LOCUS       sequence-340223-        7623 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 7623)
  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 7623)
  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-TMPRSS4
FEATURES             Location/Qualifiers
     source          1..7623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        157..536
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        537..740
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     691..711
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    742..760
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    763..781
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     791..815
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(983..1333,1334..1336,1337..1945)
                     /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             1946..1972
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2003..2026
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2063..2086
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    2099..2123
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3008..3019)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(3435..3459)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3497..3514)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3503..3727
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3860..3879)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4011..4058
                     /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             4066..5088
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4066..5088
                     /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    5218..5339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5255..5274)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5309..5328
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5388..5404)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5388..5404)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5401..5423)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5412..5428
                     /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(5436..5442,5443..5460,5461..5466))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5481..5502
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5619..5636)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5790..6378)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5870..5889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6549..7340,7341..7409))
                     /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     7172..7191
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7410..7514)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7589..7608)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
       61 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
      121 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
      181 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      241 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      301 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      361 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      421 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      481 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      541 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      601 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      661 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      721 aggtctatat aagcagagct ctccctatca gtgatagaga tctccctatc agtgatagag
      781 atcgtcgacg agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
      841 ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagcg tttaaactta
      901 agcttggtac cgagctcgga tccactagtc cagtgtggtg gaattctgca gatatccagc
      961 acagtggcgg ccgctcgaga ccatgaagga caacaccgtg cccctgaagc tgatcgccct
     1021 gctggccaac ggcgagttcc actctggcga gcagctgggc gagacactgg gcatgagcag
     1081 agccgccatc aacaagcaca tccagaccct gcgggactgg ggcgtggacg tgttcacagt
     1141 gcccggcaag ggctacagcc tgcccgagcc catccagctg ctgaacgcca agcagatcct
     1201 gggccagctg gatggcggat ctgtggccgt gctgcccgtg atcgacagca ccaaccagta
     1261 tctgctcgac cggatcggcg agctgaagtc cggcgacgcc tgtatcgccg agtaccagca
     1321 ggccggcaga ggcggcaggg gcagaaagtg gttcagcccc ttcggcgcca acctgtacct
     1381 gagcatgttc tggagactgg aacagggccc agccgccgct attggcctga gcctggtcat
     1441 cggcatcgtg atggccgagg tgctgcggaa gctgggcgcc gacaaagtgc gcgtgaagtg
     1501 gcccaacgac ctgtatctgc aggaccggaa gctggccggc atcctggtgg aactgaccgg
     1561 caaaacaggc gacgccgccc agatcgtgat cggcgctggc atcaacatgg ccatgcggcg
     1621 ggtggaagag agcgtggtca accagggctg gatcaccctg caggaagccg gcatcaacct
     1681 ggaccggaac accctggccg ccatgctgat cagagagctg agagccgccc tggaactgtt
     1741 cgagcaggaa ggcctggccc cctacctgag cagatgggag aagctggaca acttcatcaa
     1801 ccggcccgtg aaactgatca tcggcgacaa agagatcttc ggcatcagcc ggggcatcga
     1861 caagcagggg gccctgctcc tggaacagga cggcatcatc aagccctgga tgggcggcga
     1921 gatcagcctg cggagcgccg agaagtaccc atacgatgtt cctgactatg ccggtaccga
     1981 gctcggatcc accatggcta gctggagcca cccgcagttc gagaaaggtg gaggttccgg
     2041 aggtggatcg ggaggtggat cgtggagcca cccgcagttc gaaaaagcgg ccgatatcac
     2101 aagtttgtac aaaaaagttg gcatgttaca ggatcctgac agtgatcaac ctctgaacag
     2161 cctcgatgtc aaacccctgc gcaaaccccg tatccccatg gagaccttca gaaaggtggg
     2221 gatccccatc atcatagcac tactgagcct ggcgagtatc atcattgtgg ttgtcctcat
     2281 caaggtgatt ctggataaat actacttcct ctgcgggcag cctctccact tcatcccgag
     2341 gaagcagctg tgtgacggag agctggactg tcccttgggg gaggacgagg agcactgtgt
     2401 caagagcttc cccgaagggc ctgcagtggc agtccgcctc tccaaggacc gatccacact
     2461 gcaggtgctg gactcggcca cagggaactg gttctctgcc tgtttcgaca acttcacaga
     2521 agctctcgct gagacagcct gtaggcagat gggctacagc agcaaaccca ctttcagagc
     2581 tgtggagatt ggcccagacc aggatctgga tgttgttgaa atcacagaaa acagccagga
     2641 gcttcgcatg cggaactcaa gtgggccctg tctctcaggc tccctggtct ccctgcactg
     2701 tcttgcctgt gggaagagcc tgaagacccc ccgtgtggtg ggtggggagg aggcctctgt
     2761 ggattcttgg ccttggcagg tcagcatcca gtacgacaaa cagcacgtct gtggagggag
     2821 catcctggac ccccactggg tcctcacggc agcccactgc ttcaggaaac ataccgatgt
     2881 gttcaactgg aaggtgcggg caggctcaga caaactgggc agcttcccat ccctggctgt
     2941 ggccaagatc atcatcattg aattcaaccc catgtacccc aaagacaatg acatcgccct
     3001 catgaagctg cagttcccac tcactttctc aggcacagtc aggcccatct gtctgccctt
     3061 ctttgatgag gagctcactc cagccacccc actctggatc attggatggg gctttacgaa
     3121 gcagaatgga gggaagatgt ctgacatact gctgcaggcg tcagtccagg tcattgacag
     3181 cacacggtgc aatgcagacg atgcgtacca gggggaagtc accgagaaga tgatgtgtgc
     3241 aggcatcccg gaagggggtg tggacacctg ccagggtgac agtggtgggc ccctgatgta
     3301 ccaatctgac cagtggcatg tggtgggcat cgttagctgg ggctatggct gcgggggccc
     3361 gagcacccca ggagtataca ccaaggtctc agcctatctc aactggatct acaatgtctg
     3421 gaaggctgag ctgtacccaa ctttcttgta caaagtggtg acgtaagcta ggggcccgtt
     3481 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3541 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3601 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3661 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3721 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3781 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3841 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3901 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3961 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta gaagttccta
     4021 ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa gcctgaactc
     4081 accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg
     4141 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat
     4201 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac
     4261 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt cagcgagagc
     4321 ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc
     4381 gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat
     4441 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca
     4501 tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg
     4561 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag
     4621 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg
     4681 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa
     4741 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg
     4801 cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg
     4861 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca
     4921 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt
     4981 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc
     5041 gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca cgtactacga
     5101 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5161 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5221 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5281 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5341 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5401 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5461 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5521 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5581 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5641 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5701 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5761 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5821 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5881 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5941 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6001 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6061 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6121 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6181 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6241 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6301 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6361 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6421 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6481 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6541 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6601 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6661 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6721 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6781 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6841 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6901 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6961 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7021 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7081 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7141 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7201 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7261 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7321 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7381 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7441 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7501 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     7561 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     7621 agt
//
