LOCUS       sequence-429013-        7581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ArchT-GFP dependent on the combined activity of CRE- and 
            FLP-recombinases.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7581)
  AUTHORS   Rangel Guerrero DK, Balueva K, Barayeu U, Baracskay P, Gridchyn I, 
            Nardin M, Roth CN, Wulff P, Csicsvari J
  TITLE     Hippocampal cholecystokinin-expressing interneurons regulate 
            temporal coding and contextual learning.
  JOURNAL   Neuron. 2024 Apr 10:S0896-6273(24)00197-1. doi: 
            10.1016/j.neuron.2024.03.019.
  PUBMED    38636524
REFERENCE   2  (bases 1 to 7581)
  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: "Neuron. 
            2024 Apr 10:S0896-6273(24)00197-1. doi: 
            10.1016/j.neuron.2024.03.019."
COMMENT     Sequence Label: pAAV-C+F-ArchT-GFP
FEATURES             Location/Qualifiers
     source          1..7581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   7..136
                     /label=AAV2 ITR
     promoter        442..719
                     /label=chicken beta-actin promoter
     primer_bind     998..1020
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1082..1101
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1144..1191
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     intron          1281..1346
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1476..1496
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1921..2055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1958..1977)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2012..2031
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    2121..2168
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    2177..2210
                     /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    2273..2306
                     /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(2333..3046,3047..3049))
                     /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(2336..3046,3047..3049))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2378..2399)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2727..2746
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2986..3007
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3805..3814
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3840..3873)
                     /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(3944..3977)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4022..4039)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4028..4252
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4462..4791)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4476..4611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4776..4796
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5037..5625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5117..5136)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5796..6587,6588..6656))
                     /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     6419..6438
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6657..6761)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6829..6847
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6885..6907)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7007..7026
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7240..7259
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        7549..7567
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7549..7566
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ttctagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      181 tgctctaggt accattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      241 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      301 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      361 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      421 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      481 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      541 tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
      601 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      661 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      721 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      781 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      841 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      901 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ctgtccgcgg
      961 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1021 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1081 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgga tcggcgccaa
     1141 ttggaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt ccgatcgctc
     1201 gagggatctt tgtgaaggaa ccttacttct gtggtgtgac ataattggac aaactaccta
     1261 cagagattta aagctctaag gtaaatataa aatttttaag tgtataatgt gttaaactac
     1321 tgattctaat tgtttgtgta ttttagattc caacctatgg aactgatgaa tgggagcagt
     1381 ggtggaatgc ctttaatgag gaaaacctgt tttgctcaga agaaatgcca tctagtgatg
     1441 atgaggctac tgctgactct caacattcta ctcctccaaa aaagaagaga aaggtagaag
     1501 accccaagga ctttccttca gaattgctaa gttttttgag tcatgctgtg tttagtaata
     1561 gaactcttgc ttgctttgct atttacacca caaaggaaaa agctgcactg ctatacaaga
     1621 aaattatgga aaaatattct gtaaccttta taagtaggca taacagttat aatcataaca
     1681 tactgttttt tcttactcca cacaggcata gagtgtctgc tattaataac tatgctcaaa
     1741 aattgtgtac ctttagcttt ttaatttgta aaggggttaa taaggaatat ttgatgtata
     1801 gtgccttgac tagagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     1861 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     1921 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     1981 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2041 tatcatgtct ggatctgaca tggtaagtaa gcttgggctg caggtcgagg gacctaacgc
     2101 gtctcgagtc gcgattataa gaagttccta ttccgaagtt cctattctct agaaagtata
     2161 ggaacttcgg atccccataa cttcgtataa agtatcctat acgaagttat atcaaaatag
     2221 gaagaccaat gcttcaccat cgacccgaat tgccaagcat caccatcgac ccataacttc
     2281 gtataatgta tgctatacga agttatgaat tcctgcagcc cgggggatcc ccttacttgt
     2341 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     2401 gatcgcgctt ctcgttgggg tctttgctca gggcggactg ggtgctcagg tagtggttgt
     2461 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     2521 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
     2581 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     2641 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     2701 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     2761 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     2821 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc
     2881 agggcacggg cagcttgccg gtggtgcaaa tgaacttcag ggtcagcttg ccgtaggtgg
     2941 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     3001 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcact gctactaccg
     3061 gtcggggctc gggggcctcg gtgtcgccca ggatggccct gctcctcagc aggatgaagc
     3121 cgaagcccac tttggcggtc acgtccagca ccatgaacag cagggtctcg attcccagtc
     3181 ccaccactcc agctccctcg gttccgatga tccacaggat ggggtaggcg gtccacagca
     3241 ccagcaccag ggcggtcagg gtgttgaagg tgctagccac ctctggtccc ctctccttag
     3301 cagcagccct caggctggta gccaggaagt acagcaccac gatcatgcag atggtgctga
     3361 acagccacca gctgtaccta gccagtgggg tgtggctcag ggcgccgatc aggccggtca
     3421 cgatcatcag ggcgtccact cccaccaggg ttccgatgct caccctgtcc accttagcca
     3481 gcagggccag gtcgagcaga agcaaagggg tggtgaacag ccagtcggcg tatctggcgt
     3541 agtagatgtc cagcacctcg ccggccacgg tcacctcggt caggccgatg ccgaagaaca
     3601 tgctcaggta ggcggcgctg gcgatgccgg gcaccaggat tgtgatgctg tagtactccc
     3661 tggcctcctt gtcggtcacg ccccagccct tcacgatgaa gtagaaggtg ccgatcagca
     3721 tcagcagggt tccgattccc agccacaggg tctctggcct tccgtcgccc agcaggtcgt
     3781 atcctgcctg cagagcgatg gggtccatgg tggcgggccg cctcgagaga tctgatatca
     3841 taacttcgta taggatactt tatacgaagt tatcattggg attcttccta ttttgatcca
     3901 agcatcacca tcgaccctct agtccagatc tcaccatcga cccataactt cgtatagcat
     3961 acattatacg aagttatgtc cctcgaagag gttcaagctt gggggtcgac tcgctgatca
     4021 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4081 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4141 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4201 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4261 gcggaaagaa ccagctgggg ctcgactaga gcatggctac gtagataagt agcatggcgg
     4321 gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     4381 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga
     4441 gcgagcgagc gcgcagagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     4501 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     4561 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     4621 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     4681 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     4741 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     4801 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4861 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4921 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4981 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5041 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5101 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5161 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5221 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5281 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5341 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5401 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5461 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5521 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5581 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5641 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5701 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5761 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5821 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5881 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5941 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6001 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6061 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6121 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6181 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6241 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6301 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6361 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6421 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6481 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6541 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6601 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6661 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6721 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6781 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     6841 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     6901 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     6961 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     7021 attgtactga gagtgcacca ttcgacgctc tcccttatgc gactcctgca ttaggaagca
     7081 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     7141 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     7201 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     7261 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     7321 ctggctagcg atgaccctgc tgattggttc gctgaccatt tccgggtgcg ggacggcgtt
     7381 accagaaact cagaaggttc gtccaaccaa accgactctg acggcagttt acgagagaga
     7441 tgatagggtc tgcttcagta agccagatgc tacacaatta ggcttgtaca tattgtcgtt
     7501 agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca
     7561 ctatagaata cacggaatta a
//
