LOCUS       sequence-204582-        6518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Codon-optimized Archon2 fluorescent voltage reporter in zebrafish 
            expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6518)
  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 6518)
  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: pTol2-UAS-zArchon2-KGC-EGFP-ER2
FEATURES             Location/Qualifiers
     source          1..6518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(18..29,30..192)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    708..732
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(960..984)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      992..1001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1865..1867,1868..2578)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     polyA_signal    3908..3989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3945..3964)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3999..4018
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(4379..4566,4567..4578)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4604..4621)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4775..5363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4855..4874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5534..6325,6326..6394))
                     /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     6157..6176
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6395..6499)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgtccaggag ttcttgacag aggtgtaaaa agtactcaaa aattttactc aagtgaaagt
       61 acaagtactt agggaaaatt ttactcaatt aaaagtaaaa gtatctggct agaatcttac
      121 ttgagtaaaa gtaaaaaagt actccattaa aattgtactt gagtattaag gaagtaaaag
      181 taaaagcaag aaagaaaact agagattctt gtttaagctt ttaatctcaa aaaacattaa
      241 atgaaatgca tacaaggttt tatcctgctt tagaactgtt tgtatttaat tatcaaacta
      301 taagacagac aatctaatgc cagtacacgc tactcaaagt tgtaaaacct cagatttaac
      361 ttcagtagaa gctgattctc aaaattgtta gtgtcaagcc tagctctttt ggggctgaaa
      421 agcaatcctg cagtgctgaa aagcctctca caggcagccg atgcgggaag aggtgtatta
      481 gtcttgatag agaggctgca aatagcagga aacgtgagca gagactccct ggtgtctgaa
      541 acacaggcca gatgggccat tcgtacgctt aatttcgaat atcgatatca agggcccatc
      601 tcgagtatcc ggacacctag gtagcatgca tggcgccttg tcgactagct agcatcccgg
      661 gaagaattcc tcgacggatc caccggtgtg gaattctgca gatatcaaca agtttgtaca
      721 aaaaagcagg ctggccatca agcttaggcc tccacggtgg cttctaatcc gtgagtccta
      781 gcgggtgaca gccctccgtc ttcacaggcg gaggagagtc ttccgtaggg ttcctcggag
      841 tactgtcctc cgacgcgtgc aagggtcgac tctagagggt atataatgga tcccatcgcg
      901 tctcagcctc actttgagct cctccacacg aattccctcg acctcgaaga cgcgtggata
      961 cccagctttc ttgtacaaag tggtcactag tgccaccatg gtgagcatcg ctctgcaggc
     1021 tggatacgac ctgctgggag acggaagacc tgagacactg tggctgggaa tcggaacact
     1081 gctgatgctg atcggaacat tctacttcct ggtgagagga tggggagtga cagacaagga
     1141 cgctagagag tactacgctg tgcctatcct ggtgagcgga atcgctagcg ctgcttacct
     1201 gagcatgttc ttcggaatcg gactgacaga ggtgcctgtg ggaggagaga tgctggacat
     1261 ctactacgct agatacgctc actggctgtt cagcacacct ctgctgctgc tggacctggc
     1321 tctgctggct aaggtggaca gagtgatcat cggaacactg gtgggagtgg acgctctgat
     1381 gatcgtgaca ggactgatcg gagctctgag ccacacagct atcgctagat acagctggtg
     1441 gctgttcagc acaatctgta tgatcgtggt gctgtacgtg ctggctacaa gcctgagaag
     1501 cgctgctaag gagagaggac ctgaggtggc tagcacattc aacatcctga cagctctggt
     1561 gctggtgctg tggacagctt accctatcat ctggatcatc ggaacagagg gagctggagt
     1621 ggtgggactg ggaatcgaga cactgctgtt catggtgctg gacgtgacat gtaaggtggg
     1681 attcggattc atcctgctga gaagcagagc tatcctggga gacacagagg ctcctgagcc
     1741 tagcgctgga gctgacagat ctgctgctga cagacctgtg gtggctgtga gcaaggctgc
     1801 tgctaagagc agaatcacaa gcgagggaga gtacatccct ctggaccaga tcgacatcaa
     1861 cgtggtgagc aagggagagg agctgttcac aggagtggtg cctatcctgg tggagctgga
     1921 cggagacgtg aacggacaca agttcagcgt gagcggagag ggagagggag acgctacata
     1981 cggaaagctg acactgaagt tcatctgtac aacaggaaag ctgcctgtgc cttggcctac
     2041 actggtgaca acactgacat acggagtgca gtgtttcagc agataccctg accacatgaa
     2101 gcagcacgac ttcttcaaga gcgctatgcc tgagggatac gtgcaggaga gaacaatctt
     2161 cttcaaggac gacggaaact acaagacaag agctgaggtg aagttcgagg gagacacact
     2221 ggtgaacaga atcgagctga agggaatcga cttcaaggag gacggaaaca tcctgggaca
     2281 caagctggag tacaactaca acagccacaa cgtgtacatc atggctgaca agcagaagaa
     2341 cggaatcaag gtgaacttca agatcagaca caacatcgag gacggaagcg tgcagctggc
     2401 tgaccactac cagcagaaca cacctatcgg agacggacct gtgctgctgc ctgacaacca
     2461 ctacctgagc acacagagcg ctctgagcaa ggaccctaac gagaagagag accacatggt
     2521 gctgctggag ttcgtgacag ctgctggaat cacactggga atggacgagc tgtacaagtt
     2581 ctgttacgag aacgaggtgt gaggcgcgcc aacccgctcc tcctccctcc ctccctccct
     2641 ccctcctctc ctttattcac ttttcaacat gcacaacacg tcatcaaatc ccccaatcag
     2701 cactgaatta cagccacggg ccaccagact gaaggtgtta aagaagacga ttcggtctca
     2761 tcagcggcta aaacgagatg agggtgaacc gagagagact gtttccacag atgttcttga
     2821 aggagggccg ggctcgtagg tgcggttatt acacatataa gctcccattc acaccttaaa
     2881 acatccaaca gcattccaaa gttttccatc ggggaccagg gatagaaaac actgcctctt
     2941 ccactgccta atttaacaca ttccttctag attatctctg gtcatctaga tgaaatcagt
     3001 tctgttctgt ttgccgtgtc tgaaaagaca gtaaagagta attgaaaaaa aaaatgaata
     3061 tagaaaaatg gggactggag agatatttgg actagacgct cctgatccag ccagaaggaa
     3121 cctttcaggt gtcgattaca ctgcctgcga aaacaacctc ttcttccatt cttcagtttc
     3181 cagctgccgg ctggcttctt ccactgccca acatcctaac aaacccctga caaaaaagga
     3241 gacaagcctt gaaacatcct gaaataagca attcttagct gccagggacg tcttcctgtt
     3301 agcatcagtg tatttgtttc ttttgctttg atttgttttc tttttttatt ttattttcat
     3361 ggtaaatgta ttctgcctga gcactttaaa cgaaggtgtt tttgttccag tcgatgcttc
     3421 gttttgaacc tgcttttata tttattaact tttgttgtgt atttttaatg tttaatcttt
     3481 aaaggaatag ttcacctaca cacacacaca cacacacaca cacacacaca aatgaaatga
     3541 agtcatcatt tactcacttt tctttctttt agctgtggag cacaaaacga gatgttttga
     3601 agaatgttca cgctgttgtt tttcatgcaa tttaagtaaa cgaagatcaa gggatgttga
     3661 gctatacaaa tattcattat atgggcagca cggtcgctgg ttcgagtccc ggctgggcca
     3721 gttggcattt ctgagtttgc atattctccc tgtgttggca tgggtttcct cctggtgctc
     3781 cgtttctccc acagtccact agatcataat cagccatacc acatttgtag aggttttact
     3841 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3901 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3961 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4021 tgtatcaggc cttagggata acagggtaat acgcgtgatc tgcgaagata cggccacggg
     4081 tgctcttgat cctgtggctg attttggact gtgctgctcg cagctgctga tgaatcacat
     4141 acttcctcca ttttcttcca ctgattgact gttataattt ccctaatttc caggtcaagg
     4201 tgctgtgcat tgtggtaata gatgtgacat gacgtcactt ccaaaggacc aatgaacatg
     4261 tctgaccaat ttcatataat gtgaaaacga ttttcatagg cagaataaat aacatttaaa
     4321 ttaaactggg catcagcgca attcaattgg tttggtaata gcaagggaaa atagaatgaa
     4381 gtgatctcca aaaaataagt actttttgac tgtaaataaa attgtaagga gtaaaaagta
     4441 cttttttttc taaaaaaatg taattaagta aaagtaaaag tattgatttt taattgtact
     4501 caagtaaagt aaaaatcccc aaaaataata cttaagtaca gtaatcaagt aaaattactc
     4561 aagtacttta cacctctggt tcttgaggta cctcttccgc ttcctcgctc actgactcgc
     4621 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4681 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4741 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4801 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4861 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4921 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4981 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5041 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5101 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5161 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5221 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5281 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5341 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5401 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5461 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5521 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5581 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5641 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5701 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5761 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5821 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5881 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5941 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6001 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6061 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6121 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6181 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6241 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6301 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6361 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6421 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6481 aacaaatagg ggttccgcgc acatttcccc gaggtacc
//
