LOCUS       sequence-381072-        7239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes multiple copies of an epitope tag OLLAS ( not detected by 
            anti GFP ab).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7239)
  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 7239)
  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_mRuby2_smFP OLLAS
FEATURES             Location/Qualifiers
     source          1..7239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(4..108)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        139..518
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          778..1794
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1718..1740
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1802..1821
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1916..1925
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1928..1969
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             1979..2020
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2030..2071
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2579..2620
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2630..2671
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2681..2722
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2741..2782
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2792..2833
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2843..2884
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             3077..3118
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             3128..3169
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             3179..3220
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     intron          3269..3401
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3384..3403
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3410..3998
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3463..3483)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3881..3892)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4025..4042)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4031..4142
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4238..4258)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4238..4256)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4277..4293)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4395..4414)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4460..4515
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4514..4533)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4876..4892)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4900..4916
                     /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(4924..4930,4931..4948,4949..4954))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4969..4990
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5049..5244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5095..5230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5250..5384
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5287..5306)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5341..5360
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5452..5469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5623..6211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5703..5722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6382..7173,7174..7239,1..3))
                     /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     7005..7024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
       61 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
      121 aaaagtgcca cctgggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg
      181 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      241 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      301 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      361 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      421 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      481 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
      541 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
      601 aattattttg tgcagcgatg ggggcggggg ggggggnnnn nnnnnggcga ggggcggggc
      661 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
      721 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
      781 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
      841 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
      901 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
      961 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg
     1021 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
     1081 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
     1141 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     1201 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     1261 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     1321 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1381 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1441 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1501 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1561 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1621 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1681 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1741 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1801 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgg taccgcgggc
     1861 ccggcgcgcc agctggatat ctccggaccc gggctagcgc aattgcggcc gcgccgccac
     1921 catggggtcc ggatttgcca atgaacttgg acctagactt atgggtaaac agcagcaatc
     1981 aggctttgcc aacgagttgg gccctcggtt gatgggaaag cagcagcagt ccgggttcgc
     2041 caacgaactg ggcccgaggc tcatggggaa gggccaggga aactcactga tcaaggaaaa
     2101 catgcggatg aaggtggtga tggagggctc cgtgaatggt caccagttca agtgcaccgg
     2161 agagggagag ggaaacccgt acatgggaac tcagaccatg cgcattaagg tcatcgaagg
     2221 aggtccgctg ccgttcgctt tcgatatcct ggccacttcg ttcggaggag ggtcgcgcac
     2281 gttcatcaag tacccgaagg gaatcccgga cttctttaag cagtcattcc cggaaggatt
     2341 cacttgggaa cgggtgaccc ggtatgaaga tggaggtgtg gtgactgtca tgcaagatac
     2401 ttcgctggag gatgggtgcc tcgtgtacca cgtccaagtc cgcggagtga atttcccgtc
     2461 caacggacca gtgatgcaga aaaagacgaa gggttgggaa cctaatactg aaatgatgta
     2521 ccccgcagac ggagggctga ggggctacac ccacatggcg ctgaaggtcg acggaggaag
     2581 cggatttgcg aatgagctcg gcccacgcct catgggcaaa caacagcaaa gcggttttgc
     2641 taacgaactg ggtccccgct tgatgggaaa gcagcagcag agcggattcg ctaatgagct
     2701 gggaccgagg cttatgggaa agggagggca gcagggcggt tccggtttcg cgaacgaatt
     2761 gggaccacgc ctgatgggga aacagcaaca gtccggcttc gccaatgagc ttgggccacg
     2821 ccttatgggg aagcagcaac aaagcgggtt cgctaacgag ctgggtcctc ggctgatggg
     2881 taaaggagga ggacatctgt cctgttcgtt cgtgaccacc tacagatcaa agaaaaccgt
     2941 gggaaacatc aagatgccgg gcattcatgc cgtcgaccac cgcctggagc ggctcgaaga
     3001 atcagacaat gagatgttcg tcgtgcaaag agaacatgcc gtggccaagt tcgcgggact
     3061 gggaggcggt caaggaagcg gctttgcaaa cgaactcggg ccaagattga tgggaaagca
     3121 acagcagtcg ggctttgcga acgagctcgg accaagactc atgggaaaac aacaacaatc
     3181 gggcttcgca aacgagcttg gacccagact gatgggcaaa tagaccggtg ctcgaggtac
     3241 cgatgaatag ctaaggtcga ggccgcaggt aagtatcaag gttacaagac aggtttaagg
     3301 agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     3361 tattggtctt actgacatcc actttgcctt tctctccaca ggtgtcgaca atcaacctct
     3421 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3481 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3541 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3601 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3661 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3721 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3781 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3841 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3901 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3961 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacga
     4021 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4081 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4141 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4201 ggggaggatt gggaagacaa tagcccagct tttgttccct ttagtgaggg ttaattgcgc
     4261 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctaattcact
     4321 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     4381 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     4441 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     4501 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     4561 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     4621 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     4681 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     4741 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     4801 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     4861 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4921 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4981 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5041 gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5101 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5161 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5221 tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca
     5281 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5341 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat
     5401 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5461 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5521 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5581 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5641 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5701 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5761 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5821 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5881 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5941 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6001 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6061 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6121 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6181 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6241 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6301 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6361 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6421 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6481 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6541 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6601 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6661 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6721 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6781 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6841 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6901 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6961 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7021 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7081 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7141 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7201 aaaaaaggga ataagggcga cacggaaatg ttgaatact
//
