LOCUS       sequence-211639-        6672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV production plasmid encoding for Archon1 fluorescent voltage 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6672)
  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 6672)
  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: pAAV-CaMKII-Archon1-KGC-EGFP
FEATURES             Location/Qualifiers
     source          1..6672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   35..164
                     /label=AAV2 ITR
     promoter        237..1525
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1536..1545
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2409..2411,2412..3125)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2409..2411,2412..3122)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2451..2472)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2712..2731)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3059..3080
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3150..3738
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3203..3223)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3621..3632)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3781..3902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3792..3811)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3846..3865
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4064..4193
                     /label=AAV2 ITR
     rep_origin      complement(4430..5018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4510..4529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5189..5980,5981..6049))
                     /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     5812..5831
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6050..6154)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6180..6635)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6317..6338)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6529..6548
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc
       61 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
      121 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa
      181 cccgccatgc tacttatcta cgtagccatg ctctaggaag atcgtaggtt aattaacatt
      241 atggccttag gtcacttcat ctccatgggg ttcttcttct gattttctag aaaatgagat
      301 gggggtgcag agagcttcct cagtgacctg cccagggtca catcagaaat gtcagagcta
      361 gaacttgaac tcagattact aatcttaaat tccatgcctt gggggcatgc aagtacgata
      421 tacagaagga gtgaactcat tagggcagat gaccaatgag tttaggaaag aagagtccag
      481 ggcagggtac atctacacca cccgcccagc cctgggtgag tccagccacg ttcacctcat
      541 tatagttgcc tctctccagt cctaccttga cgggaagcac aagcagaaac tgggacagga
      601 gccccaggag accaaatctt catggtccct ctgggaggat gggtggggag agctgtggca
      661 gaggcctcag gaggggccct gctgctcagt ggtgacagat aggggtgaga aagcagacag
      721 agtcattccg tcagcattct gggtctgttt ggtacttctt ctcacgctaa ggtggcggtg
      781 tgatatgcac aatggctaaa aagcagggag agctggaaag aaacaaggac agagacagag
      841 gccaagtcaa ccagaccaat tcccagagga agcaaagaaa ccattacaga gactacaagg
      901 gggaagggaa ggagagatga attagcttcc cctgtaaacc ttagaaccca gctgttgcca
      961 gggcaacggg gcaatacctg tctcttcaga ggagatgaag ttgccagggt aactacatcc
     1021 tgtctttctc aaggaccatc ccagaatgtg gcacccacta gccgttacca tagcaactgc
     1081 ctctttgccc cacttaatcc catcccgtct gttaaaaggg ccctatagtt ggaggtgggg
     1141 gaggtaggaa gagcgatgat cacttgtgga ctaagtttgt tcgcatcccc ttctccaacc
     1201 ccctcagtac atcaccctgg gggaacaggg tccacttgct cctgggccca cacagtcctg
     1261 cagtattgtg tatataaggc cagggcaaag aggagcaggt tttaaagtga aaggcaggca
     1321 ggtgttgggg aggcagttac cggggcaacg ggaacagggc gtttcggagg tggttgccat
     1381 ggggacctgg atgctgacga aggctcgcga ggctgtgagc agccacagtg ccctgctcag
     1441 aagccccaag ctcgtcagtc aagccggttc tccgtttgca ctcaggagca cgggcaggcg
     1501 agtggcccct agttctgggg gcagcggggg atcccgccac catggtttct atcgcactgc
     1561 aggctggtta tgacctcctc ggagatggta gacctgaaag tctgtggctc ggaattggta
     1621 cactgctgat gctgatcgga accttctact tcctggttag agcgtgggga gagactgaca
     1681 aggatgcacg ggagtactac gcagtgacaa tcctggtcag cggtattgcc agtgcagctt
     1741 atctgtcaat gttctttggt attggcctca cagaggtgcc agtaggtgga gagatgctga
     1801 acatctacta cgcaagatac gcacagtggc tgttcactac accactgttg ctgctgcacc
     1861 tggcactcct ggccaaagtt gatagagtga ccatcggcac tctcgttggc gtcgatgctc
     1921 tgatgattgt cacaggcctg ataggaacac tgtcacatac tgccattgcc agatactctt
     1981 ggtggctgtt ctcaactatc tgtatgatcg tcgttctgta tgtcctggcc acttctctgc
     2041 ggtcagccgc taaggagaga ggcccagagg ttgcctcaac cttcaacatc ctcaccgcac
     2101 tggttctggt cctgtggact gcttacccta tcatctggat tatcggcacc gagggtgccg
     2161 gagttgtggg cctgggaatc gaaactttgc tgttcatggt gctggacgtc accgccaagg
     2221 tgggcttcgg ttttatcctg ctcaggtcaa gagccattct gcaagataca gaagcacctg
     2281 aaccttcagc tggagctgat accggtgccg ccgaccgccc ggtagtagca gtgagcaagg
     2341 cggccgccaa gagcaggatc accagcgagg gcgagtacat ccccctggac cagatcgaca
     2401 tcaacgtggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2461 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2521 cctacggcaa gctgaccctg aagttcattt gcaccaccgg caagctgccc gtgccctggc
     2581 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     2641 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2701 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2761 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2821 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     2881 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     2941 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3001 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     3061 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3121 agtaagaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     3181 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3241 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3301 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3361 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3421 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3481 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3541 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3601 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3661 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3721 tttgggccgc ctccccgcat cgataccgtc gacccgggcg gccgcttcga gcagacatga
     3781 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3841 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3901 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     3961 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt cctagagcat
     4021 ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     4081 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     4141 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt
     4201 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4261 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4321 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4381 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4441 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4501 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4561 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4621 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4681 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4741 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4801 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4861 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4921 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4981 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5041 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5101 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5161 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5221 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5281 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5341 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5401 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5461 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5521 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5581 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5641 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5701 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5761 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5821 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5881 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5941 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6001 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6061 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6121 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     6181 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     6241 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     6301 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     6361 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     6421 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     6481 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     6541 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     6601 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     6661 ctgttgggaa gg
//
