LOCUS       sequence-260833-        6692 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 6692)
  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 6692)
  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-ER2
FEATURES             Location/Qualifiers
     source          1..6692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..1312
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1322..1331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2195..2197,2198..2908)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2195..2197,2198..2908)
                     /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(2237..2258)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2498..2517)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2845..2866
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2957..3545
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3010..3030)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3428..3439)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3588..3709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3599..3618)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3653..3672
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3871..4000
                     /label=AAV2 ITR
     rep_origin      complement(4237..4825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4317..4336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4996..5787,5788..5856))
                     /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     5619..5638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5857..5961)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5987..6442)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6124..6145)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6336..6355
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6514..6643
                     /label=AAV2 ITR
ORIGIN
        1 taggaagatc gtaggttaat taacattatg gccttaggtc acttcatctc catggggttc
       61 ttcttctgat tttctagaaa atgagatggg ggtgcagaga gcttcctcag tgacctgccc
      121 agggtcacat cagaaatgtc agagctagaa cttgaactca gattactaat cttaaattcc
      181 atgccttggg ggcatgcaag tacgatatac agaaggagtg aactcattag ggcagatgac
      241 caatgagttt aggaaagaag agtccagggc agggtacatc tacaccaccc gcccagccct
      301 gggtgagtcc agccacgttc acctcattat agttgcctct ctccagtcct accttgacgg
      361 gaagcacaag cagaaactgg gacaggagcc ccaggagacc aaatcttcat ggtccctctg
      421 ggaggatggg tggggagagc tgtggcagag gcctcaggag gggccctgct gctcagtggt
      481 gacagatagg ggtgagaaag cagacagagt cattccgtca gcattctggg tctgtttggt
      541 acttcttctc acgctaaggt ggcggtgtga tatgcacaat ggctaaaaag cagggagagc
      601 tggaaagaaa caaggacaga gacagaggcc aagtcaacca gaccaattcc cagaggaagc
      661 aaagaaacca ttacagagac tacaaggggg aagggaagga gagatgaatt agcttcccct
      721 gtaaacctta gaacccagct gttgccaggg caacggggca atacctgtct cttcagagga
      781 gatgaagttg ccagggtaac tacatcctgt ctttctcaag gaccatccca gaatgtggca
      841 cccactagcc gttaccatag caactgcctc tttgccccac ttaatcccat cccgtctgtt
      901 aaaagggccc tatagttgga ggtgggggag gtaggaagag cgatgatcac ttgtggacta
      961 agtttgttcg catccccttc tccaaccccc tcagtacatc accctggggg aacagggtcc
     1021 acttgctcct gggcccacac agtcctgcag tattgtgtat ataaggccag ggcaaagagg
     1081 agcaggtttt aaagtgaaag gcaggcaggt gttggggagg cagttaccgg ggcaacggga
     1141 acagggcgtt tcggaggtgg ttgccatggg gacctggatg ctgacgaagg ctcgcgaggc
     1201 tgtgagcagc cacagtgccc tgctcagaag ccccaagctc gtcagtcaag ccggttctcc
     1261 gtttgcactc aggagcacgg gcaggcgagt ggcccctagt tctgggggca gcgggggatc
     1321 cgccaccatg gtttctatcg cactgcaggc tggttatgac ctcctcggag atggtagacc
     1381 tgaaagtctg tggctcggaa ttggtacact gctgatgctg atcggaacct tctacttcct
     1441 ggttagagcg tggggagaga ctgacaagga tgcacgggag tactacgcag tgacaatcct
     1501 ggtcagcggt attgccagtg cagcttatct gtcaatgttc tttggtattg gcctcacaga
     1561 ggtgccagta ggtggagaga tgctgaacat ctactacgca agatacgcac agtggctgtt
     1621 cactacacca ctgttgctgc tgcacctggc actcctggcc aaagttgata gagtgaccat
     1681 cggcactctc gttggcgtcg atgctctgat gattgtcaca ggcctgatag gaacactgtc
     1741 acatactgcc attgccagat actcttggtg gctgttctca actatctgta tgatcgtcgt
     1801 tctgtatgtc ctggccactt ctctgcggtc agccgctaag gagagaggcc cagaggttgc
     1861 ctcaaccttc aacatcctca ccgcactggt tctggtcctg tggactgctt accctatcat
     1921 ctggattatc ggcaccgagg gtgccggagt tgtgggcctg ggaatcgaaa ctttgctgtt
     1981 catggtgctg gacgtcaccg ccaaggtggg cttcggtttt atcctgctca ggtcaagagc
     2041 cattctgcaa gatacagaag cacctgaacc ttcagctgga gctgataccg gtgccgccga
     2101 ccgcccggta gtagcagtga gcaaggcggc cgccaagagc aggatcacca gcgagggcga
     2161 gtacatcccc ctggaccaga tcgacatcaa cgtggtgagc aagggcgagg agctgttcac
     2221 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2281 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatttgcac
     2341 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2401 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2461 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2521 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2581 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2641 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     2701 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2761 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     2821 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2881 cactctcggc atggacgagc tgtacaagtt ctgctacgag aacgaggtgt aagaattcga
     2941 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     3001 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3061 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     3121 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     3181 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     3241 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3301 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     3361 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     3421 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3481 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3541 cccgcatcga taccgtcgac ccgggcggcc gcttcgagca gacatgataa gatacattga
     3601 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3661 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3721 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta
     3781 aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata
     3841 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc
     3901 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     3961 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca
     4021 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4141 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4681 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4741 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4801 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4861 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4921 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4981 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5041 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5101 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5161 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5221 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5281 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5341 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5401 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5461 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5521 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5581 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5641 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5701 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5761 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5821 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5881 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5941 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     6001 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     6061 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     6121 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     6181 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     6241 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     6301 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     6361 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     6421 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     6481 cgatcggtgc gggcctcttc gctattacgc cagctgcgcg ctcgctcgct cactgaggcc
     6541 gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
     6601 gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac
     6661 ccgccatgct acttatctac gtagccatgc tc
//
