LOCUS       sequence-248519-        7598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV Flex reverse plasmid with the coding sequence of eGFP fused to 
            the N-term of the FMRFamide gated Na channel.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7598)
  AUTHORS   Schanuel SM, Bell KA, Henderson SC, McQuiston AR
  TITLE     Heterologous expression of the invertebrate FMRFamide-gated sodium 
            channel as a mechanism to selectively activate mammalian neurons.
  JOURNAL   Neuroscience. 2008 Aug 13;155(2):374-86. doi: 
            10.1016/j.neuroscience.2008.05.052. Epub 2008 Jun 10.
  PUBMED    18598740
REFERENCE   2  (bases 1 to 7598)
  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"; doi: 
            "10.1016/j.neuroscience.2008.05.052"; journalName: "Neuroscience"; 
            date: "2008-08-13- 13"; volume: "155"; issue: "2"; pages: "374-86"
COMMENT     Sequence Label: AAV-Flex-rev-GFPFANACfullWT
FEATURES             Location/Qualifiers
     source          1..7598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        441..718
                     /label=chicken beta-actin promoter
     primer_bind     997..1019
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1081..1100
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1135..1168
                     /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    1231..1264
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3175..3840)
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     CDS             complement(3175..3840)
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     protein_bind    complement(3863..3896)
                     /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(3967..4000)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4042..4630
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4095..4115)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4513..4524)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4673..4794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4684..4703)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4738..4757
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4956..5085
                     /label=AAV2 ITR
     rep_origin      complement(5322..5910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5402..5421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6081..6872,6873..6941))
                     /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     6704..6723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6942..7046)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7072..7527)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7209..7230)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7421..7440
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg ggcgcgcgcc gggggggggg gggggggggg gggggggggg
      601 sggggsgrgg cggrgaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      661 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      721 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      781 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      841 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      901 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg
      961 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1021 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1081 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattggat ccccataact
     1141 tcgtataaag tatcctatac gaagttatat caaaatagga agaccaatgc ttcaccatcg
     1201 acccgaattg ccaagcatca ccatcgaccc ataacttcgt ataatgtatg ctatacgaag
     1261 ttatactagg atcccctcga gtcatacagg ggactcgacc ccccgtctaa agtcaaacac
     1321 cgaggctcca gctgagtcgg agtagtgact gtcgtggtcc atgtagccgt tgtagagctg
     1381 atgctgactg gtactttggc tgtggttgtt gctgccgttg ttgttgttga cggtggtcgg
     1441 gcctcggggc agccccttct cggagttgaa gaccaggcca gtcagtctga tgaccagctc
     1501 gatgagctcc atgatggtca gcacggagat gcccatccac agccccagag tgccgccgat
     1561 gtcggcgaac aggtcggcca ggccgtaggc cggcagctgg cggtactcca ccacactcag
     1621 gtcctccaag taaatattca acctcagcat gttctgacgg atgaggtccg aggcctcttt
     1681 agccatctct ttctccgata gagagtagct cttggacaga atgtcgtcca tgtggctgtc
     1741 gttcctggcc aggtgcttct ggctgggatg ggccagtttc tccaggtact cgtaggccgt
     1801 cttcatgaag tgattctggc cagcttgtct ttcttgtttg aaaaacctct ctactgcact
     1861 gagctggtag aactccagcg gccagtagga cagcgacaca gatttcaggt aggacgtctc
     1921 actgcacggc tggaagcagc cacagctcag ctcgtacgtc ctgtcgttgt tgaggttctt
     1981 ctgctcccgc tcctcgcagg ccaggtatgg cgtggggatg aaggctcggt cctcttccga
     2041 ctggttgtgg tcttcattgg aatgattcct gttgatttct tgccagtcct taatgacgcc
     2101 acagaaggtt gcgttgtagc tgggcacctc tggtagggcc gaggatttgc agccacagcg
     2161 ctggataatg agtcgctgtt tgcacagctg caggcatgcg aagaaagtgt acttgtactg
     2221 cttgatgcca ttcagcatgt cgtttgtaca gttgccgtaa ggatatggta ggcgcgtgtg
     2281 aagaatagcc ttcaagccta cggacgaaga atatccagga ggaatatcaa tcccatgatc
     2341 gacaggactg ggcatggagc cgggggcgtg caccacaaca cgaaccccgg cgctgtgcag
     2401 gatgttgttg tcgaagttat acacaccata agtgcccggc agtgggtcgt ctttctccac
     2461 agagaagatc agggacagcc cattctctgg tccagtagcg tgcatctgta gcctctggcc
     2521 gctgttgaag gtgaaacagt tgaagtagtt cccatcaaag aatgtgctga agttggacac
     2581 gtggcagagt tcccggttaa agcggcagtg catcaacata tcctccaggt tgtgactcag
     2641 tttcttggcg tcctgaccca gattctcgta gaacgcccgg atactgttca taaaggagtc
     2701 ttgctcaaac cggaacttct gaatgaaccg taaccacgtg attaagtttt ggctctcatt
     2761 gttgaagtac atccggcgaa tagtgcggag ggagattggt tcgatgttgc agatggacac
     2821 agaagggtac tgtaccggca tgctgtcctt gatctctgac agctccacca cctgaaactg
     2881 caggtacttc ctgaccagca gcgacagctg cagcgtggca gccgtgaagc cggcgatcac
     2941 cagcaatgcc cagatgacct ttctcttggt gtctcgcgag gtgacgatct tggccagccc
     3001 gtgggcgttg ctctcggagc ccagttcagc gatgatgtcg atggcgctgc ggttgtcgga
     3061 gtagttgcag tgatgtgggt gatatcgatt cctcatcatc gcgtgctgat gatgttcagg
     3121 aaacactccc ggcttggtgg cagcgcttgt gtatttcatt cgagctcggg atctggcgaa
     3181 ggcgatgggg gtcttgaagg cgtgctggta ctccacgatg cccagctcgg tgttgctgtg
     3241 cagctcctcc acgcggcgga aggcgaacat ggggcccccg ttctgcagga tgctggggtg
     3301 gatggcgctc ttgaagtgca tgtggctgtc caccacgaag ctgtagtagc cgccgtcgcg
     3361 caggctgaag gtgcgggcga agctgcccac cagcacgtta tcgcccatgg ggtgcaggtg
     3421 ctccacggtg gcgttgctgc ggatgatctt gtcggtgaag atcacgctgt cctcggggaa
     3481 gccggtgccc accaccttga agtcgccgat cacgcggccg gcctcgtagc ggtagctgaa
     3541 gctcacgtgc agcacgccgc cgtcctcgta cttctcgatg cgggtgttgg tgtagccgcc
     3601 gttgttgatg gcgtgcagga aggggttctc gtagccgctg gggtaggtgc cgaagtggta
     3661 gaagccgtag cccatcacgt ggctcagcag gtaggggctg aaggtcaggg cgcctttggt
     3721 gctcttcatc ttgttggtca tgcggccctg ctcgggggtg ccctctccgc cgcccaccag
     3781 ctcgaactcc acgccgttca gggtgccggt gatgcggcac tcgatcttca tggcgggcat
     3841 ggtttaggta ccgaattcta gtataacttc gtataggata ctttatacga agttatcatt
     3901 gggattcttc ctattttgat ccaagcatca ccatcgaccc tctagtccag atctcaccat
     3961 cgacccataa cttcgtatag catacattat acgaagttat gtccctcgaa gaggttcgaa
     4021 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     4081 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     4141 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4201 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4261 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     4321 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     4381 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     4441 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     4501 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     4561 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     4621 gcctccccgc atcgataccg tcgacccggg cggccgcttc gagcagacat gataagatac
     4681 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     4741 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     4801 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     4861 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt
     4921 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     4981 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     5041 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc
     5101 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5161 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5221 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5281 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5341 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5401 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5461 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5521 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5581 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5641 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5701 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5761 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5821 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5881 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5941 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6001 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6061 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6121 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6181 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6241 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6301 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6361 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6421 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6481 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6541 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6601 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6661 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6721 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6781 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6841 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6901 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6961 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7021 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     7081 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     7141 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     7201 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     7261 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     7321 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     7381 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     7441 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     7501 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     7561 aagggcgatc ggtgcgggcc tcttcgctat tacgccag
//
