LOCUS       sequence-340213-        6696 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 6696)
  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 6696)
  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-IFITM1
FEATURES             Location/Qualifiers
     source          1..6696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        143..522
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        523..726
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     677..697
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    728..746
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    749..767
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     777..801
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(969..1319,1320..1322,1323..1931)
                     /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             1932..1958
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1989..2012
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2049..2072
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    2085..2109
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2494..2518)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2556..2573)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2562..2786
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2919..2938)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3070..3117
                     /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             3125..4147
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3125..4147
                     /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    4277..4398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4314..4333)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4368..4387
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4447..4463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4447..4463)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4460..4482)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4471..4487
                     /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(4495..4501,4502..4519,4520..4525))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4540..4561
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4678..4695)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4849..5437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4929..4948)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5608..6399,6400..6468))
                     /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     6231..6250
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6469..6573)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6648..6667)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
       61 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      121 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      181 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      241 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      301 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      361 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      421 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      481 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      541 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      601 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      661 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      721 agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg tcgacgagct
      781 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
      841 agacaccggg accgatccag cctccggact ctagcgttta aacttaagct tggtaccgag
      901 ctcggatcca ctagtccagt gtggtggaat tctgcagata tccagcacag tggcggccgc
      961 tcgagaccat gaaggacaac accgtgcccc tgaagctgat cgccctgctg gccaacggcg
     1021 agttccactc tggcgagcag ctgggcgaga cactgggcat gagcagagcc gccatcaaca
     1081 agcacatcca gaccctgcgg gactggggcg tggacgtgtt cacagtgccc ggcaagggct
     1141 acagcctgcc cgagcccatc cagctgctga acgccaagca gatcctgggc cagctggatg
     1201 gcggatctgt ggccgtgctg cccgtgatcg acagcaccaa ccagtatctg ctcgaccgga
     1261 tcggcgagct gaagtccggc gacgcctgta tcgccgagta ccagcaggcc ggcagaggcg
     1321 gcaggggcag aaagtggttc agccccttcg gcgccaacct gtacctgagc atgttctgga
     1381 gactggaaca gggcccagcc gccgctattg gcctgagcct ggtcatcggc atcgtgatgg
     1441 ccgaggtgct gcggaagctg ggcgccgaca aagtgcgcgt gaagtggccc aacgacctgt
     1501 atctgcagga ccggaagctg gccggcatcc tggtggaact gaccggcaaa acaggcgacg
     1561 ccgcccagat cgtgatcggc gctggcatca acatggccat gcggcgggtg gaagagagcg
     1621 tggtcaacca gggctggatc accctgcagg aagccggcat caacctggac cggaacaccc
     1681 tggccgccat gctgatcaga gagctgagag ccgccctgga actgttcgag caggaaggcc
     1741 tggcccccta cctgagcaga tgggagaagc tggacaactt catcaaccgg cccgtgaaac
     1801 tgatcatcgg cgacaaagag atcttcggca tcagccgggg catcgacaag cagggggccc
     1861 tgctcctgga acaggacggc atcatcaagc cctggatggg cggcgagatc agcctgcgga
     1921 gcgccgagaa gtacccatac gatgttcctg actatgccgg taccgagctc ggatccacca
     1981 tggctagctg gagccacccg cagttcgaga aaggtggagg ttccggaggt ggatcgggag
     2041 gtggatcgtg gagccacccg cagttcgaaa aagcggccga tatcacaagt ttgtacaaaa
     2101 aagcaggctc caccatgcac aaggaggaac atgaggtggc tgtgctgggg gcacccccca
     2161 gcaccatcct tccaaggtcc accgtgatca acatccacag cgagacctcc gtgcccgacc
     2221 atgtcgtctg gtccctgttc aacaccctct tcttgaactg gtgctgtctg ggcttcatag
     2281 cattcgccta ctccgtgaag tctagggaca ggaagatggt tggcgacgtg accggggccc
     2341 aggcctatgc ctccaccgcc aagtgcctga acatctgggc cctgattctg ggcatcctca
     2401 tgaccattgg attcatcctg ttactggtat tcggctctgt gacagtctac catattatgt
     2461 tacagataat acaggaaaaa cggggttact aggacccagc tttcttgtac aaagtggtga
     2521 cgtaagctag gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     2581 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2641 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2701 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2761 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     2821 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2881 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2941 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3001 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3061 acgtacctag aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcctt
     3121 ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     3181 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     3241 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     3301 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
     3361 tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     3421 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga
     3481 tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     3541 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     3601 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     3661 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     3721 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     3781 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     3841 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     3901 gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     3961 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     4021 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg
     4081 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     4141 ggaatagcac gtactacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4201 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4261 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4321 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4381 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     4441 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4501 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4561 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4621 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4681 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4741 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4801 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4861 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4921 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4981 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5041 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5101 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5161 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5221 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5281 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5341 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5401 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5461 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5521 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5581 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5641 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5701 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5761 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5821 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5881 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5941 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6001 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6061 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6121 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6181 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6241 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6301 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6361 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6421 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6481 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6541 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6601 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     6661 gatgccgcat agttaagcca gtatctgctc cctgct
//
