcct gggcagaaag tggctgagcc aggagggcgg cctgtacttc
     1981 agcttcctgc tgaaccccaa ggagttcgag aacctgctgc agctgcccct ggtgctgggc
     2041 ctgagcgtga gcgaggccct ggaggagatc accgagatcc ccttcagcct gaagtggccc
     2101 aacgacgtgt acttccagga gaagaaggtg agcggcgtgc tgtgcgagct gagcaaggac
     2161 aagctgatcg tgggcatcgg catcaacgtg aaccagagag agatccccga ggagatcaag
     2221 gacagagcca ccaccctgta cgagatcacc ggcaaggact gggacagaaa ggaggtgctg
     2281 ctgaaggtgc tgaagagaat cagcgagaac ctgaagaagt tcaaggagaa gagcttcaag
     2341 gagttcaagg gcaagatcga gagcaagatg ctgtacctgg gcgaggaggt gaagctgctg
     2401 ggcgagggca agatcaccgg caagctggtg ggcctgagcg agaagggcgg cgccctgatc
     2461 ctgaccgagg agggcatcaa ggagatcctg agcggcgagt tcagcctgag aagaagctac
     2521 ccttacgatg taccggatta cgcatgagtc gaccctgtgg aatgtgtgtc agttagggtg
     2581 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2641 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2701 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2761 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2821 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2881 aggcttttgc aaaaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     2941 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3001 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3061 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     3121 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     3181 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     3241 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     3301 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     3361 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     3421 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     3481 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     3541 cgcacgaccc catgcatcga taaaataaaa gattttattt agtctccaga aaaagggggg
     3601 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     3661 atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac
     3721 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     3781 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     3841 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
     3901 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     3961 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     4021 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
     4081 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     4141 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc
     4201 atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc
     4261 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4321 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4381 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4441 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4501 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4561 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4621 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4681 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4741 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4801 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4861 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4921 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4981 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5041 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5101 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5161 tatcaaaaag gatcttcacc tagatccttt tgcggccgca aatcaatcta aagtatatat
     5221 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5281 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5341 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5401 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5461 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5521 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5581 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5641 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5701 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5761 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5821 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5881 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5941 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6001 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6061 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6121 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6181 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccac
//
