LOCUS       sequence-437731-        9194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pseudovirus plasmid that expresses spike of EG.5.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9194)
  TITLE     SARS-CoV-2 variant
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9194)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3.3_SARS2_EG.5.1
FEATURES             Location/Qualifiers
     source          1..9194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1584..1608)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1605..1808)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1634..1654)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1809..2188
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2360..2379
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2454..2558
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2559..2627,2628..3419)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRINAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2777..2796)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3590..4178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4079..4098
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4332..4349
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4466..4487
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4502..4507,4508..4525,4526..4532)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4540..4556
                     /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)."
     primer_bind     4545..4567
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4564..4580
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4564..4580
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4629..4750
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4640..4659)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4694..4713
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4926..5720)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5037..5056)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5647..5666
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5787..6116)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5801..5936
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5855..5874)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6101..6121
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(6130..6558)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6240..6261)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6452..6471
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6760..6808
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     6833..6852
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
ORIGIN
        1 aagccatagg actggagtgg agagtaacag ttcgggcccg ccactccatt acatggtttg
       61 ttgccagcct ggtaaatctc tgtgctgatg tccctctcaa atggtttcag cttgctcttc
      121 ctgagcagtc tgtagaggta gttgtagttg cctgacggct tgctgtccag cttgttgctg
      181 ttccaggcaa tcacacagcc tgtgaagtca tcaggcagtt tgtagttgta gtcagcaatg
      241 ttgcctgttt gtccaggggc aatctgactc acctcatttc ccctaatcac aaaggagtca
      301 gcatagacat tggtgaaaca caggtcattc agtttggttg ggctcactcc ataacacttg
      361 aaggcgaaga agggggcaaa gttgtagatc acagagtagt cagccacaca gttgctaatc
      421 ctcttcctgt tccaggcata gacagaggca aacgtggtgg cattgaacac ctcgtgaaat
      481 ggacacaggt tggtgatgtt tggaaacctc acaatgctct ctgttggttg gaccctgaag
      541 ttgctggtct ggtagatgcc cttctccact gtgaaggatt tcagggtaca cttggtctca
      601 gacagtgggt ccagggcaca gtccacagca tctgtgatgg tgccattctc attgtatttc
      661 agcaggaagg tccttggttg gaggtagccc acatagtagg ctgctgctcc tgctgtccag
      721 ccagaggagg agtctacagg tgtcaggtag gacctgtgca gagccagcag ggtctggaac
      781 ctggtgatgt tgatgccaat tggcaggtcc accagtggtt ccaaggcaga gaagccctgt
      841 ggcaggtccc tctccaggtt gattggtgtg tgtttgctgt aaatcttgaa gtagccatca
      901 atgttcttga acacaaactc cctcaggttc ttgaagttgc cctccttgcc ctccaagtcc
      961 atcaggaatg gttggctcac atattcaaag gtacagttgt tggcagagga gtagaccctg
     1021 aactcagact ccatccagga cttgttgttc ttctggtaga cgtccaggaa tgggtcatta
     1081 cagaactgga actcacacac cttaatcacc acattggtgg cattgttcac aatcagcagg
     1141 gactgggtct tgctgtccag ggtggtgcca aaaatccagc ccctgatgat gttgctcttc
     1201 tctgtgctgg caaagtagac tccatcattg aatggcagcg cagggttgtc aaacctcttg
     1261 gtgccattgg tgccagacac atggatggca tggaaccagg tcacattgct gaagaatggc
     1321 aggaacaggt cgtgggtgct gtgcagcaca gaggacctga acaccttgtc agggtagtag
     1381 actcccctgg tgaaggagtt ggtgtaggat tgggtcctgg tgatcaggtt cacacactgg
     1441 ctggacacca gtggcagcag caccaggaac acaaacatgg atccgaattc tctagagtta
     1501 accttaagga gctcagtccg gaggctggat cggtcccggt gtcttctatg gaggtcaaaa
     1561 cagcgtggat ggcgtctcca ggcgatctga cggttcacta aacgagctct gcttatatag
     1621 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
     1681 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
     1741 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
     1801 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     1861 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     1921 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     1981 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     2041 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     2101 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     2161 tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac atcgcgaagc
     2221 agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt
     2281 gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca agcagggagc
     2341 agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagggga
     2401 tcgggagatc tcccgatccg tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2461 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2521 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2581 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2641 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2701 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2761 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattaacgcc gggcaagagc
     2821 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2881 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2941 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3001 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3061 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3121 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3181 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3241 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3301 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3361 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3421 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3481 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3541 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3601 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3661 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3721 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3781 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3841 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3901 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3961 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4021 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4081 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4141 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4201 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4261 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4321 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     4381 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4441 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     4501 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     4561 acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc gacggtatac
     4621 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4681 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4741 taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     4801 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     4861 cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgcga accccagagt
     4921 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     4981 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     5041 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     5101 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     5161 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
     5221 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     5281 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     5341 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     5401 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     5461 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     5521 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     5581 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     5641 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     5701 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatccgaaa
     5761 atggatatac aagctcccgg gagctttttg caaaagccta ggcctccaaa aaagcctcct
     5821 cactacttct ggaatagctc agaggcagag gcggcctcgg cctctgcata aataaaaaaa
     5881 attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
     5941 ggagttaggg gcgggactat ggttgctgac taattgagat gtatgctttg catacttctg
     6001 cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt
     6061 tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc
     6121 cacagaatta attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     6181 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     6241 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     6301 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     6361 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     6421 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     6481 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     6541 cgccgctaca gggcgcgtgg ggataccccc tagagcccca gctgcgcaga tctgctatgg
     6601 cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt
     6661 ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat
     6721 cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacacccg
     6781 tgcgttttat tctgtctttt tattgccgtc atagcgcggg ttccttccgg tattgtctcc
     6841 ttccgtgttt cagttagcct cccccgtttg gtaccaccgg ttcagtcagt tagatatcaa
     6901 gcttccatgg ctcgagtcac ttacaacagg agccacagga acaacagcct ttcagacagg
     6961 aacaacagga ggtcatacaa cacagcatta tggtcaccat cacaatggca atcagtccag
     7021 caatgaagcc cagccagatg taccatggcc acttgatgta ttgttcatat ttgcccagtt
     7081 cttggaggtc aatcagggac tcattcaggt tcttagccac ctcattcagt ctgtcaatct
     7141 ccttctggat gttcaccaca gaggcattga tgccagagat gtctcccagg tccacatcag
     7201 ggctggtgtg gttcttgaag tatttgtcca gttcctcctt gaaggagtcc agttcaggtt
     7261 ggagtgggtc atagactgtg ttgttcacaa tgccaatcac cacatcacag ttgccagaca
     7321 caaaggtgtt gtctgtggtg ataatctgtg gttcatagaa gttcctctgg gtcacaaacc
     7381 agtgggtgcc attgctcaca aacactccct cccttggaaa gtgagccttg ccatcatggc
     7441 agatggcagg ggctgtggtg aagttcttct cctgggcagg cacataggtc acatgcagga
     7501 acaccactcc atgaggggca gactgtggaa aactcatcag gtggtagccc ttgccacaga
     7561 agtccaccct cttgctttgt cccagcacac actcactcat cttggtggca gccaggttgg
     7621 cagatgccct aatctcagca gccctgataa gttgttgggt cacataggtt tggagggatt
     7681 ggagtctgcc tgtaatcagt ctgtcaatct ggacctcagc ctccaccttg tccagtctgc
     7741 tcaggatgtc attcagcaca gaggagatgg ctccaaactt gctggaaagt tgcttcacca
     7801 gggtgttcag agcctgggca ttatggttca ccacatcttg gagtttgccc agggcagagg
     7861 ctgtgctgga cagggagtcc tgaatcttgc caatggcaga gttgaactgg ttggcaatca
     7921 gtttctggtt ctcatagagc acattctggg tcactccaat gccattgaac ctgtaagcca
     7981 tctgcatagc aaatgggatt tggagggctg ctcctgctcc aaaggtccag ccagaggtga
     8041 tggtgccagc cagcagggca gaggtgtatt gggcaatcat ctcatctgtc agcagtggag
     8101 gcagcacggt cagtccattg aacttctggg cacaaatcag gtcccttgca gcaatgtctc
     8161 ccagacagtc tccatattgc ttgatgaagc cagcatcagc cagggtcacc ttgttgaaca
     8221 gcaggtcctc aatgaaggac ctcttgcttg gcttgcttgg gtcaggcaga atctggctga
     8281 agttgaagcc tccaaagtac ttgattggag gtgtcttgta aatctgcttc acctgggcaa
     8341 acacctcctg ggtgttcttg tcctgttcca cagcaatgcc tgtcagagcc ctcttaagtt
     8401 gggtacagaa ggagccatat tggagcagca ggttgctaca ctctgtgctg tctccacaga
     8461 tatacattgt acagtccaca gaggtcttgg tcatactcac aggcagaatt tctgtggtca
     8521 cagagatggt gaagttggtt gggatggcaa tgctgttgtt gctgtaagcc acagagttct
     8581 ctgctcccag actcattgtg taggcaatga tgctctggct tgccacagac cttgccctcc
     8641 tatggctctt ggtctgggtc tggtaggagg cacagatgcc tgctccaatt gggatgtcac
     8701 actcatagga gttgttcaca tactctgctc caatcagaca gccagccctg gtctggaaca
     8761 cattgctgcc tgtgctgtag accctccagg ttggtgtaag ttggtcagca tggatagcca
     8821 caggcacctc agtacagttc acaccctggt agagcacagc cacctggttg ctggtgttgg
     8881 tgccaggtgt aatcacagac actcctccaa aggaacatgg tgtgatgtcc agaatctcca
     8941 aggtctgtgg gtccctcaca gcatctgtgg tgtcagcaat gtccctgcca aactgttgga
     9001 atggcaggaa cttcttgttg ctctctgtca gcactcctgt gcctgtcagt ccattgaagt
     9061 tgaagttcac acacttgttc ttcaccaggt tggtgctctt ctttggtcca cacactgtgg
     9121 caggggcatg gagcagttca aaggacagca ccaccaccct gtatggttgg tggcccactc
     9181 cataggttgg tctg
//
