LOCUS       sequence-457329-        7168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes multiple copies of an epitope tag FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7168)
  AUTHORS   Viswanathan S, Williams ME, Bloss EB, Stasevich TJ, Speer CM, Nern 
            A, Pfeiffer BD, Hooks BM, Li WP, English BP, Tian T, Henry GL, 
            Macklin JJ, Patel R, Gerfen CR, Zhuang X, Wang Y, Rubin GM, Looger 
            LL
  TITLE     High-performance probes for light and electron microscopy.
  JOURNAL   Nat Methods. 2015 Jun;12(6):568-76. doi: 10.1038/nmeth.3365. Epub 
            2015 Apr 27.
  PUBMED    25915120
REFERENCE   2  (bases 1 to 7168)
  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"; doi: 
            "10.1038/nmeth.3365"; journalName: "Nat Methods"; date: "2015-06"; 
            volume: "12"; issue: "6"; pages: "568-76"
COMMENT     Sequence Label: pCAG_smFP FLAG
FEATURES             Location/Qualifiers
     source          1..7168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          227..1243
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1167..1189
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1251..1270
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1360..1369
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1369..1392
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1396..1419
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1423..1446
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1975..1998
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2005..2028
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2041..2064
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2071..2094
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2305..2328
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2332..2355
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2359..2382
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     intron          2431..2563
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2546..2565
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2572..3160
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2625..2645)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3043..3054)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3187..3204)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3193..3304
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3400..3420)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3400..3418)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3439..3455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3557..3576)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3622..3677
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3676..3695)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4038..4054)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4062..4078
                     /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(4086..4092,4093..4110,4111..4116))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4131..4152
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4211..4406
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4257..4392
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4412..4546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4449..4468)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4503..4522
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4614..4631)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4785..5373)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4865..4884)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5544..6335,6336..6404))
                     /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     6167..6186
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6405..6509)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6540..6919
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cccctgaagt tggtcatttt ttgcgggggg gggggggggg ggaggggcgg gggggggggg
       61 gggggggggg ggggcggggg gggggggggg gcgggggggg ggggggggcg gggcgaggcg
      121 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
      181 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
      241 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
      301 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
      361 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
      421 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
      481 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc
      541 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg
      601 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg
      661 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt
      721 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc
      781 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc
      841 cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc
      901 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt
      961 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg
     1021 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc
     1081 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg
     1141 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag
     1201 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     1261 ggttattgtg ctgtctcatc attttggcaa agaattcggt accgcgggcc cggcgcgcca
     1321 gctggatatc tccggacccg ggctagcgca attgcggccg ccaccatgga ctacaaggac
     1381 gacgacgata aaggagatta caaagatgac gatgataagg gcgattataa ggacgacgat
     1441 gacaagggcg gcgtgtcgaa aggcgaggag ctgttcacag gcgtggtccc catcctggtt
     1501 gagctggacg gtgacgttaa cggccacaaa tttagcgtac gtggtgaggg cgagggtgac
     1561 gctaccaacg gaaagctgac cctgaaattc atctgcacaa ctggcaagct gccggtgccc
     1621 tggcccacgc tggtaaccac tctgggcgga ggcgttcaat gtttcagtcg ttacccggat
     1681 cacatgaagc agcatgactt cttcaagtcg gccatgcccg aaggctacgt gcaggagcgc
     1741 accatcagtt ttaaggatga cggcacctac aagactcgcg ccgaggtcaa gttcgaagga
     1801 gacaccctgg ttaatcgtat cgagctcaaa ggcatcgatt tcaaggagga cggaaatatt
     1861 ctgggccaca agctggagta caattttaac agccacaatg tttacattac tgccgacaag
     1921 cagaagaacg gcatcaaggc taatttcaag attcgtcaca atgtggaggg aggcgactat
     1981 aaggatgacg atgacaaaca acaagactat aaggacgatg atgacaaagg tcagcaaggc
     2041 gattacaagg acgatgacga taagcagcaa gattacaagg atgatgatga taagggaggc
     2101 gacggaagcg tgcagctggc agatcattac caacagaaca cgcccattgg tgacggtcca
     2161 gtcctgctgc ctgataacca ctacttgtcc acccagtcgg tgctgtcgaa ggacccgaac
     2221 gagaagcgtg accacatggt gctgctggag ttcgtgaccg ccgcaggaat caccctcggc
     2281 atggatgagc tttacaaggg cggagattat aaagatgacg acgacaaggg cgactacaaa
     2341 gatgatgatg acaagggtga ctacaaagac gacgatgata aataaaccgg tgctcgaggt
     2401 accgatgaat agctaaggtc gaggccgcag gtaagtatca aggttacaag acaggtttaa
     2461 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
     2521 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcga caatcaacct
     2581 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2641 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2701 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2761 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2821 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2881 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2941 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     3001 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3061 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3121 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattcg agctcggtac
     3181 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3241 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3301 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3361 agggggagga ttgggaagac aatagcccag cttttgttcc ctttagtgag ggttaattgc
     3421 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctaattca
     3481 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
     3541 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     3601 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     3661 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     3721 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     3781 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     3841 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     3901 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     3961 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca
     4021 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4081 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4141 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4201 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4261 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4321 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4381 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     4441 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4501 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     4561 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4621 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4681 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4741 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4801 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4861 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4921 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4981 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5041 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5101 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5161 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5221 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5281 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5341 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5401 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5461 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5521 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5581 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5641 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5701 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5761 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5821 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5881 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5941 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6001 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6061 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6121 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6181 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6241 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6301 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6361 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6421 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6481 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcg
     6541 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     6601 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     6661 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     6721 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     6781 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     6841 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6901 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
     6961 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
     7021 gggggcgggg gggggggggg ggcgcgcccc gggggggggg gggggggggg gggggggggg
     7081 gggggggggg gggggggggg ggggcccggg cgggcggggg ggggggggcc ggagggtttc
     7141 ttttttttgg tgcgggcggg ggggtcgg
//
