LOCUS       sequence-208092-        6612 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 6612)
  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 6612)
  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-EGFP
FEATURES             Location/Qualifiers
     source          1..6612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   40..169
                     /label=AAV2 ITR
     promoter        242..1530
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1541..1550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2354..2356,2357..3070)
                     /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(2354..2356,2357..3067)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2396..2417)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2657..2676)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3004..3025
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3095..3683
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3148..3168)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3566..3577)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3726..3847
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3737..3756)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3791..3810
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4009..4138
                     /label=AAV2 ITR
     rep_origin      complement(4375..4963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4455..4474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5134..5925,5926..5994))
                     /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     5757..5776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5995..6099)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6125..6580)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6262..6283)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6474..6493
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tgcgcgctcg ctcgctcact
       61 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
      121 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt gtagttaatg
      181 attaacccgc catgctactt atctacgtag ccatgctcta ggaagatcgt aggttaatta
      241 acattatggc cttaggtcac ttcatctcca tggggttctt cttctgattt tctagaaaat
      301 gagatggggg tgcagagagc ttcctcagtg acctgcccag ggtcacatca gaaatgtcag
      361 agctagaact tgaactcaga ttactaatct taaattccat gccttggggg catgcaagta
      421 cgatatacag aaggagtgaa ctcattaggg cagatgacca atgagtttag gaaagaagag
      481 tccagggcag ggtacatcta caccacccgc ccagccctgg gtgagtccag ccacgttcac
      541 ctcattatag ttgcctctct ccagtcctac cttgacggga agcacaagca gaaactggga
      601 caggagcccc aggagaccaa atcttcatgg tccctctggg aggatgggtg gggagagctg
      661 tggcagaggc ctcaggaggg gccctgctgc tcagtggtga cagatagggg tgagaaagca
      721 gacagagtca ttccgtcagc attctgggtc tgtttggtac ttcttctcac gctaaggtgg
      781 cggtgtgata tgcacaatgg ctaaaaagca gggagagctg gaaagaaaca aggacagaga
      841 cagaggccaa gtcaaccaga ccaattccca gaggaagcaa agaaaccatt acagagacta
      901 caagggggaa gggaaggaga gatgaattag cttcccctgt aaaccttaga acccagctgt
      961 tgccagggca acggggcaat acctgtctct tcagaggaga tgaagttgcc agggtaacta
     1021 catcctgtct ttctcaagga ccatcccaga atgtggcacc cactagccgt taccatagca
     1081 actgcctctt tgccccactt aatcccatcc cgtctgttaa aagggcccta tagttggagg
     1141 tgggggaggt aggaagagcg atgatcactt gtggactaag tttgttcgca tccccttctc
     1201 caaccccctc agtacatcac cctgggggaa cagggtccac ttgctcctgg gcccacacag
     1261 tcctgcagta ttgtgtatat aaggccaggg caaagaggag caggttttaa agtgaaaggc
     1321 aggcaggtgt tggggaggca gttaccgggg caacgggaac agggcgtttc ggaggtggtt
     1381 gccatgggga cctggatgct gacgaaggct cgcgaggctg tgagcagcca cagtgccctg
     1441 ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt ttgcactcag gagcacgggc
     1501 aggcgagtgg cccctagttc tgggggcagc gggggatccc gccaccatgg tttctatcgc
     1561 actgcaggct ggttatgacc tcctcggaga tggtagacct gaaagtctgt ggctcggaat
     1621 tggtacactg ctgatgctga tcggaacctt ctacttcctg gttagagcgt ggggagagac
     1681 tgacaaggat gcacgggagt actacgcagt gacaatcctg gtcagcggta ttgccagtgc
     1741 agcttatctg tcaatgttct ttggtattgg cctcacagag gtgccagtag gtggagagat
     1801 gctgaacatc tactacgcaa gatacgcaca gtggctgttc actacaccac tgttgctgct
     1861 gcacctggca ctcctggcca aagttgatag agtgaccatc ggcactctcg ttggcgtcga
     1921 tgctctgatg attgtcacag gcctgatagg aacactgtca catactgcca ttgccagata
     1981 ctcttggtgg ctgttctcaa ctatctgtat gatcgtcgtt ctgtatgtcc tggccacttc
     2041 tctgcggtca gccgctaagg agagaggccc agaggttgcc tcaaccttca acatcctcac
     2101 cgcactggtt ctggtcctgt ggactgctta ccctatcatc tggattatcg gcaccgaggg
     2161 tgccggagtt gtgggcctgg gaatcgaaac tttgctgttc atggtgctgg acgtcaccgc
     2221 caaggtgggc ttcggtttta tcctgctcag gtcaagagcc attctgcaag atacagaagc
     2281 acctgaacct tcagctggag ctgataccgg tgccgccgac cgcccggtag tagcagtgag
     2341 caaggcggcc gccgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2401 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     2461 tgccacctac ggcaagctga ccctgaagtt catttgcacc accggcaagc tgcccgtgcc
     2521 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     2581 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2641 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2701 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2761 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2821 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2881 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2941 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3001 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3061 gtacaagtaa gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     3121 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3181 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3241 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3301 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3361 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3421 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3481 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     3541 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3601 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3661 ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc ttcgagcaga
     3721 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3781 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3841 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     3901 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag
     3961 agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag
     4021 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4081 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4141 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4201 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4261 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4321 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4381 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4441 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4501 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4561 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4621 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4681 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4741 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4801 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4861 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4921 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4981 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5041 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5101 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5161 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5221 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5281 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5341 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5401 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5461 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5521 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5581 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5641 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5701 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5761 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5821 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5881 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5941 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6001 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6061 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6121 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     6181 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     6241 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     6301 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     6361 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     6421 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     6481 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     6541 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg
     6601 cgcaactgtt gg
//
