LOCUS       sequence-212137-        6346 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  AAV production plasmid encoding for Archon1 fluorescent voltage 
            reporter, Cre-dependent expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6346)
  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 6346)
  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-CAG-FLEX-Archon1-KGC-EGFP-ER2
FEATURES             Location/Qualifiers
     source          1..6346
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     repeat_region   292..421
                     /label=AAV2 ITR
     rep_origin      493..948
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(580..599)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     790..811
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        974..1078
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1079..1147,1148..1939)
                     /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     complement(1297..1316)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2110..2698
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2599..2618
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2935..3064
                     /label=AAV2 ITR
     polyA_signal    3226..3347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3263..3282)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3317..3336
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3390..3978
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3496..3507
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3905..3925
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    4020..4053
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4124..4157
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      4174..4183
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5047..5049,5050..5760)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5047..5049,5050..5760)
                     /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(5089..5110)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5350..5369)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5697..5718
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5809..5842)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(5905..5938)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(5973..5992)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(6054..6076)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
       61 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
      121 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
      181 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg gtactcttcc
      241 tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc
      301 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
      361 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
      421 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
      481 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
      541 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
      601 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
      661 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
      721 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
      781 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
      841 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
      901 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
      961 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1021 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1081 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1141 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     1201 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1261 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1321 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     1381 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     1441 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1501 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1561 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1621 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1681 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1741 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1801 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1861 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1921 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1981 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2041 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2101 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2161 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2221 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2281 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2341 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2401 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2461 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2521 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2581 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2641 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2701 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2761 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2821 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2881 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctgcgc
     2941 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
     3001 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
     3061 tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag
     3121 atccttatcg attttaccac atttgtagag gttttacttg ctttaaaaaa cctcccacat
     3181 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3241 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3301 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgctcg
     3361 aagcggccgc ccgggtcgac ggtatcgatg cggggaggcg gcccaaaggg agatccgact
     3421 cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca ggccgcggga
     3481 aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc cgcgcagaat
     3541 ccaggtggca acacaggcga gcagccaagg aaaggacgat gatttccccg acaacaccac
     3601 ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc aggcggcgat
     3661 gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga gctgacaggt
     3721 ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca ccacgccacg
     3781 ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt tatacaagga
     3841 ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta aagcagcgta
     3901 tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt cacaaatttt
     3961 gtaatccaga ggttgattat cgataagctt gatatcgaat tcgaacctct tcgagggaca
     4021 taacttcgta taatgtatgc tatacgaagt tatgggtcga tggtgagatc tggactagag
     4081 ggtcgatggt gatgcttgga tcaaaatagg aagaatccca atgataactt cgtataaagt
     4141 atcctatacg aagttatact agaattcggt accgccacca tggtttctat cgcactgcag
     4201 gctggttatg acctcctcgg agatggtaga cctgaaagtc tgtggctcgg aattggtaca
     4261 ctgctgatgc tgatcggaac cttctacttc ctggttagag cgtggggaga gactgacaag
     4321 gatgcacggg agtactacgc agtgacaatc ctggtcagcg gtattgccag tgcagcttat
     4381 ctgtcaatgt tctttggtat tggcctcaca gaggtgccag taggtggaga gatgctgaac
     4441 atctactacg caagatacgc acagtggctg ttcactacac cactgttgct gctgcacctg
     4501 gcactcctgg ccaaagttga tagagtgacc atcggcactc tcgttggcgt cgatgctctg
     4561 atgattgtca caggcctgat aggaacactg tcacatactg ccattgccag atactcttgg
     4621 tggctgttct caactatctg tatgatcgtc gttctgtatg tcctggccac ttctctgcgg
     4681 tcagccgcta aggagagagg cccagaggtt gcctcaacct tcaacatcct caccgcactg
     4741 gttctggtcc tgtggactgc ttaccctatc atctggatta tcggcaccga gggtgccgga
     4801 gttgtgggcc tgggaatcga aactttgctg ttcatggtgc tggacgtcac cgccaaggtg
     4861 ggcttcggtt ttatcctgct caggtcaaga gccattctgc aagatacaga agcacctgaa
     4921 ccttcagctg gagctgatac cggtgccgcc gaccgcccgg tagtagcagt gagcaaggcg
     4981 gccgccaaga gcaggatcac cagcgagggc gagtacatcc ccctggacca gatcgacatc
     5041 aacgtggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     5101 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     5161 tacggcaagc tgaccctgaa gttcatttgc accaccggca agctgcccgt gccctggccc
     5221 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     5281 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     5341 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     5401 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     5461 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     5521 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     5581 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     5641 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     5701 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     5761 ttctgctacg agaacgaggt gtaatgtaca agtaagggga tccctagtat aacttcgtat
     5821 agcatacatt atacgaagtt atgggtcgat ggtgatgctt ggcaattcgg gtcgatggtg
     5881 aagcattggt cttcctattt tgatataact tcgtatagga tactttatac gaagttatgg
     5941 ggatccaatt ctttgccaaa atgatgagac agcacaataa ccagcacgtt gcccaggagc
     6001 tgtaggaaaa agaagaaggc atgaacatgg ttagcagagg ctctagagcc gccggtcaca
     6061 cgccagaagc cgaaccccgc cctgccccgt cccccccgaa ggcagccgtc cccccgcgga
     6121 cagccccgag ccgctccccc cgcacaaagg gccctcccgg agcccctcaa ggctttcacg
     6181 cagccacaga aaagaaacaa gccgtcatta aaccaagcgc taattacagc ccggaggaga
     6241 agggccgtcc cgcccgctca cctgtgggag taacgcggtc agtcagagcc ggggcgggcg
     6301 gcgcgaggcg gcggcggagc ggggcacggg gcgaaggcag cgcgca
//
