LOCUS       sequence-208099-        6182 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Archon2 fluorescent voltage reporter in mammalian expression 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6182)
  AUTHORS   Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk HJ, Hochbaum
            DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang CT, Rhoades
            JL, Shemesh O, Asano S, Yoon YG, Freifeld L, Saulnier JL, Riegler C,
            Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, 
            Sabatini BL, Boyden ES
  TITLE     A robotic multidimensional directed evolution approach applied to 
            fluorescent voltage reporters.
  JOURNAL   Nat Chem Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9.
  PUBMED    29483642
REFERENCE   2  (bases 1 to 6182)
  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: "Nat Chem 
            Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9."
COMMENT     Sequence Label: pCAG-Archon2-KGC-GFP-ER2-WPRE
FEATURES             Location/Qualifiers
     source          1..6182
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     251..273
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     335..354
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      388..397
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1261..1263,1264..1974)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(1261..1263,1264..1974)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1303..1324)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1564..1583)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1911..1932
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          2035..2167
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2150..2169
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2176..2764
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2229..2249)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2647..2658)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2791..2808)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2797..2908
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3004..3024)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3004..3022)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3043..3059)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3165..3184)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3230..3285
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3284..3303)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3646..3662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3670..3686
                     /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(3694..3700,3701..3718,3719..3724))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3739..3760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3819..4014
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3865..4000
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4020..4154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4057..4076)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4111..4130
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4222..4239)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4393..4981)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4473..4492)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5152..5943,5944..6012))
                     /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     5775..5794
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6013..6117)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    6168..6179
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
ORIGIN
        1 tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg
       61 gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc
      121 ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc
      181 cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg
      241 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
      301 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
      361 catcattttg gcaaagaatt cggtaccgcc accatggttt ctatcgcact gcaggctggt
      421 tatgacctcc tcggagatgg tagacctgaa accctgtggc tcggaattgg tacactgctg
      481 atgctgatcg gaaccttcta cttcctggtt agagggtggg gagtgactga caaggatgca
      541 cgggagtact acgcagtgcc catcctggtc agcggtattg ccagtgcagc ttatctgtca
      601 atgttctttg gtatcggcct cacagaggtg ccagtcggtg gagagatgct ggacatctac
      661 tacgcaagat acgcacactg gctgttcagc acaccactgc tgctgctgga cctggcactc
      721 ctggccaaag ttgatagagt gatcatcggc actctcgttg gcgtcgatgc tctgatgatt
      781 gtcacaggcc tgataggagc actgtcacat actgccattg ccagatactc ttggtggctg
      841 ttctcaacta tctgtatgat cgtcgttctg tatgtcctgg ccacttctct gcggtcagcc
      901 gctaaggaga gaggcccaga ggttgcctca accttcaaca tcctcaccgc actggttctg
      961 gtcctgtgga ctgcttaccc tatcatctgg attatcggca ccgagggtgc cggagttgtg
     1021 ggcctgggaa tcgaaactct gctgttcatg gtgctggacg tcacctgcaa ggtgggcttc
     1081 gggtttatcc tgctcaggtc aagagccatt ctgggagata cagaagcacc tgaaccttca
     1141 gctggagctg ataccggtgc cgccgaccgc ccggtagtag cagtgagcaa ggcggccgcc
     1201 aagagcagga tcaccagcga gggcgagtac atccccctgg accagatcga catcaacgtg
     1261 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1321 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1381 aagctgaccc tgaagttcat ttgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1441 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     1501 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1561 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1621 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1681 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1741 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1801 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1861 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1921 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagttctgc
     1981 tacgagaacg aggtgtaatg tacaagtaaa gcggccccgg actcgaggcc gcaggtaagt
     2041 atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga
     2101 agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc
     2161 tccacaggtg tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2221 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2281 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2341 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2401 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2461 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2521 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     2581 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     2641 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     2701 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     2761 ccgcctggaa ttcgagctcg gtacgatcag cctcgactgt gccttctagt tgccagccat
     2821 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2881 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2941 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc ccagcttttg
     3001 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     3061 gtgaaattgt tatccgctaa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg
     3121 ctggtgtggc caatgccctg gctcacaaat accactgaga tcgatctttt tccctctgcc
     3181 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa
     3241 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     3301 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat
     3361 gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa
     3421 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     3481 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     3541 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     3601 ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
     3661 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3721 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3781 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
     3841 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3901 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3961 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
     4021 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4081 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4141 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4201 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4261 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4321 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4381 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4441 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4501 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4561 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4621 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4681 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4741 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4801 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4861 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4921 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4981 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5041 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5101 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5161 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5221 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5281 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5341 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5401 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5461 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5521 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5581 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5641 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5701 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5761 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5821 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5881 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5941 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6001 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6061 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6121 atttccccga aaagtgccac ctgggtcgac attgattatt gactagagtc atcctcatcg
     6181 tc
//
