LOCUS       sequence-345527-        8590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  B.1.617.2 variant S-protein with c-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8590)
  TITLE     Spike variants with FLAG tags
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8590)
  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            "Unpublished"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.1.617.2-FLAG
FEATURES             Location/Qualifiers
     source          1..8590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          660..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1688..1707
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             complement(3047..3064)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             5560..5583
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5611..5630)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5676..5731
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5730..5749)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6092..6108)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6092..6108)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6105..6127)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6116..6132
                     /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(6140..6146,6147..6164,6165..6170))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6185..6206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6265..6460
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6311..6446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6466..6600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6503..6522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6557..6576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6668..6685)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6839..7427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6919..6938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7598..8389,8390..8458))
                     /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     8221..8240
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8459..8563)
                     /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 gggsgcgcgc crgggggggg ggggsggggs ggggggsggg
      541 gsggggsgrg gcggrgaggk gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggrg ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcggatcc
     1741 atgtttgtgt tcctggtgct gctgccactg gtgtccagcc agtgtgtgaa cctgaggacc
     1801 aggacccaac ttcctcctgc ctacaccaac tccttcacca ggggagtcta ctaccctgac
     1861 aaggtgttca ggtcctctgt gctgcacagc acccaggacc tgttcctgcc attcttcagc
     1921 aatgtgacct ggttccatgc catccatgtg tctggcacca atggcaccaa gaggtttgac
     1981 aaccctgtgc tgccattcaa tgatggagtc tactttgcca gcacagagaa gagcaacatc
     2041 atcaggggct ggatttttgg caccaccctg gacagcaaga cccagtccct gctgattgtg
     2101 aacaatgcca ccaatgtggt gattaaggtg tgtgagttcc agttctgtaa tgacccattc
     2161 ctgggagtct actaccacaa gaacaacaag tcctggatgg agtctggggt ctactcctct
     2221 gccaacaact gtacctttga atatgtgagc caaccattcc tgatggactt ggagggcaag
     2281 cagggcaact tcaagaacct gagggagttt gtgttcaaga acattgatgg ctacttcaag
     2341 atttacagca aacacacacc aatcaacctg gtgagggacc tgccacaggg cttctctgcc
     2401 ttggaaccac tggtggacct gccaattggc atcaacatca ccaggttcca gaccctgctg
     2461 gctctgcaca ggtcctacct gacacctgga gactcctcct ctggctggac agcaggagca
     2521 gcagcctact atgtgggcta cctccaacca aggaccttcc tgctgaaata caatgagaat
     2581 ggcaccatca cagatgctgt ggactgtgcc ctggacccac tgtctgagac caagtgtacc
     2641 ctgaaatcct tcacagtgga gaagggcatc taccagacca gcaacttcag ggtccaacca
     2701 acagagagca ttgtgaggtt tccaaacatc accaacctgt gtccatttgg agaggtgttc
     2761 aatgccacca ggtttgcctc tgtctatgcc tggaacagga agaggattag caactgtgtg
     2821 gctgactact ctgtgctcta caactctgcc tccttcagca ccttcaagtg ttatggagtg
     2881 agcccaacca aactgaatga cctgtgtttc accaatgtct atgctgactc ctttgtgatt
     2941 aggggagatg aggtgagaca gattgcccct ggacaaacag gcaagattgc tgactacaac
     3001 tacaaactgc ctgatgactt cacaggctgt gtgattgcct ggaacagcaa caacctggac
     3061 agcaaggtgg gaggcaacta caactacagg tacagactgt tcaggaagag caacctgaaa
     3121 ccatttgaga gggacatcag cacagagatt taccaggctg gcagcaaacc atgtaatgga
     3181 gtggagggct tcaactgtta ctttccactc caatcctatg gcttccaacc aaccaatgga
     3241 gtgggctacc aaccatacag ggtggtggtg ctgtcctttg aactgctcca tgcccctgcc
     3301 acagtgtgtg gaccaaagaa gagcaccaac ctggtgaaga acaagtgtgt gaacttcaac
     3361 ttcaatggac tgacaggcac aggagtgctg acagagagca acaagaagtt cctgccattc
     3421 caacagtttg gcagggacat tgctgacacc acagatgctg tgagggaccc acagaccttg
     3481 gagattctgg acatcacacc atgttccttt ggaggagtgt ctgtgattac acctggcacc
     3541 aacaccagca accaggtggc tgtgctctac cagggtgtga actgtactga ggtgcctgtg
     3601 gctatccatg ctgaccaact tacaccaacc tggagggtct acagcacagg cagcaatgtg
     3661 ttccagacca gggctggctg tctgattgga gcagagcatg tgaacaactc ctatgagtgt
     3721 gacatcccaa ttggagcagg catctgtgcc tcctaccaga cccagaccaa cagcagaagg
     3781 agggcaaggt ctgtggcaag ccagagcatc attgcctaca caatgagtct gggagcagag
     3841 aactctgtgg cttacagcaa caacagcatt gccatcccaa ccaacttcac catctctgtg
     3901 accacagaga ttctgcctgt gagtatgacc aagacctctg tggactgtac aatgtatatc
     3961 tgtggagaca gcacagagtg tagcaacctg ctgctccaat atggctcctt ctgtacccaa
     4021 cttaacaggg ctctgacagg cattgctgtg gaacaggaca agaacaccca ggaggtgttt
     4081 gcccaggtga agcagattta caagacacct ccaatcaagg actttggagg cttcaacttc
     4141 agccagattc tgcctgaccc aagcaagcca agcaagaggt ccttcattga ggacctgctg
     4201 ttcaacaagg tgaccctggc tgatgctggc ttcatcaagc aatatggaga ctgtctggga
     4261 gacattgctg ccagggacct gatttgtgcc cagaagttca atggactgac agtgctgcct
     4321 ccactgctga cagatgagat gattgcccaa tacacctctg ccctgctggc tggcaccatc
     4381 acctctggct ggacctttgg agcaggagca gccctccaaa tcccatttgc tatgcagatg
     4441 gcttacaggt tcaatggcat tggagtgacc cagaatgtgc tctatgagaa ccagaaactg
     4501 attgccaacc agttcaactc tgccattggc aagattcagg actccctgtc cagcacagcc
     4561 tctgccctgg gcaaactcca aaatgtggtg aaccagaatg cccaggctct gaacaccctg
     4621 gtgaagcaac tttccagcaa ctttggagcc atctcctctg tgctgaatga catcctgagc
     4681 agactggaca aggtggaggc tgaggtccag attgacagac tgattacagg cagactccaa
     4741 tccctccaaa cctatgtgac ccaacaactt atcagggctg ctgagattag ggcatctgcc
     4801 aacctggctg ccaccaagat gagtgagtgt gtgctgggac aaagcaagag ggtggacttc
     4861 tgtggcaagg gctaccacct gatgagtttt ccacagtctg cccctcatgg agtggtgttc
     4921 ctgcatgtga cctatgtgcc tgcccaggag aagaacttca ccacagcccc tgccatctgc
     4981 catgatggca aggctcactt tccaagggag ggagtgtttg tgagcaatgg cacccactgg
     5041 tttgtgaccc agaggaactt ctatgaacca cagattatca ccacagacaa cacctttgtg
     5101 tctggcaact gtgatgtggt gattggcatt gtgaacaaca cagtctatga cccactccaa
     5161 cctgaactgg actccttcaa ggaggaactg gacaaatact tcaagaacca caccagccct
     5221 gatgtggacc tgggagacat ctctggcatc aatgcctctg tggtgaacat ccagaaggag
     5281 attgacagac tgaatgaggt ggctaagaac ctgaatgagt ccctgattga cctccaagaa
     5341 ctgggcaaat atgaacaata catcaagtgg ccatggtaca tctggctggg cttcattgct
     5401 ggactgattg ccattgtgat ggtcaccatc atgctgtgct gcatgacctc atgttgctcc
     5461 tgtcttaagg ggtgctgctc ctgcggttcc tgttgcaaat ttgacgagga cgacagcgaa
     5521 ccggtgctta agggtgtgaa gctgcattac accggaggcg attacaagga tgacgatgac
     5581 aagtgacggc cgcctgcagg ttaattaaga tctttttccc tctgccaaaa attatgggga
     5641 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     5701 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     5761 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     5821 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     5881 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     5941 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     6001 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     6061 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6121 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6181 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6241 gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct
     6301 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6361 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6421 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca
     6481 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6541 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     6601 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6661 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6721 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6781 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6841 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6901 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6961 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7021 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7081 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7141 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7201 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7261 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7321 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7381 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7441 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7501 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7561 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7621 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7681 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7741 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7801 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7861 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7921 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7981 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     8041 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8101 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     8161 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8221 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8281 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8341 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8401 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8461 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8521 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8581 tgccacctgg
//
