LOCUS       sequence-204559-        6654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV production plasmid encoding for Archon1 fluorescent voltage 
            reporter, Cre-dependent expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6654)
  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 6654)
  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..6654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1141..1174
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1245..1278
                     /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      1295..1304
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2168..2170,2171..2881)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2168..2170,2171..2881)
                     /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(2210..2231)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2471..2490)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2818..2839
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2930..2963)
                     /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(3026..3059)
                     /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(3094..3113)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3175..3197)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3477..3754)
                     /label=chicken beta-actin promoter
     repeat_region   4067..4196
                     /label=AAV2 ITR
     rep_origin      4268..4723
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4355..4374)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4565..4586
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4749..4853
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4854..4922,4923..5714)
                     /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(5072..5091)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5885..6473
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6374..6393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct tgatatcgaa ttcgaacctc ttcgagggac
     1141 ataacttcgt ataatgtatg ctatacgaag ttatgggtcg atggtgagat ctggactaga
     1201 gggtcgatgg tgatgcttgg atcaaaatag gaagaatccc aatgataact tcgtataaag
     1261 tatcctatac gaagttatac tagaattcgg taccgccacc atggtttcta tcgcactgca
     1321 ggctggttat gacctcctcg gagatggtag acctgaaagt ctgtggctcg gaattggtac
     1381 actgctgatg ctgatcggaa ccttctactt cctggttaga gcgtggggag agactgacaa
     1441 ggatgcacgg gagtactacg cagtgacaat cctggtcagc ggtattgcca gtgcagctta
     1501 tctgtcaatg ttctttggta ttggcctcac agaggtgcca gtaggtggag agatgctgaa
     1561 catctactac gcaagatacg cacagtggct gttcactaca ccactgttgc tgctgcacct
     1621 ggcactcctg gccaaagttg atagagtgac catcggcact ctcgttggcg tcgatgctct
     1681 gatgattgtc acaggcctga taggaacact gtcacatact gccattgcca gatactcttg
     1741 gtggctgttc tcaactatct gtatgatcgt cgttctgtat gtcctggcca cttctctgcg
     1801 gtcagccgct aaggagagag gcccagaggt tgcctcaacc ttcaacatcc tcaccgcact
     1861 ggttctggtc ctgtggactg cttaccctat catctggatt atcggcaccg agggtgccgg
     1921 agttgtgggc ctgggaatcg aaactttgct gttcatggtg ctggacgtca ccgccaaggt
     1981 gggcttcggt tttatcctgc tcaggtcaag agccattctg caagatacag aagcacctga
     2041 accttcagct ggagctgata ccggtgccgc cgaccgcccg gtagtagcag tgagcaaggc
     2101 ggccgccaag agcaggatca ccagcgaggg cgagtacatc cccctggacc agatcgacat
     2161 caacgtggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2221 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2281 ctacggcaag ctgaccctga agttcatttg caccaccggc aagctgcccg tgccctggcc
     2341 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2401 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2461 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2521 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2581 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2641 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2701 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2761 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     2821 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2881 gttctgctac gagaacgagg tgtaatgtac aagtaagggg atccctagta taacttcgta
     2941 tagcatacat tatacgaagt tatgggtcga tggtgatgct tggcaattcg ggtcgatggt
     3001 gaagcattgg tcttcctatt ttgatataac ttcgtatagg atactttata cgaagttatg
     3061 gggatccaat tctttgccaa aatgatgaga cagcacaata accagcacgt tgcccaggag
     3121 ctgtaggaaa aagaagaagg catgaacatg gttagcagag gctctagagc cgccggtcac
     3181 acgccagaag ccgaaccccg ccctgccccg tcccccccga aggcagccgt ccccccgcgg
     3241 acagccccga gccgctcccc ccgcacaaag ggccctcccg gagcccctca aggctttcac
     3301 gcagccacag aaaagaaaca agccgtcatt aaaccaagcg ctaattacag cccggaggag
     3361 aagggccgtc ccgcccgctc acctgtggga gtaacgcggt cagtcagagc cggggcgggc
     3421 ggcgcgaggc ggcggcggag cggggcacgg ggcgaaggca gcgtcgcagc gactcccgcc
     3481 cgccgcgcgc ttcgcttttt atagggccgc cgccgccgcc gcctcgccat aaaaggaaac
     3541 tttcggagcg cgccgctctg attggctgcc gccgcacctc tccgcctcgc cccgccccgc
     3601 ccctcgcccc gccccgcccc gcctggcgcg cgcccccccc ccccccccgc ccccatcgct
     3661 gcacaaaata attaaaaaat aaataaatac aaaattgggg gtggggaggg gggggagatg
     3721 gggagagtga agcagaacgt ggggctcacc tcgaccatgg taatagcgat gactaatacg
     3781 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
     3841 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
     3901 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     3961 attggcgtta ctatgggaac atacgtcatt attgacgtca atggtacgat cttcctagag
     4021 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg
     4081 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4141 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgg
     4201 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4261 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4321 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4381 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4441 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4501 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4561 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4621 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4681 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     4741 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4801 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4861 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4921 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4981 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5041 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5101 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5161 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5221 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5281 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5341 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5401 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5461 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5521 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5581 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5641 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5701 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5761 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5821 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5881 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5941 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6001 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6061 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6121 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6181 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6241 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6301 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6361 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6421 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6481 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6541 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6601 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaag
//
