LOCUS       sequence-346798-        8589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  B.1.1.7 variant S-protein with c-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8589)
  TITLE     Spike variants with FLAG tags
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8589)
  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            "Unpublished"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-B.1.1.7-FLAG
FEATURES             Location/Qualifiers
     source          1..8589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1682
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1690..1709
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             complement(3046..3063)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             5559..5582
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5610..5629)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5675..5730
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5729..5748)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6091..6107)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6091..6107)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6104..6126)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6115..6131
                     /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(6139..6145,6146..6163,6164..6169))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6184..6205
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6264..6459
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6310..6445
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6465..6599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6502..6521)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6556..6575
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6667..6684)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6838..7426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6918..6937)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7597..8388,8389..8457))
                     /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     8220..8239
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8458..8562)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggnsgcgc gccrggsggg gsggggsggg gsgrggggsg
      541 gggsggggsg rggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg rgggctcggg ggrggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     1681 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcggat
     1741 ccatgtttgt gttcctggtg ctgctgccac tggtgtccag ccagtgtgtg aacctgacca
     1801 ccaggaccca acttcctcct gcctacacca actccttcac caggggagtc tactaccctg
     1861 acaaggtgtt caggtcctct gtgctgcaca gcacccagga cctgttcctg ccattcttca
     1921 gcaatgtgac ctggttccat gccatctctg gcaccaatgg caccaagagg tttgacaacc
     1981 ctgtgctgcc attcaatgat ggagtctact ttgccagcac agagaagagc aacatcatca
     2041 ggggctggat ttttggcacc accctggaca gcaagaccca gtccctgctg attgtgaaca
     2101 atgccaccaa tgtggtgatt aaggtgtgtg agttccagtt ctgtaatgac ccattcctgg
     2161 gagtctacca caagaacaac aagtcctgga tggagtctga gttcagggtc tactcctctg
     2221 ccaacaactg tacctttgaa tatgtgagcc aaccattcct gatggacttg gagggcaagc
     2281 agggcaactt caagaacctg agggagtttg tgttcaagaa cattgatggc tacttcaaga
     2341 tttacagcaa acacacacca atcaacctgg tgagggacct gccacagggc ttctctgcct
     2401 tggaaccact ggtggacctg ccaattggca tcaacatcac caggttccag accctgctgg
     2461 ctctgcacag gtcctacctg acacctggag actcctcctc tggctggaca gcaggagcag
     2521 cagcctacta tgtgggctac ctccaaccaa ggaccttcct gctgaaatac aatgagaatg
     2581 gcaccatcac agatgctgtg gactgtgccc tggacccact gtctgagacc aagtgtaccc
     2641 tgaaatcctt cacagtggag aagggcatct accagaccag caacttcagg gtccaaccaa
     2701 cagagagcat tgtgaggttt ccaaacatca ccaacctgtg tccatttgga gaggtgttca
     2761 atgccaccag gtttgcctct gtctatgcct ggaacaggaa gaggattagc aactgtgtgg
     2821 ctgactactc tgtgctctac aactctgcct ccttcagcac cttcaagtgt tatggagtga
     2881 gcccaaccaa actgaatgac ctgtgtttca ccaatgtcta tgctgactcc tttgtgatta
     2941 ggggagatga ggtgagacag attgcccctg gacaaacagg caagattgct gactacaact
     3001 acaaactgcc tgatgacttc acaggctgtg tgattgcctg gaacagcaac aacctggaca
     3061 gcaaggtggg aggcaactac aactacctct acagactgtt caggaagagc aacctgaaac
     3121 catttgagag ggacatcagc acagagattt accaggctgg cagcacacca tgtaatggag
     3181 tggagggctt caactgttac tttccactcc aatcctatgg cttccaacca acctatggag
     3241 tgggctacca accatacagg gtggtggtgc tgtcctttga actgctccat gcccctgcca
     3301 cagtgtgtgg accaaagaag agcaccaacc tggtgaagaa caagtgtgtg aacttcaact
     3361 tcaatggact gacaggcaca ggagtgctga cagagagcaa caagaagttc ctgccattcc
     3421 aacagtttgg cagggacatt gatgacacca cagatgctgt gagggaccca cagaccttgg
     3481 agattctgga catcacacca tgttcctttg gaggagtgtc tgtgattaca cctggcacca
     3541 acaccagcaa ccaggtggct gtgctctacc agggtgtgaa ctgtactgag gtgcctgtgg
     3601 ctatccatgc tgaccaactt acaccaacct ggagggtcta cagcacaggc agcaatgtgt
     3661 tccagaccag ggctggctgt ctgattggag cagagcatgt gaacaactcc tatgagtgtg
     3721 acatcccaat tggagcaggc atctgtgcct cctaccagac ccagaccaac agccatagga
     3781 gggcaaggtc tgtggcaagc cagagcatca ttgcctacac aatgagtctg ggagcagaga
     3841 actctgtggc ttacagcaac aacagcattg ccatcccaat caacttcacc atctctgtga
     3901 ccacagagat tctgcctgtg agtatgacca agacctctgt ggactgtaca atgtatatct
     3961 gtggagacag cacagagtgt agcaacctgc tgctccaata tggctccttc tgtacccaac
     4021 ttaacagggc tctgacaggc attgctgtgg aacaggacaa gaacacccag gaggtgtttg
     4081 cccaggtgaa gcagatttac aagacacctc caatcaagga ctttggaggc ttcaacttca
     4141 gccagattct gcctgaccca agcaagccaa gcaagaggtc cttcattgag gacctgctgt
     4201 tcaacaaggt gaccctggct gatgctggct tcatcaagca atatggagac tgtctgggag
     4261 acattgctgc cagggacctg atttgtgccc agaagttcaa tggactgaca gtgctgcctc
     4321 cactgctgac agatgagatg attgcccaat acacctctgc cctgctggct ggcaccatca
     4381 cctctggctg gacctttgga gcaggagcag ccctccaaat cccatttgct atgcagatgg
     4441 cttacaggtt caatggcatt ggagtgaccc agaatgtgct ctatgagaac cagaaactga
     4501 ttgccaacca gttcaactct gccattggca agattcagga ctccctgtcc agcacagcct
     4561 ctgccctggg caaactccaa gatgtggtga accagaatgc ccaggctctg aacaccctgg
     4621 tgaagcaact ttccagcaac tttggagcca tctcctctgt gctgaatgac atcctggcca
     4681 gactggacaa ggtggaggct gaggtccaga ttgacagact gattacaggc agactccaat
     4741 ccctccaaac ctatgtgacc caacaactta tcagggctgc tgagattagg gcatctgcca
     4801 acctggctgc caccaagatg agtgagtgtg tgctgggaca aagcaagagg gtggacttct
     4861 gtggcaaggg ctaccacctg atgagttttc cacagtctgc ccctcatgga gtggtgttcc
     4921 tgcatgtgac ctatgtgcct gcccaggaga agaacttcac cacagcccct gccatctgcc
     4981 atgatggcaa ggctcacttt ccaagggagg gagtgtttgt gagcaatggc acccactggt
     5041 ttgtgaccca gaggaacttc tatgaaccac agattatcac cacacacaac acctttgtgt
     5101 ctggcaactg tgatgtggtg attggcattg tgaacaacac agtctatgac ccactccaac
     5161 ctgaactgga ctccttcaag gaggaactgg acaaatactt caagaaccac accagccctg
     5221 atgtggacct gggagacatc tctggcatca atgcctctgt ggtgaacatc cagaaggaga
     5281 ttgacagact gaatgaggtg gctaagaacc tgaatgagtc cctgattgac ctccaagaac
     5341 tgggcaaata tgaacaatac atcaagtggc catggtacat ctggctgggc ttcattgctg
     5401 gactgattgc cattgtgatg gtgaccatca tgctgtgctg catgacctca tgttgctcct
     5461 gtcttaaggg gtgctgctcc tgcggttcct gttgcaaatt tgacgaggac gacagcgaac
     5521 cggtgcttaa gggtgtgaag ctgcattaca ccggaggcga ttacaaggat gacgatgaca
     5581 agtgacggcc gcctgcaggt taattaagat ctttttccct ctgccaaaaa ttatggggac
     5641 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     5701 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     5761 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
     5821 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     5881 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     5941 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     6001 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc
     6061 gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6121 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6181 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6241 aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta
     6301 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     6361 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     6421 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag
     6481 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6541 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc
     6601 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6661 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6721 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6781 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6841 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6901 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6961 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7021 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7081 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7141 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7201 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7261 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7321 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7381 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7441 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7501 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7561 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7621 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7681 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7741 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7801 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7861 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7921 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7981 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8041 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8101 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8161 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8221 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8281 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8341 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8401 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8461 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8521 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8581 gccacctgg
//
