LOCUS       sequence-245622-        7065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated FLPo-dependent expression of ArchT-GFP under EF1a 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7065)
  AUTHORS   Trouche S, Koren V, Doig NM, Ellender TJ, El-Gaby M, 
            Lopes-Dos-Santos V, Reeve HM, Perestenko PV, Garas FN, Magill PJ, 
            Sharott A, Dupret D
  TITLE     A Hippocampus-Accumbens Tripartite Neuronal Motif Guides Appetitive 
            Memory in Space.
  JOURNAL   Cell. 2019 Mar 7;176(6):1393-1406.e16. doi: 
            10.1016/j.cell.2018.12.037. Epub 2019 Feb 14.
  PUBMED    30773318
REFERENCE   2  (bases 1 to 7065)
  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.cell.2018.12.037"; journalName: "Cell"; date: "2019-03-7-
            7"; volume: "176"; issue: "6"; pages: "1393-1406.e16"
COMMENT     Sequence Label: pAAV-EF1a-FDIO-ArchT-GFP
FEATURES             Location/Qualifiers
     source          1..7065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..126
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        227..1405
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          458..1396
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1353..1373
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1436..1469
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(join(1562..2275,2276..2278))
                     /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             complement(join(1565..2275,2276..2278))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1607..1628)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1956..1975
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2215..2236
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3034..3043
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3050..3083)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3192..3780
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3245..3265)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3663..3674)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3812..4288
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4011..4030)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4328..4468
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4328..4457
                     /label=AAV2 ITR
     rep_origin      4543..4998
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4630..4649)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4840..4861
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5015..5034)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5134..5156
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5194..5212)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5280..5384
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5385..5453,5454..6245)
                     /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(5603..5622)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6416..7004
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6905..6924
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
      121 gttcctgcgg ccgcacgcgt aagctttgca aagatggata aagttttaaa cagagaggaa
      181 tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg
      241 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
      301 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
      361 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
      421 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg
      481 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct
      541 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
      601 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
      661 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
      721 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
      781 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
      841 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
      901 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
      961 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
     1021 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
     1081 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
     1141 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
     1201 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
     1261 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
     1321 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
     1381 ttttcttcca tttcaggtgt cgtgaggtac cggatcctct agagtcgact ccggagaagt
     1441 tcctattctc tagaaagtat aggaacttcg cagaatggta gctggattgt agctgctatt
     1501 agcaatatga aacctcttag aagttcctat tcttcaaaag gtataggaac ttcggcgcgc
     1561 cttacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca
     1621 ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag ggcggactgg gtgctcaggt
     1681 agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt
     1741 cggcgagctg cacgctgccg tcctcgatgt tgtggcggat cttgaagttc accttgatgc
     1801 cgttcttctg cttgtcggcc atgatataga cgttgtggct gttgtagttg tactccagct
     1861 tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca
     1921 ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga
     1981 agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct
     2041 gcttcatgtg gtcggggtag cggctgaagc actgcacgcc gtaggtcagg gtggtcacga
     2101 gggtgggcca gggcacgggc agcttgccgg tggtgcaaat gaacttcagg gtcagcttgc
     2161 cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc
     2221 cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcactg
     2281 ctactgggcc ccggggctcg ggggcctcgg tgtcgcccag gatggccctg ctcctcagca
     2341 ggatgaagcc gaagcccact ttggcggtca cgtccagcac catgaacagc agggtctcga
     2401 ttcccagtcc caccactcca gctccctcgg ttccgatgat ccacaggatg gggtaggcgg
     2461 tccacagcac cagcaccagg gcggtcaggg tgttgaaggt gctagccacc tctggtcccc
     2521 tctccttagc agcagccctc aggctggtag ccaggaagta cagcaccacg atcatgcaga
     2581 tggtgctgaa cagccaccag ctgtacctag ccagtggggt gtggctcagg gcgccgatca
     2641 ggccggtcac gatcatcagg gcgtccactc ccaccagggt tccgatgctc accctgtcca
     2701 ccttagccag cagggccagg tcgagcagaa gcaaaggggt ggtgaacagc cagtcggcgt
     2761 atctggcgta gtagatgtcc agcacctcgc cggccacggt cacctcggtc aggccgatgc
     2821 cgaagaacat gctcaggtag gcggcgctgg cgatgccggg caccaggatt gtgatgctgt
     2881 agtactccct ggcctccttg tcggtcacgc cccagccctt cacgatgaag tagaaggtgc
     2941 cgatcagcat cagcagggtt ccgattccca gccacagggt ctctggcctt ccgtcgccca
     3001 gcaggtcgta tcctgcctgc agagcgatgg ggtccatggt ggcgctagcg aagttcctat
     3061 actttctaga gaataggaac ttcttgcctt aacccagaaa ttatcactgt tattctttag
     3121 aatggtgcaa agagaagttc ctataccttt tgaagaatag gaacttcgaa ttcgatatca
     3181 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3241 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3301 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3361 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3421 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3481 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3541 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     3601 cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3661 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3721 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3781 atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc cctccccagt
     3841 gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt
     3901 aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt
     3961 ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga
     4021 accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca
     4081 agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct
     4141 cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc
     4201 caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc
     4261 gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag
     4321 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4381 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     4441 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     4501 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     4561 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     4621 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4681 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4741 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4801 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4861 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     4921 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4981 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     5041 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     5101 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     5161 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     5221 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     5281 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5341 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5401 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5461 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5521 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5581 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5641 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5701 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5761 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5821 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5881 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5941 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6001 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6061 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6121 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6181 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6241 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6301 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6361 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6421 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6481 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6541 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6601 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6661 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6721 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6781 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6841 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6901 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6961 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7021 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
