LOCUS       sequence-384456-        5664 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sars-CoV-2 E_P71L in MAC-C destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5664)
  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 5664)
  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: "10 May 
            2024, Research Square"
COMMENT     Sequence Label: HA-C Sars-CoV-2 E P71L
FEATURES             Location/Qualifiers
     source          1..5664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1077..1198
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1114..1133)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1168..1187
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1247..1263)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1247..1263)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1260..1282)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1271..1287
                     /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(1295..1301,1302..1319,1320..1325))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1340..1361
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1478..1495)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1649..2237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1729..1748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2408..3199,3200..3268))
                     /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     3031..3050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3269..3373)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3448..3467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3639..4018
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4019..4222
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4173..4193
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4224..4242
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4245..4263
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4273..4297
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    4440..4464
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4765..4789)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4836..4859
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4896..4919
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4950..4976
                     /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             4950..4976
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5020..5037)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5026..5250
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5383..5402)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5534..5581
                     /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             join(5589..5664,1..947)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             join(5589..5664,1..947)
                     /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
                     "
ORIGIN
        1 agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg
       61 tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact
      121 ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc
      181 tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg
      241 aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc
      301 ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat
      361 ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg
      421 acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg
      481 actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg
      541 acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat
      601 acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc
      661 gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc
      721 tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag
      781 cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta
      841 cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg
      901 atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac gtactacgag
      961 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     1021 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     1081 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     1141 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     1201 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     1261 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1321 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1381 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1441 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1501 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1561 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1621 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1681 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1741 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1801 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1861 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1921 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1981 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2041 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2101 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2161 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2221 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2281 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2341 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2401 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2461 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2521 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2581 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2641 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2701 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2761 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2821 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2881 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2941 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3001 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3061 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3121 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3181 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3241 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3301 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3361 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     3421 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3481 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     3541 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     3601 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
     3661 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     3721 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     3781 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     3841 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     3901 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     3961 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     4021 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     4081 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     4141 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     4201 ggaggtctat ataagcagag ctctccctat cagtgataga gatctcccta tcagtgatag
     4261 agatcgtcga cgagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
     4321 ttttgacctc catagaagac accgggaccg atccagcctc cggactctag cgtttaaact
     4381 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcaa
     4441 caagtttgta caaaaaagca ggctccacca tgtacagctt cgtatcagaa gaaaccggga
     4501 cactgatcgt aaattctgtg ctcttgtttc tggcattcgt cgtatttctc ctcgtcacac
     4561 tggcaattct gactgcattg aggctttgcg cctactgttg taacattgtc aatgtatctc
     4621 tcgtgaaacc ctcattctac gtttacagca gggtgaagaa tctcaattct agcagggtgc
     4681 tggatctcct cgttgacccc tcattctacg tttacagcag ggtgaagaat ctcaattcta
     4741 gcagggtgct ggatctcctc gttgacccag ctttcttgta caaagtggtt gatatcaagg
     4801 ccgccgacat caccagcctg tacaagaaag tggggtggag ccatccccag ttcgagaagg
     4861 gcggaggatc tggcggcgga agcggcggag gaagctggtc ccaccctcag tttgagaaag
     4921 gcaccgagct gggcagcacc atggccagct acccctacga cgtgcccgac tacgcctgat
     4981 gactcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     5041 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     5101 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     5161 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     5221 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     5281 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5341 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5401 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5461 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5521 gttcacgtac ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt
     5581 ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     5641 tcgacagcgt ctccgacctg atgc
//
