LOCUS       sequence-345528-        8598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P1 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  JOURNAL   Unpublished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            "Unpublished"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-P1-FLAG
FEATURES             Location/Qualifiers
     source          1..8598
                     /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(3055..3072)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             5568..5591
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5619..5638)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5684..5739
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5738..5757)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6100..6116)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6100..6116)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6113..6135)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6124..6140
                     /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(6148..6154,6155..6172,6173..6178))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6193..6214
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6273..6468
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6319..6454
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6474..6608
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6511..6530)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6565..6584
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6676..6693)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6847..7435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6927..6946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7606..8397,8398..8466))
                     /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     8229..8248
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8467..8571)
                     /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 rggsggrgag gkgcggcggc 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 ggaggggcgc 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 aacttcacca
     1801 acaggaccca acttccttct gcctacacca actccttcac caggggagtc tactaccctg
     1861 acaaggtgtt caggtcctct gtgctgcaca gcacccagga cctgttcctg ccattcttca
     1921 gcaatgtgac ctggttccat gccatccatg tgtctggcac caatggcacc aagaggtttg
     1981 acaaccctgt gctgccattc aatgatggag tctactttgc cagcacagag aagagcaaca
     2041 tcatcagggg ctggattttt ggcaccaccc tggacagcaa gacccagtcc ctgctgattg
     2101 tgaacaatgc caccaatgtg gtgattaagg tgtgtgagtt ccagttctgt aattacccat
     2161 tcctgggagt ctactaccac aagaacaaca agtcctggat ggagtctgag ttcagggtct
     2221 actcctctgc caacaactgt acctttgaat atgtgagcca accattcctg atggacttgg
     2281 agggcaagca gggcaacttc aagaacctgt cggagtttgt gttcaagaac attgatggct
     2341 acttcaagat ttacagcaaa cacacaccaa tcaacctggt gagggacctg ccacagggct
     2401 tctctgcctt ggaaccactg gtggacctgc caattggcat caacatcacc aggttccaga
     2461 ccctgctggc tctgcacagg tcctacctga cacctggaga ctcctcctct ggctggacag
     2521 caggagcagc agcctactat gtgggctacc tccaaccaag gaccttcctg ctgaaataca
     2581 atgagaatgg caccatcaca gatgctgtgg actgtgccct ggacccactg tctgagacca
     2641 agtgtaccct gaaatccttc acagtggaga agggcatcta ccagaccagc aacttcaggg
     2701 tccaaccaac agagagcatt gtgaggtttc caaacatcac caacctgtgt ccatttggag
     2761 aggtgttcaa tgccaccagg tttgcctctg tctatgcctg gaacaggaag aggattagca
     2821 actgtgtggc tgactactct gtgctctaca actctgcctc cttcagcacc ttcaagtgtt
     2881 atggagtgag cccaaccaaa ctgaatgacc tgtgtttcac caatgtctat gctgactcct
     2941 ttgtgattag gggagatgag gtgagacaga ttgcccctgg acaaacaggc acgattgctg
     3001 actacaacta caaactgcct gatgacttca caggctgtgt gattgcctgg aacagcaaca
     3061 acctggacag caaggtggga ggcaactaca actacctcta cagactgttc aggaagagca
     3121 acctgaaacc atttgagagg gacatcagca cagagattta ccaggctggc agcacaccat
     3181 gtaatggagt gaagggcttc aactgttact ttccactcca atcctatggc ttccaaccaa
     3241 cctatggagt gggctaccaa ccatacaggg tggtggtgct gtcctttgaa ctgctccatg
     3301 cccctgccac agtgtgtgga ccaaagaaga gcaccaacct ggtgaagaac aagtgtgtga
     3361 acttcaactt caatggactg acaggcacag gagtgctgac agagagcaac aagaagttcc
     3421 tgccattcca acagtttggc agggacattg ctgacaccac agatgctgtg agggacccac
     3481 agaccttgga gattctggac atcacaccat gttcctttgg aggagtgtct gtgattacac
     3541 ctggcaccaa caccagcaac caggtggctg tgctctacca gggtgtgaac tgtactgagg
     3601 tgcctgtggc tatccatgct gaccaactta caccaacctg gagggtctac agcacaggca
     3661 gcaatgtgtt ccagaccagg gctggctgtc tgattggagc agagtatgtg aacaactcct
     3721 atgagtgtga catcccaatt ggagcaggca tctgtgcctc ctaccagacc cagaccaaca
     3781 gcccaaggag ggcaaggtct gtggcaagcc agagcatcat tgcctacaca atgagtctgg
     3841 gagcagagaa ctctgtggct tacagcaaca acagcattgc catcccaacc aacttcacca
     3901 tctctgtgac cacagagatt ctgcctgtga gtatgaccaa gacctctgtg gactgtacaa
     3961 tgtatatctg tggagacagc acagagtgta gcaacctgct gctccaatat ggctccttct
     4021 gtacccaact taacagggct ctgacaggca ttgctgtgga acaggacaag aacacccagg
     4081 aggtgtttgc ccaggtgaag cagatttaca agacacctcc aatcaaggac tttggaggct
     4141 tcaacttcag ccagattctg cctgacccaa gcaagccaag caagaggtcc ttcattgagg
     4201 acctgctgtt caacaaggtg accctggctg atgctggctt catcaagcaa tatggagact
     4261 gtctgggaga cattgctgcc agggacctga tttgtgccca gaagttcaat ggactgacag
     4321 tgctgcctcc actgctgaca gatgagatga ttgcccaata cacctctgcc ctgctggctg
     4381 gcaccatcac ctctggctgg acctttggag caggagcagc cctccaaatc ccatttgcta
     4441 tgcagatggc ttacaggttc aatggcattg gagtgaccca gaatgtgctc tatgagaacc
     4501 agaaactgat tgccaaccag ttcaactctg ccattggcaa gattcaggac tccctgtcca
     4561 gcacagcctc tgccctgggc aaactccaag atgtggtgaa ccagaatgcc caggctctga
     4621 acaccctggt gaagcaactt tccagcaact ttggagccat ctcctctgtg ctgaatgaca
     4681 tcctgagcag actggacaag gtggaggctg aggtccagat tgacagactg attacaggca
     4741 gactccaatc cctccaaacc tatgtgaccc aacaacttat cagggctgct gagattaggg
     4801 catctgccaa cctggctgcc atcaagatga gtgagtgtgt gctgggacaa agcaagaggg
     4861 tggacttctg tggcaagggc taccacctga tgagttttcc acagtctgcc cctcatggag
     4921 tggtgttcct gcatgtgacc tatgtgcctg cccaggagaa gaacttcacc acagcccctg
     4981 ccatctgcca tgatggcaag gctcactttc caagggaggg agtgtttgtg agcaatggca
     5041 cccactggtt tgtgacccag aggaacttct atgaaccaca gattatcacc acagacaaca
     5101 cctttgtgtc tggcaactgt gatgtggtga ttggcattgt gaacaacaca gtctatgacc
     5161 cactccaacc tgaactggac tccttcaagg aggaactgga caaatacttc aagaaccaca
     5221 ccagccctga tgtggacctg ggagacatct ctggcatcaa tgcctctgtg gtgaacatcc
     5281 agaaggagat tgacagactg aatgaggtgg ctaagaacct gaatgagtcc ctgattgacc
     5341 tccaagaact gggcaaatat gaacaataca tcaagtggcc atggtacatc tggctgggct
     5401 tcattgctgg actgattgcc attgtgatgg tgaccatcat gctgtgctgc atgacctcat
     5461 gttgctcctg tcttaagggg tgctgctcct gcggttcctg ttgcaaattt gacgaggacg
     5521 acagcgaacc ggtgcttaag ggtgtgaagc tgcattacac cggaggcgat tacaaggatg
     5581 acgatgacaa gtgacggccg cctgcaggtt aattaagatc tttttccctc tgccaaaaat
     5641 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     5701 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     5761 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     5821 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     5881 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     5941 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     6001 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     6061 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6121 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6181 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6241 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     6301 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6361 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     6421 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     6481 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6541 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6601 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6661 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6721 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6781 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6841 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6901 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6961 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7021 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7081 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7141 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7201 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7261 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7321 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7381 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7441 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7501 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7561 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7621 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7681 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7741 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7801 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7861 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7921 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7981 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8041 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8101 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8161 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8221 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8281 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8341 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8401 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8461 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8521 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8581 ccgaaaagtg ccacctgg
//
