LOCUS       Exported                6528 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Codon-optimized Archon2 fluorescent voltage reporter in zebrafish 
            expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    pTol2-UAS-zArchon2-KGC-EGFP-ER2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6528)
  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 6528)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6528
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    714..738
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(966..990)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      998..1007
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1871..2584
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     polyA_signal    3914..3995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3951..3970)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4005..4024
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4610..4627)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4781..5369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4861..4880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5540..6400)
                     /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     6163..6182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6401..6505)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtatctgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt ttactcaagt
       61 gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat ctggctagaa
      121 tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt attaaggaag
      181 taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa tctcaaaaaa
      241 cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta tttaattatc
      301 aaactataag acagacaatc taatgccagt acacgctact caaagttgta aaacctcaga
      361 tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc tcttttgggg
      421 ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc gggaagaggt
      481 gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga ctccctggtg
      541 tctgaaacac aggccagatg ggccattcgt acgcttaatt tcgaatatcg atatcaaggg
      601 cccatctcga gtatccggac acctaggtag catgcatggc gccttgtcga ctagctagca
      661 tcccgggaag aattcctcga cggatccacc ggtgtggaat tctgcagata tcaacaagtt
      721 tgtacaaaaa agcaggctgg ccatcaagct taggcctcca cggtggcttc taatccgtga
      781 gtcctagcgg gtgacagccc tccgtcttca caggcggagg agagtcttcc gtagggttcc
      841 tcggagtact gtcctccgac gcgtgcaagg gtcgactcta gagggtatat aatggatccc
      901 atcgcgtctc agcctcactt tgagctcctc cacacgaatt ccctcgacct cgaagacgcg
      961 tggataccca gctttcttgt acaaagtggt cactagtgcc accatggtga gcatcgctct
     1021 gcaggctgga tacgacctgc tgggagacgg aagacctgag acactgtggc tgggaatcgg
     1081 aacactgctg atgctgatcg gaacattcta cttcctggtg agaggatggg gagtgacaga
     1141 caaggacgct agagagtact acgctgtgcc tatcctggtg agcggaatcg ctagcgctgc
     1201 ttacctgagc atgttcttcg gaatcggact gacagaggtg cctgtgggag gagagatgct
     1261 ggacatctac tacgctagat acgctcactg gctgttcagc acacctctgc tgctgctgga
     1321 cctggctctg ctggctaagg tggacagagt gatcatcgga acactggtgg gagtggacgc
     1381 tctgatgatc gtgacaggac tgatcggagc tctgagccac acagctatcg ctagatacag
     1441 ctggtggctg ttcagcacaa tctgtatgat cgtggtgctg tacgtgctgg ctacaagcct
     1501 gagaagcgct gctaaggaga gaggacctga ggtggctagc acattcaaca tcctgacagc
     1561 tctggtgctg gtgctgtgga cagcttaccc tatcatctgg atcatcggaa cagagggagc
     1621 tggagtggtg ggactgggaa tcgagacact gctgttcatg gtgctggacg tgacatgtaa
     1681 ggtgggattc ggattcatcc tgctgagaag cagagctatc ctgggagaca cagaggctcc
     1741 tgagcctagc gctggagctg acagatctgc tgctgacaga cctgtggtgg ctgtgagcaa
     1801 ggctgctgct aagagcagaa tcacaagcga gggagagtac atccctctgg accagatcga
     1861 catcaacgtg gtgagcaagg gagaggagct gttcacagga gtggtgccta tcctggtgga
     1921 gctggacgga gacgtgaacg gacacaagtt cagcgtgagc ggagagggag agggagacgc
     1981 tacatacgga aagctgacac tgaagttcat ctgtacaaca ggaaagctgc ctgtgccttg
     2041 gcctacactg gtgacaacac tgacatacgg agtgcagtgt ttcagcagat accctgacca
     2101 catgaagcag cacgacttct tcaagagcgc tatgcctgag ggatacgtgc aggagagaac
     2161 aatcttcttc aaggacgacg gaaactacaa gacaagagct gaggtgaagt tcgagggaga
     2221 cacactggtg aacagaatcg agctgaaggg aatcgacttc aaggaggacg gaaacatcct
     2281 gggacacaag ctggagtaca actacaacag ccacaacgtg tacatcatgg ctgacaagca
     2341 gaagaacgga atcaaggtga acttcaagat cagacacaac atcgaggacg gaagcgtgca
     2401 gctggctgac cactaccagc agaacacacc tatcggagac ggacctgtgc tgctgcctga
     2461 caaccactac ctgagcacac agagcgctct gagcaaggac cctaacgaga agagagacca
     2521 catggtgctg ctggagttcg tgacagctgc tggaatcaca ctgggaatgg acgagctgta
     2581 caagttctgt tacgagaacg aggtgtgagg cgcgccaacc cgctcctcct ccctccctcc
     2641 ctccctccct cctctccttt attcactttt caacatgcac aacacgtcat caaatccccc
     2701 aatcagcact gaattacagc cacgggccac cagactgaag gtgttaaaga agacgattcg
     2761 gtctcatcag cggctaaaac gagatgaggg tgaaccgaga gagactgttt ccacagatgt
     2821 tcttgaagga gggccgggct cgtaggtgcg gttattacac atataagctc ccattcacac
     2881 cttaaaacat ccaacagcat tccaaagttt tccatcgggg accagggata gaaaacactg
     2941 cctcttccac tgcctaattt aacacattcc ttctagatta tctctggtca tctagatgaa
     3001 atcagttctg ttctgtttgc cgtgtctgaa aagacagtaa agagtaattg aaaaaaaaaa
     3061 tgaatataga aaaatgggga ctggagagat atttggacta gacgctcctg atccagccag
     3121 aaggaacctt tcaggtgtcg attacactgc ctgcgaaaac aacctcttct tccattcttc
     3181 agtttccagc tgccggctgg cttcttccac tgcccaacat cctaacaaac ccctgacaaa
     3241 aaaggagaca agccttgaaa catcctgaaa taagcaattc ttagctgcca gggacgtctt
     3301 cctgttagca tcagtgtatt tgtttctttt gctttgattt gttttctttt tttattttat
     3361 tttcatggta aatgtattct gcctgagcac tttaaacgaa ggtgtttttg ttccagtcga
     3421 tgcttcgttt tgaacctgct tttatattta ttaacttttg ttgtgtattt ttaatgttta
     3481 atctttaaag gaatagttca cctacacaca cacacacaca cacacacaca cacacaaatg
     3541 aaatgaagtc atcatttact cacttttctt tcttttagct gtggagcaca aaacgagatg
     3601 ttttgaagaa tgttcacgct gttgtttttc atgcaattta agtaaacgaa gatcaaggga
     3661 tgttgagcta tacaaatatt cattatatgg gcagcacggt cgctggttcg agtcccggct
     3721 gggccagttg gcatttctga gtttgcatat tctccctgtg ttggcatggg tttcctcctg
     3781 gtgctccgtt tctcccacag tccactagat cataatcagc cataccacat ttgtagaggt
     3841 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3901 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3961 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4021 catcaatgta tcaggcctta gggataacag ggtaatacgc gtgatctgcg aagatacggc
     4081 cacgggtgct cttgatcctg tggctgattt tggactgtgc tgctcgcagc tgctgatgaa
     4141 tcacatactt cctccatttt cttccactga ttgactgtta taatttccct aatttccagg
     4201 tcaaggtgct gtgcattgtg gtaatagatg tgacatgacg tcacttccaa aggaccaatg
     4261 aacatgtctg accaatttca tataatgtga aaacgatttt cataggcaga ataaataaca
     4321 tttaaattaa actgggcatc agcgcaattc aattggtttg gtaatagcaa gggaaaatag
     4381 aatgaagtga tctccaaaaa ataagtactt tttgactgta aataaaattg taaggagtaa
     4441 aaagtacttt tttttctaaa aaaatgtaat taagtaaaag taaaagtatt gatttttaat
     4501 tgtactcaag taaagtaaaa atccccaaaa ataatactta agtacagtaa tcaagtaaaa
     4561 ttactcaagt actttacacc tctggttctt gaggtacctc ttccgcttcc tcgctcactg
     4621 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4681 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4741 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4801 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4861 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4921 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4981 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5041 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5101 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5161 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5221 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5281 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5341 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5401 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5461 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5521 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5581 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5641 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5701 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5761 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5821 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5881 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5941 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6001 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6061 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6121 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6181 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6241 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6301 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6361 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6421 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6481 aaaataaaca aataggggtt ccgcgcacat ttccccgagg taccgatc
//
