LOCUS       sequence-384054-        7188 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Sars-CoV-2 NSP6_S107-109del in MAC-C destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7188)
  AUTHORS   Varjosalo M, Huuskonen S, Liu X, Redchuk T, Salokas K, Lundberg R, 
            Maljanen S, Belik M, Reinholm A, Kolehmainen P, Tripathi G, Laine P,
            Belanov S, Auvinen P, Vartiainen M, Laine L, Osterlund P, Julkkunen 
            I, Kakkola L
  TITLE     The comprehensive SARS-CoV-2 'hijackome' knowledge base— reveals 
            significant changes in host cell protein expression and activation 
            by multiple SARS-CoV-2 variants
  JOURNAL   10 May 2024, Research Square
REFERENCE   2  (bases 1 to 7188)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "10 May 
            2024, Research Square"
COMMENT     Sequence Label: MAC-C Sars-CoV-2 NSP6_S107-109del
FEATURES             Location/Qualifiers
     source          1..7188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(160..748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(240..259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(919..1710,1711..1779))
                     /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     1542..1561
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1780..1884)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1959..1978)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2150..2529
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2530..2733
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2684..2704
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2735..2753
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2756..2774
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2784..2808
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2951..2975
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3840..3864)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3911..3934
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3971..3994
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4025..4051
                     /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             4025..4051
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(4052..4399,4400..4402,4403..5011)
                     /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(5055..5072)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5061..5285
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5418..5437)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5569..5616
                     /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             5624..6646
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             5624..6646
                     /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    6776..6897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6813..6832)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6867..6886
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6946..6962)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6946..6962)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6959..6981)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6970..6986
                     /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(6994..7000,7001..7018,7019..7024))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7039..7060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(7177..7188,1..6))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
       61 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
      121 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      181 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      241 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      301 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      361 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      421 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
      481 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
      541 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
      601 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
      661 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
      721 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
      781 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
      841 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
      901 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
      961 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     1021 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     1081 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     1141 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     1201 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     1261 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     1321 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     1381 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     1441 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     1501 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     1561 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     1621 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     1681 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     1741 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     1801 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     1861 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     1921 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     1981 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     2041 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     2101 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
     2161 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     2221 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     2281 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     2341 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     2401 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     2461 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     2521 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     2581 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     2641 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     2701 cgtgtacggt gggaggtcta tataagcaga gctctcccta tcagtgatag agatctccct
     2761 atcagtgata gagatcgtcg acgagctcgt ttagtgaacc gtcagatcgc ctggagacgc
     2821 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccggactcta
     2881 gcgtttaaac ttaagcttgg taccgagctc ggatccacta gtccagtgtg gtggaattct
     2941 gcagatatca acaagtttgt acaaaaaagt tggcatgagc gccgtgaaga gaaccatcaa
     3001 gggaacccac cactggctgc tgctgaccat cctgacctcc ctgctggtgc tggtgcagtc
     3061 tacccagtgg agcctgttct tcttcctgta cgagaacgcc ttcctgccct tcgctatggg
     3121 tatcatcgcc atgtccgcct tcgctatgat gttcgtgaag cacaagcacg ctttcctgtg
     3181 cctgttcctg ctgccatccc tggccaccgt ggcttacttc aacatggtgt acatgcccgc
     3241 ctcttgggtc atgcgcatca tgacctggct ggacatggtg gacaccagcc tgaagctgaa
     3301 ggactgcgtg atgtacgcct ctgctgtggt gctgctgatc ctgatgaccg ctagaaccgt
     3361 gtacgatgat ggtgctagaa gggtgtggac cctgatgaac gtgctgaccc tggtgtacaa
     3421 ggtgtactac ggcaacgccc tggaccaggc tatcagcatg tgggctctga tcatctctgt
     3481 gaccagcaac tactccggag tggtgaccac cgtgatgttc ctggccagag gtatcgtgtt
     3541 catgtgcgtg gagtactgcc ctatcttctt catcaccggc aacaccctgc agtgcatcat
     3601 gctggtgtac tgcttcctgg gttacttctg cacctgctat ttcggcctgt tctgcctgct
     3661 gaacagatac ttccgtctga ccctgggcgt gtacgactac ctggtgtcca cccaggaatt
     3721 ccgttacatg aactctcagg gactgctgcc ccctaagaac agcatcgacg ctttcaagct
     3781 gaacatcaag ctgctgggag tgggcggaaa gccatgcatc aaggtggcca ccgtgcagta
     3841 cccaactttc ttgtacaaag tggttgatat caaggccgcc gacatcacca gcctgtacaa
     3901 gaaagtgggg tggagccatc cccagttcga gaagggcgga ggatctggcg gcggaagcgg
     3961 cggaggaagc tggtcccacc ctcagtttga gaaaggcacc gagctgggca gcaccatggc
     4021 cagctacccc tacgacgtgc ccgactacgc caaggacaac accgtgcccc tgaagctgat
     4081 cgccctgctg gccaacggcg agttccactc tggcgagcag ctgggcgaga cactgggcat
     4141 gagcagagcc gccatcaaca agcacatcca gaccctgcgg gactggggcg tggacgtgtt
     4201 cacagtgccc ggcaagggct acagcctgcc cgagcccatc cagctgctga acgccaagca
     4261 gatcctgggc cagctggatg gcggatctgt ggccgtgctg cccgtgatcg acagcaccaa
     4321 ccagtatctg ctcgaccgga tcggcgagct gaagtccggc gacgcctgta tcgccgagta
     4381 ccagcaggcc ggcagaggcg gcaggggcag aaagtggttc agccccttcg gcgccaacct
     4441 gtacctgagc atgttctgga gactggaaca gggcccagcc gccgctattg gcctgagcct
     4501 ggtcatcggc atcgtgatgg ccgaggtgct gcggaagctg ggcgccgaca aagtgcgcgt
     4561 gaagtggccc aacgacctgt atctgcagga ccggaagctg gccggcatcc tggtggaact
     4621 gaccggcaaa acaggcgacg ccgcccagat cgtgatcggc gctggcatca acatggccat
     4681 gcggcgggtg gaagagagcg tggtcaacca gggctggatc accctgcagg aagccggcat
     4741 caacctggac cggaacaccc tggccgccat gctgatcaga gagctgagag ccgccctgga
     4801 actgttcgag caggaaggcc tggcccccta cctgagcaga tgggagaagc tggacaactt
     4861 catcaaccgg cccgtgaaac tgatcatcgg cgacaaagag atcttcggca tcagccgggg
     4921 catcgacaag cagggggccc tgctcctgga acaggacggc atcatcaagc cctggatggg
     4981 cggcgagatc agcctgcgga gcgccgagaa gtgatgactc gagtctagag ggcccgttta
     5041 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     5101 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     5161 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     5221 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     5281 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     5341 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5401 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5461 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5521 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtacctaga agttcctatt
     5581 ccgaagttcc tattctctag aaagtatagg aacttccttg gccaaaaagc ctgaactcac
     5641 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     5701 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     5761 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     5821 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct
     5881 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     5941 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
     6001 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     6061 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     6121 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     6181 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     6241 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     6301 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     6361 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct
     6421 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     6481 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     6541 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     6601 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tactacgaga
     6661 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6721 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     6781 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6841 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6901 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     6961 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7021 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7081 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7141 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactga
//
