LOCUS       sequence-340205-        6966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAC-tagged gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6966)
  AUTHORS   Liu X, Huuskonen S, Laitinen T, Redchuk T, Bogacheva M, Salokas K, 
            Pohner I, Ohman T, Tonduru AK, Hassinen A, Gawriyski L, Keskitalo S,
            Vartiainen MK, Pietiainen V, Poso A, Varjosalo M
  TITLE     SARS-CoV-2-host proteome interactions for antiviral drug discovery.
  JOURNAL   Mol Syst Biol. 2021 Nov;17(11):e10396. doi: 10.15252/msb.202110396.
  PUBMED    34709727
REFERENCE   2  (bases 1 to 6966)
  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: "Mol Syst 
            Biol."; date: "2021-11"; volume: "17(11):e10396. doi"; pages: " 
            10.15252/msb.202110396"
COMMENT     Sequence Label: MAC-CLEC4D
FEATURES             Location/Qualifiers
     source          1..6966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        134..513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        514..717
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     668..688
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    719..737
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    740..758
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     768..792
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(960..1310,1311..1313,1314..1922)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             1923..1949
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1980..2003
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2040..2063
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    2076..2100
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2755..2779)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2817..2834)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2823..3047
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3180..3199)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3331..3378
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3386..4408
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3386..4408
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4538..4659
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4575..4594)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4629..4648
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4708..4724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4708..4724)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4721..4743)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4732..4748
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4756..4762,4763..4780,4781..4786))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4801..4822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4939..4956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5110..5698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5190..5209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5869..6660,6661..6729))
                     /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     6492..6511
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6730..6834)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6909..6928)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
       61 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
      121 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      481 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      541 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      601 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      661 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
      721 cctatcagtg atagagatct ccctatcagt gatagagatc gtcgacgagc tcgtttagtg
      781 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
      841 gaccgatcca gcctccggac tctagcgttt aaacttaagc ttggtaccga gctcggatcc
      901 actagtccag tgtggtggaa ttctgcagat atccagcaca gtggcggccg ctcgagacca
      961 tgaaggacaa caccgtgccc ctgaagctga tcgccctgct ggccaacggc gagttccact
     1021 ctggcgagca gctgggcgag acactgggca tgagcagagc cgccatcaac aagcacatcc
     1081 agaccctgcg ggactggggc gtggacgtgt tcacagtgcc cggcaagggc tacagcctgc
     1141 ccgagcccat ccagctgctg aacgccaagc agatcctggg ccagctggat ggcggatctg
     1201 tggccgtgct gcccgtgatc gacagcacca accagtatct gctcgaccgg atcggcgagc
     1261 tgaagtccgg cgacgcctgt atcgccgagt accagcaggc cggcagaggc ggcaggggca
     1321 gaaagtggtt cagccccttc ggcgccaacc tgtacctgag catgttctgg agactggaac
     1381 agggcccagc cgccgctatt ggcctgagcc tggtcatcgg catcgtgatg gccgaggtgc
     1441 tgcggaagct gggcgccgac aaagtgcgcg tgaagtggcc caacgacctg tatctgcagg
     1501 accggaagct ggccggcatc ctggtggaac tgaccggcaa aacaggcgac gccgcccaga
     1561 tcgtgatcgg cgctggcatc aacatggcca tgcggcgggt ggaagagagc gtggtcaacc
     1621 agggctggat caccctgcag gaagccggca tcaacctgga ccggaacacc ctggccgcca
     1681 tgctgatcag agagctgaga gccgccctgg aactgttcga gcaggaaggc ctggccccct
     1741 acctgagcag atgggagaag ctggacaact tcatcaaccg gcccgtgaaa ctgatcatcg
     1801 gcgacaaaga gatcttcggc atcagccggg gcatcgacaa gcagggggcc ctgctcctgg
     1861 aacaggacgg catcatcaag ccctggatgg gcggcgagat cagcctgcgg agcgccgaga
     1921 agtacccata cgatgttcct gactatgccg gtaccgagct cggatccacc atggctagct
     1981 ggagccaccc gcagttcgag aaaggtggag gttccggagg tggatcggga ggtggatcgt
     2041 ggagccaccc gcagttcgaa aaagcggccg atatcacaag tttgtacaaa aaagcaggct
     2101 ccaccatggg gctagaaaaa cctcaaagta aactggaagg aggcatgcat ccccagctga
     2161 taccttcggt tattgctgta gttttcatct tacttctcag tgtctgtttt attgcaagtt
     2221 gtttggtgac tcatcacaac ttttcacgct gtaagagagg cacaggagtg cacaagttag
     2281 agcaccatgc aaagctcaaa tgcatcaaag agaaatcaga actgaaaagt gctgaaggga
     2341 gcacctggaa ctgttgtcct attgactgga gagccttcca gtccaactgc tattttcctc
     2401 ttactgacaa caagacgtgg gctgagagtg aaaggaactg ttcagggatg ggggcccatc
     2461 tgatgaccat cagcacggaa gctgagcaga actttattat tcagtttctg gatagacggc
     2521 tttcctattt ccttggactt agagatgaga atgccaaagg tcagtggcgt tgggtggacc
     2581 agacgccatt taacccacgc agagtattct ggcataagaa tgaacccgac aactctcagg
     2641 gagaaaactg tgttgttctt gtttataacc aagataaatg ggcctggaat gatgttcctt
     2701 gtaactttga agcaagtagg atttgtaaaa tacctggaac aacattgaac ttggacccag
     2761 ctttcttgta caaagtggtg acgtaagcta ggggcccgtt taaacccgct gatcagcctc
     2821 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2881 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2941 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3001 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3061 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3121 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3181 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3241 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3301 acttgattag ggtgatggtt cacgtaccta gaagttccta ttccgaagtt cctattctct
     3361 agaaagtata ggaacttcct tggccaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     3421 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     3481 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     3541 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc
     3601 gattccggaa gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg
     3661 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     3721 gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt
     3781 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     3841 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     3901 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     3961 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     4021 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     4081 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     4141 tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca
     4201 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     4261 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     4321 cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     4381 cagcactcgt ccgagggcaa aggaatagca cgtactacga gatttcgatt ccaccgccgc
     4441 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4501 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4561 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4621 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4681 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4741 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4801 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4861 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4921 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4981 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5041 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5101 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5161 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5221 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5281 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5341 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5401 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5461 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5521 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5581 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5641 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5701 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5761 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5821 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5881 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5941 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6001 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6061 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6121 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6181 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6241 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6301 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6361 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6421 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6481 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6541 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6601 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6661 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6721 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6781 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6841 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     6901 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     6961 gtgttg
//
