LOCUS       sequence-340227-        7347 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 7347)
  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 7347)
  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-CTSB
FEATURES             Location/Qualifiers
     source          1..7347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..42)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(196..784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(276..295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(955..1746,1747..1815))
                     /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     1578..1597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1816..1920)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1995..2014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2186..2565
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2566..2769
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2720..2740
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2771..2789
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2792..2810
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2820..2844
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2987..3011
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4035..4059)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4106..4129
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4166..4189
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4220..4246
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             4220..4246
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(4247..4594,4595..4597,4598..5206)
                     /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="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     complement(5250..5267)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5256..5480
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5613..5632)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5764..5811
                     /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             5819..6841
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             5819..6841
                     /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    6971..7092
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7008..7027)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7062..7081
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7141..7157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7141..7157)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7154..7176)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7165..7181
                     /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(7189..7195,7196..7213,7214..7219))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7234..7255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
       61 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      841 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1801 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1861 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1921 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     1981 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     2041 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     2101 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     2161 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     2221 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     2281 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     2341 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     2401 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     2461 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     2521 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     2581 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     2641 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     2701 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     2761 agcagagctc tccctatcag tgatagagat ctccctatca gtgatagaga tcgtcgacga
     2821 gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat
     2881 agaagacacc gggaccgatc cagcctccgg actctagcgt ttaaacttaa gcttggtacc
     2941 gagctcggat ccactagtcc agtgtggtgg aattctgcag atatcaacaa gtttgtacaa
     3001 aaaagcaggc tacaaaatgt ggcagctctg ggcctccctc tgctgcctgc tggtgttggc
     3061 caatgcccgg agcaggccct ctttccatcc cttaagcgat gagctggtca actatgtcaa
     3121 caaacggaat accacgtggc aggccgggca caacttctac aacgtggaca tgagctactt
     3181 gaagaggcta tgtggtacct tcctgggtgg gcccaagcca ccccagagag ttatgtttac
     3241 cgaggacctg aagctgcctg caagcttcga tgcacgggaa cgatggccac agtgtcccac
     3301 catcaaagag atcagagacc agggctcctg tggcagctgc tgggccttcg gggctgtgga
     3361 agccatctct gaccggatct gcatccacac caatgcgcac gtcagcgtgg aggtgtcggc
     3421 ggaggacctg ctcacctgct gtggcagcat gtgtggggac ggctgtaatg gtggctatcc
     3481 tgctgaagct tggaacttct ggacaagaaa aggcctggtt tctggtggcc tctatgaatc
     3541 ccatgtaggg tgcagaccgt actccatccc tccctgtgag caccacgtca acggctcccg
     3601 gcccccatgc acgggggagg gagatacccc caagtgtagc aagatctgtg agcctggcta
     3661 cagcccgacc tacaaacagg acaagcacta cggatacaat tcctacagcg tctccaatag
     3721 cgagaaggac atcatggccg agatctacaa aaacggcccc gtggagggag ctttctctgt
     3781 gtattcggac ttcctgctct acaagtcagg agtgtaccaa cacgtcaccg gagagatgat
     3841 gggtggccat gccatccgca tcctgggctg gggagtggag aatggcacac cctactggct
     3901 ggttgccaac tcctggaaca ctgactgggg tgacaatggc ttctttaaaa tactcagagg
     3961 acaggatcac tgtggaatcg aatcagaagt ggtggctgga attccacgca ccgatcagta
     4021 ctgggaaaag atcaacccag ctttcttgta caaagtggtt gatatcaagg ccgccgacat
     4081 caccagcctg tacaagaaag tggggtggag ccatccccag ttcgagaagg gcggaggatc
     4141 tggcggcgga agcggcggag gaagctggtc ccaccctcag tttgagaaag gcaccgagct
     4201 gggcagcacc atggccagct acccctacga cgtgcccgac tacgccaagg acaacaccgt
     4261 gcccctgaag ctgatcgccc tgctggccaa cggcgagttc cactctggcg agcagctggg
     4321 cgagacactg ggcatgagca gagccgccat caacaagcac atccagaccc tgcgggactg
     4381 gggcgtggac gtgttcacag tgcccggcaa gggctacagc ctgcccgagc ccatccagct
     4441 gctgaacgcc aagcagatcc tgggccagct ggatggcgga tctgtggccg tgctgcccgt
     4501 gatcgacagc accaaccagt atctgctcga ccggatcggc gagctgaagt ccggcgacgc
     4561 ctgtatcgcc gagtaccagc aggccggcag aggcggcagg ggcagaaagt ggttcagccc
     4621 cttcggcgcc aacctgtacc tgagcatgtt ctggagactg gaacagggcc cagccgccgc
     4681 tattggcctg agcctggtca tcggcatcgt gatggccgag gtgctgcgga agctgggcgc
     4741 cgacaaagtg cgcgtgaagt ggcccaacga cctgtatctg caggaccgga agctggccgg
     4801 catcctggtg gaactgaccg gcaaaacagg cgacgccgcc cagatcgtga tcggcgctgg
     4861 catcaacatg gccatgcggc gggtggaaga gagcgtggtc aaccagggct ggatcaccct
     4921 gcaggaagcc ggcatcaacc tggaccggaa caccctggcc gccatgctga tcagagagct
     4981 gagagccgcc ctggaactgt tcgagcagga aggcctggcc ccctacctga gcagatggga
     5041 gaagctggac aacttcatca accggcccgt gaaactgatc atcggcgaca aagagatctt
     5101 cggcatcagc cggggcatcg acaagcaggg ggccctgctc ctggaacagg acggcatcat
     5161 caagccctgg atgggcggcg agatcagcct gcggagcgcc gagaagtgat gactcgagtc
     5221 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     5281 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     5341 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     5401 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     5461 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     5521 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5581 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5641 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5701 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac
     5761 ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa
     5821 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     5881 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     5941 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     6001 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     6061 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     6121 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     6181 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     6241 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     6301 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     6361 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     6421 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     6481 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     6541 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     6601 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     6661 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     6721 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     6781 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     6841 gcacgtacta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6901 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6961 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     7021 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     7081 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     7141 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7201 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7261 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7321 cggccaacgc gcggggagag gcggttt
//
