LOCUS       sequence-340208-        7332 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 7332)
  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 7332)
  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-CTSL
FEATURES             Location/Qualifiers
     source          1..7332
                     /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(4020..4044)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4091..4114
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4151..4174
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4205..4231
                     /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             4205..4231
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(4232..4579,4580..4582,4583..5191)
                     /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(5235..5252)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5241..5465
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5598..5617)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5749..5796
                     /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             5804..6826
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             5804..6826
                     /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    6956..7077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6993..7012)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7047..7066
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7126..7142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7126..7142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7139..7161)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7150..7166
                     /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(7174..7180,7181..7198,7199..7204))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7219..7240
                     /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 tccaccatga atcctacact catccttgct gccttttgcc tgggaattgc
     3061 ctcagctact ctaacatttg atcacagttt agaggcacag tggaccaagt ggaaggcgat
     3121 gcacaacaga ttatacggca tgaatgaaga aggatggagg agagcagtgt gggagaagaa
     3181 cgtgaagatg attgaactgc acaatcagga atacagggaa gggaaacaca gcttcacaat
     3241 ggccatgaac gcctttggag acatgaccag tgaagaattc aggcaggtga tgaatggctt
     3301 tcaaaaccgt aagcccagga aggggaaagt gttccaggaa cctctgtttt atgaggcccc
     3361 cagatctgtg gattggagag agaaaggcta cgtgactcct gtgaagaatc agggtcagtg
     3421 tggttcttgt tgggctttta gtgctactgg tgctcttgaa ggacagatgt tccggaaaac
     3481 tgggaggctt atctcactga gtgagcagaa tctggtagac tgctctgggc ctcaaggcaa
     3541 tgaaggctgc aatggtggcc taatggatta tgctttccag tatgttcagg ataatggagg
     3601 cctggactct gaggaatcct atccatatga ggcaacagaa gaatcctgta agtacaatcc
     3661 caagtattct gttgctaatg acaccggctt tgtggacatc cctaagcagg agaaggccct
     3721 gatgaaggca gttgcaactg tggggcccat ttctgttgct attgatgcag gtcatgagtc
     3781 cttcctgttc tataaagaag gcatttattt tgagccagac tgtagcagtg aagacatgga
     3841 tcatggtgtg ctggtggttg gctacggatt tgaaagcaca gaatcagata acaataaata
     3901 ttggctggtg aagaacagct ggggtgaaga atggggcatg ggtggctacg taaagatggc
     3961 caaagaccgg agaaaccatt gtggaattgc ctcagcagcc agctacccca ctgtgttgga
     4021 cccagctttc ttgtacaaag tggttgatat caaggccgcc gacatcacca gcctgtacaa
     4081 gaaagtgggg tggagccatc cccagttcga gaagggcgga ggatctggcg gcggaagcgg
     4141 cggaggaagc tggtcccacc ctcagtttga gaaaggcacc gagctgggca gcaccatggc
     4201 cagctacccc tacgacgtgc ccgactacgc caaggacaac accgtgcccc tgaagctgat
     4261 cgccctgctg gccaacggcg agttccactc tggcgagcag ctgggcgaga cactgggcat
     4321 gagcagagcc gccatcaaca agcacatcca gaccctgcgg gactggggcg tggacgtgtt
     4381 cacagtgccc ggcaagggct acagcctgcc cgagcccatc cagctgctga acgccaagca
     4441 gatcctgggc cagctggatg gcggatctgt ggccgtgctg cccgtgatcg acagcaccaa
     4501 ccagtatctg ctcgaccgga tcggcgagct gaagtccggc gacgcctgta tcgccgagta
     4561 ccagcaggcc ggcagaggcg gcaggggcag aaagtggttc agccccttcg gcgccaacct
     4621 gtacctgagc atgttctgga gactggaaca gggcccagcc gccgctattg gcctgagcct
     4681 ggtcatcggc atcgtgatgg ccgaggtgct gcggaagctg ggcgccgaca aagtgcgcgt
     4741 gaagtggccc aacgacctgt atctgcagga ccggaagctg gccggcatcc tggtggaact
     4801 gaccggcaaa acaggcgacg ccgcccagat cgtgatcggc gctggcatca acatggccat
     4861 gcggcgggtg gaagagagcg tggtcaacca gggctggatc accctgcagg aagccggcat
     4921 caacctggac cggaacaccc tggccgccat gctgatcaga gagctgagag ccgccctgga
     4981 actgttcgag caggaaggcc tggcccccta cctgagcaga tgggagaagc tggacaactt
     5041 catcaaccgg cccgtgaaac tgatcatcgg cgacaaagag atcttcggca tcagccgggg
     5101 catcgacaag cagggggccc tgctcctgga acaggacggc atcatcaagc cctggatggg
     5161 cggcgagatc agcctgcgga gcgccgagaa gtgatgactc gagtctagag ggcccgttta
     5221 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     5281 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     5341 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     5401 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     5461 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     5521 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5581 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5641 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5701 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtacctaga agttcctatt
     5761 ccgaagttcc tattctctag aaagtatagg aacttccttg gccaaaaagc ctgaactcac
     5821 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     5881 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     5941 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     6001 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct
     6061 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     6121 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
     6181 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     6241 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     6301 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     6361 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     6421 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     6481 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     6541 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct
     6601 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     6661 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     6721 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     6781 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tactacgaga
     6841 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6901 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     6961 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     7021 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     7081 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     7141 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7201 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7261 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7321 gagaggcggt tt
//
