LOCUS       sequence-226683-        6283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of Archon1-KGC-GFP-ER2 under the Syn 
            promoter, in floxed/reversed (Cre-dependent) manner. Using bGHpA 
            signal..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6283)
  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 6283)
  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-Syn-FLEX-rc [Archon1-KGC-GFP-ER2]
FEATURES             Location/Qualifiers
     source          1..6283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   83..212
                     /label=AAV2 ITR
     promoter        312..759
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    772..805
                     /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)."
     protein_bind    868..901
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(939..1649,1650..1652))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(939..1649,1650..1652))
                     /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(981..1002)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1330..1349
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1589..1610
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2516..2525
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2547..2580)
                     /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)."
     protein_bind    complement(2651..2684)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2717..3305
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2770..2790)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3188..3199)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3346..3553
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3723..3852
                     /label=AAV2 ITR
     rep_origin      3924..4379
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4011..4030)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4221..4242
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4405..4509
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4510..4578,4579..5370)
                     /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(4728..4747)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5541..6129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6030..6049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
       61 gttggccgat tcattaatgc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
      121 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
      181 gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta
      241 cttatctacg tagccatgct ctaggaagat cgtaggttaa ttaatctaga ctgcagaggg
      301 ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac
      361 ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat
      421 cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct
      481 tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc
      541 actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt
      601 ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg
      661 gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg
      721 gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cgagaattcc cataacttcg
      781 tataaagtat cctatacgaa gttatatcaa aataggaaga ccaatgcttc accatcgacc
      841 cgaattgcca agcatcacca tcgacccata acttcgtata atgtatgcta tacgaagtta
      901 tactagctag ctcattacac ctcgttctcg tagcagaact tgtacagctc gtccatgccg
      961 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     1021 gggtctttgc tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg
     1081 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg
     1141 atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata
     1201 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
     1261 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
     1321 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag
     1381 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg
     1441 aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg
     1501 ccggtggtgc aaatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
     1561 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
     1621 accccggtga acagctcctc gcccttgctc accacgttga tgtcgatctg gtccaggggg
     1681 atgtactcgc cctcgctggt gatcctgctc ttggcggccg ccttgctcac tgctactacc
     1741 gggcggtcgg cggcaccggt atcagctcca gctgaaggtt caggtgcttc tgtatcttgc
     1801 agaatggctc ttgacctgag caggataaaa ccgaagccca ccttggcggt gacgtccagc
     1861 accatgaaca gcaaagtttc gattcccagg cccacaactc cggcaccctc ggtgccgata
     1921 atccagatga tagggtaagc agtccacagg accagaacca gtgcggtgag gatgttgaag
     1981 gttgaggcaa cctctgggcc tctctcctta gcggctgacc gcagagaagt ggccaggaca
     2041 tacagaacga cgatcataca gatagttgag aacagccacc aagagtatct ggcaatggca
     2101 gtatgtgaca gtgttcctat caggcctgtg acaatcatca gagcatcgac gccaacgaga
     2161 gtgccgatgg tcactctatc aactttggcc aggagtgcca ggtgcagcag caacagtggt
     2221 gtagtgaaca gccactgtgc gtatcttgcg tagtagatgt tcagcatctc tccacctact
     2281 ggcacctctg tgaggccaat accaaagaac attgacagat aagctgcact ggcaataccg
     2341 ctgaccagga ttgtcactgc gtagtactcc cgtgcatcct tgtcagtctc tccccacgct
     2401 ctaaccagga agtagaaggt tccgatcagc atcagcagtg taccaattcc gagccacaga
     2461 ctttcaggtc taccatctcc gaggaggtca taaccagcct gcagtgcgat agaaaccatg
     2521 gtggcggcgc ggcctaggta cctagtataa cttcgtatag gatactttat acgaagttat
     2581 cattgggatt cttcctattt tgatccaagc atcaccatcg accctctagt ccagatctca
     2641 ccatcgaccc ataacttcgt atagcataca ttatacgaag ttatgtccct cgaagaggtt
     2701 cgcgaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2761 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2821 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2881 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2941 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     3001 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3061 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     3121 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     3181 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3241 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3301 cccgcatcga taccgtcgac ccgggcggcc gctcataaag ctcgactgtg ccttctagtt
     3361 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3421 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3481 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3541 ggcatgctgg ggatgcggtc cggactgtac aattgcattc attttatgtt tcaggttcag
     3601 ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     3661 aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta
     3721 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3781 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3841 gcgagcgcgc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3901 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3961 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4021 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4081 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4141 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4201 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4261 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4321 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     4381 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4441 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4501 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4561 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4621 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4681 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4741 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4801 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4861 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4921 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4981 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5041 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5101 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5161 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5221 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5281 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5341 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5401 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5461 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5521 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5581 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5641 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5701 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5761 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5821 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5881 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5941 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6001 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6061 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6121 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6181 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6241 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gca
//
