LOCUS       sequence-346793-        8598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  501V2 variant S-protein with c-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8598)
  TITLE     Spike variants with FLAG tags
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8598)
  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            "Unpublished"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: pCAGGS-S-501V2-FLAG
FEATURES             Location/Qualifiers
     source          1..8598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          663..1683
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1691..1710
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             complement(3056..3073)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             5569..5592
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5620..5639)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5685..5740
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5739..5758)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6101..6117)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6101..6117)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6114..6136)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6125..6141
                     /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(6149..6155,6156..6173,6174..6179))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6194..6215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6274..6469
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6320..6455
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6475..6609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6512..6531)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6566..6585
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6677..6694)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6848..7436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6928..6947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7607..8398,8399..8467))
                     /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     8230..8249
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8468..8572)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggnsgcg cgccrggggg gggggggsgg ggsgrggggs
      541 ggggsggggs grggsggaga ggkgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg grgggctcgg gggrggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagctag agcctctgct aaccatgttc atgccttctt ctttttccta
     1681 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcgga
     1741 tccatgtttg tgttcctggt gctgctgcca ctggtgtcca gccagtgtgt gaacttcacc
     1801 accaggaccc aacttcctcc tgcctacacc aactccttca ccaggggagt ctactaccct
     1861 gacaaggtgt tcaggtcctc tgtgctgcac agcacccagg acctgttcct gccattcttc
     1921 agcaatgtga cctggttcca tgccatccat gtgtctggca ccaatggcac caagaggttt
     1981 gccaaccctg tgctgccatt caatgatgga gtctactttg ccagcacaga gaagagcaac
     2041 atcatcaggg gctggatttt tggcaccacc ctggacagca agacccagtc cctgctgatt
     2101 gtgaacaatg ccaccaatgt ggtgattaag gtgtgtgagt tccagttctg taatgaccca
     2161 ttcctgggag tctactacca caagaacaac aagtcctgga tggagtctga gttcagggtc
     2221 tactcctctg ccaacaactg tacctttgaa tatgtgagcc aaccattcct gatggacttg
     2281 gagggcaagc agggcaactt caagaacctg agggagtttg tgttcaagaa cattgatggc
     2341 tacttcaaga tttacagcaa acacacacca atcaacctgg tgaggggcct gccacagggc
     2401 ttctctgcct tggaaccact ggtggacctg ccaattggca tcaacatcac caggttccag
     2461 accctgctgg ctctgcacat ctcctacctg acacctggag actcctcctc tggctggaca
     2521 gcaggagcag cagcctacta tgtgggctac ctccaaccaa ggaccttcct gctgaaatac
     2581 aatgagaatg gcaccatcac agatgctgtg gactgtgccc tggacccact gtctgagacc
     2641 aagtgtaccc tgaaatcctt cacagtggag aagggcatct accagaccag caacttcagg
     2701 gtccaaccaa cagagagcat tgtgaggttt ccaaacatca ccaacctgtg tccatttgga
     2761 gaggtgttca atgccaccag gtttgcctct gtctatgcct ggaacaggaa gaggattagc
     2821 aactgtgtgg ctgactactc tgtgctctac aactctgcct ccttcagcac cttcaagtgt
     2881 tatggagtga gcccaaccaa actgaatgac ctgtgtttca ccaatgtcta tgctgactcc
     2941 tttgtgatta ggggagatga ggtgagacag attgcccctg gacaaacagg caacattgct
     3001 gactacaact acaaactgcc tgatgacttc acaggctgtg tgattgcctg gaacagcaac
     3061 aacctggaca gcaaggtggg aggcaactac aactacctct acagactgtt caggaagagc
     3121 aacctgaaac catttgagag ggacatcagc acagagattt accaggctgg cagcacacca
     3181 tgtaatggag tgaagggctt caactgttac tttccactcc aatcctatgg cttccaacca
     3241 acctatggag tgggctacca accatacagg gtggtggtgc tgtcctttga actgctccat
     3301 gcccctgcca cagtgtgtgg accaaagaag agcaccaacc tggtgaagaa caagtgtgtg
     3361 aacttcaact tcaatggact gacaggcaca ggagtgctga cagagagcaa caagaagttc
     3421 ctgccattcc aacagtttgg cagggacatt gctgacacca cagatgctgt gagggaccca
     3481 cagaccttgg agattctgga catcacacca tgttcctttg gaggagtgtc tgtgattaca
     3541 cctggcacca acaccagcaa ccaggtggct gtgctctacc agggtgtgaa ctgtactgag
     3601 gtgcctgtgg ctatccatgc tgaccaactt acaccaacct ggagggtcta cagcacaggc
     3661 agcaatgtgt tccagaccag ggctggctgt ctgattggag cagagcatgt gaacaactcc
     3721 tatgagtgtg acatcccaat tggagcaggc atctgtgcct cctaccagac ccagaccaac
     3781 agcccaagga gggcaaggtc tgtggcaagc cagagcatca ttgcctacac aatgagtctg
     3841 ggagtagaga actctgtggc ttacagcaac aacagcattg ccatcccaac caacttcacc
     3901 atctctgtga ccacagagat tctgcctgtg agtatgacca agacctctgt ggactgtaca
     3961 atgtatatct gtggagacag cacagagtgt agcaacctgc tgctccaata tggctccttc
     4021 tgtacccaac ttaacagggc tctgacaggc attgctgtgg aacaggacaa gaacacccag
     4081 gaggtgtttg cccaggtgaa gcagatttac aagacacctc caatcaagga ctttggaggc
     4141 ttcaacttca gccagattct gcctgaccca agcaagccaa gcaagaggtc cttcattgag
     4201 gacctgctgt tcaacaaggt gaccctggct gatgctggct tcatcaagca atatggagac
     4261 tgtctgggag acattgctgc cagggacctg atttgtgccc agaagttcaa tggactgaca
     4321 gtgctgcctc cactgctgac agatgagatg attgcccaat acacctctgc cctgctggct
     4381 ggcaccatca cctctggctg gacctttgga gcaggagcag ccctccaaat cccatttgct
     4441 atgcagatgg cttacaggtt caatggcatt ggagtgaccc agaatgtgct ctatgagaac
     4501 cagaaactga ttgccaacca gttcaactct gccattggca agattcagga ctccctgtcc
     4561 agcacagcct ctgccctggg caaactccaa gatgtggtga accagaatgc ccaggctctg
     4621 aacaccctgg tgaagcaact ttccagcaac tttggagcca tctcctctgt gctgaatgac
     4681 atcctgagca gactggacaa ggtggaggct gaggtccaga ttgacagact gattacaggc
     4741 agactccaat ccctccaaac ctatgtgacc caacaactta tcagggctgc tgagattagg
     4801 gcatctgcca acctggctgc caccaagatg agtgagtgtg tgctgggaca aagcaagagg
     4861 gtggacttct gtggcaaggg ctaccacctg atgagttttc cacagtctgc ccctcatgga
     4921 gtggtgttcc tgcatgtgac ctatgtgcct gcccaggaga agaacttcac cacagcccct
     4981 gccatctgcc atgatggcaa ggctcacttt ccaagggagg gagtgtttgt gagcaatggc
     5041 acccactggt ttgtgaccca gaggaacttc tatgaaccac agattatcac cacagacaac
     5101 acctttgtgt ctggcaactg tgatgtggtg attggcattg tgaacaacac agtctatgac
     5161 ccactccaac ctgaactgga ctccttcaag gaggaactgg acaaatactt caagaaccac
     5221 accagccctg atgtggacct gggagacatc tctggcatca atgcctctgt ggtgaacatc
     5281 cagaaggaga ttgacagact gaatgaggtg gctaagaacc tgaatgagtc cctgattgac
     5341 ctccaagaac tgggcaaata tgaacaatac atcaagtggc catggtacat ctggctgggc
     5401 ttcattgctg gactgattgc cattgtgatg gtsaccatca tgctgtgctg catgacctca
     5461 tgttgctcct gtcttaaggg gtgctgctcc tgcggttcct gttgcaaatt tgacgaggac
     5521 gacagcgaac cggtgcttaa gggtgtgaag ctgcattaca ccggaggcga ttacaaggat
     5581 gacgatgaca agtgacggcc gcctgcaggt taattaagat ctttttccct ctgccaaaaa
     5641 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     5701 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     5761 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     5821 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
     5881 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     5941 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     6001 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     6061 gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6121 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6181 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6241 tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt
     6301 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6361 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6421 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg
     6481 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6541 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6601 gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6661 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6721 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6781 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6841 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6901 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6961 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7021 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7081 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7141 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7201 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7261 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7501 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7561 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7621 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7681 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7741 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7801 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7861 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7921 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7981 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8041 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8101 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8161 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8221 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8281 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8341 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8401 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8461 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8521 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8581 cccgaaaagt gccacctg
//
