LOCUS       sequence-386170-        7026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sars-CoV-2 ORF3a_Q57H in MAC-C destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7026)
  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 7026)
  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: MAC-C Sars-CoV-2 ORF3a_Q57H
FEATURES             Location/Qualifiers
     source          1..7026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(110..127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(281..869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(361..380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1040..1831,1832..1900))
                     /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     1663..1682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1901..2005)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2080..2099)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2271..2650
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2651..2854
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2805..2825
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2856..2874
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2877..2895
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2905..2929
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3072..3096
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3928..3952)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(4011..4358,4359..4361,4362..4970)
                     /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(5014..5031)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5020..5244
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5377..5396)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5528..5575
                     /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             5583..6605
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             5583..6605
                     /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    6735..6856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6772..6791)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6826..6845
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6905..6921)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6905..6921)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6918..6940)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6929..6945
                     /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(6953..6959,6960..6977,6978..6983))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6998..7019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
       61 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
      121 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
      181 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
      241 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
      301 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
      361 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
      421 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
      481 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
      541 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
      601 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
      661 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
      721 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
      781 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
      841 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
      901 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
      961 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1021 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1081 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1141 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     1201 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1261 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1321 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1381 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1441 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1501 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1561 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1621 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1681 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1741 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     1801 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1861 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     1921 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     1981 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     2041 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     2101 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     2161 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     2221 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
     2281 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     2341 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2401 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2461 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2521 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2581 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2641 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2701 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2761 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2821 gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata gagatctccc
     2881 tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg cctggagacg
     2941 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
     3001 agcgtttaaa cttaagcttg gtaccgagct cggatccact agtccagtgt ggtggaattc
     3061 tgcagatatc aacaagtttg tacaaaaaag caggctccac catggacctt tttatgagaa
     3121 tctttaccat cgggactgtc acactgaaac agggagaaat taaggacgcg accccgagtg
     3181 actttgtgag ggccactgct acaattccca tccaagcctc tcttccattc ggctggctca
     3241 tagtgggtgt ggcactgctc gcggtattcc actcagcgtc taagatcata acgctgaaga
     3301 agagatggca gttggctctc agcaaaggtg ttcatttcgt ctgtaacctg cttcttcttt
     3361 tcgtcactgt ctattctcac ttgctcttgg ttgcagccgg gctggaagcc ccgtttctgt
     3421 atctgtacgc gctcgtttac ttcctccaga gcataaattt cgtacgaatt attatgagac
     3481 tctggttgtg ctggaagtgt cggagcaaga atcctctctt gtatgacgcc aattacttcc
     3541 tttgttggca cactaattgt tatgattact gcatacctta caatagcgtt acctcatcca
     3601 tagtaatcac ctccggtgac ggaacaacgt caccaatctc cgagcatgat taccagattg
     3661 gaggctatac ggaaaaatgg gagagtggtg tcaaggactg tgtggtgctg catagctatt
     3721 tcacaagcga ttattatcaa ctgtacagta cacagttgtc cacggatacc ggggtcgagc
     3781 atgtcacatt ttttatttat aacaagatcg ttgatgagcc agaggagcat gtgcagatcc
     3841 acacgataga tggaagctcc ggtgtagtga atccagtcat ggaaccgatc tatgacgagc
     3901 ccaccacgac cacatctgtg cccctggacc cagctttctt gtacaaagtg grmgktatca
     3961 agmwggrcwg caycayrryc wgytacmmck amgasgtgmc cgwctacgcc aaggacaaca
     4021 ccgtgcccct gaagctgatc gccctgctgg ccaacggcga gttccactct ggcgagcagc
     4081 tgggcgagac actgggcatg agcagagccg ccatcaacaa gcacatccag accctgcggg
     4141 actggggcgt ggacgtgttc acagtgcccg gcaagggcta cagcctgccc gagcccatcc
     4201 agctgctgaa cgccaagcag atcctgggcc agctggatgg cggatctgtg gccgtgctgc
     4261 ccgtgatcga cagcaccaac cagtatctgc tcgaccggat cggcgagctg aagtccggcg
     4321 acgcctgtat cgccgagtac cagcaggccg gcagaggcgg caggggcaga aagtggttca
     4381 gccccttcgg cgccaacctg tacctgagca tgttctggag actggaacag ggcccagccg
     4441 ccgctattgg cctgagcctg gtcatcggca tcgtgatggc cgaggtgctg cggaagctgg
     4501 gcgccgacaa agtgcgcgtg aagtggccca acgacctgta tctgcaggac cggaagctgg
     4561 ccggcatcct ggtggaactg accggcaaaa caggcgacgc cgcccagatc gtgatcggcg
     4621 ctggcatcaa catggccatg cggcgggtgg aagagagcgt ggtcaaccag ggctggatca
     4681 ccctgcagga agccggcatc aacctggacc ggaacaccct ggccgccatg ctgatcagag
     4741 agctgagagc cgccctggaa ctgttcgagc aggaaggcct ggccccctac ctgagcagat
     4801 gggagaagct ggacaacttc atcaaccggc ccgtgaaact gatcatcggc gacaaagaga
     4861 tcttcggcat cagccggggc atcgacaagc agggggccct gctcctggaa caggacggca
     4921 tcatcaagcc ctggatgggc ggcgagatca gcctgcggag cgccgagaag tgatgactcg
     4981 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     5041 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     5101 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     5161 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     5221 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     5281 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5341 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5401 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5461 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5521 gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga acttccttgg
     5581 ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     5641 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     5701 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     5761 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
     5821 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     5881 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg
     5941 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     6001 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
     6061 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
     6121 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     6181 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     6241 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     6301 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
     6361 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
     6421 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
     6481 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
     6541 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
     6601 aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6661 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6721 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6781 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6841 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     6901 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6961 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     7021 ttgcgt
//
