LOCUS       sequence-323605-        7188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of SomArchon-GFP under the EF1-alpha-1.1 
            promoter, in floxed/reversed (Cre-dependent) manner..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7188)
  AUTHORS   Piatkevich KD, Bensussen S, Tseng HA, Shroff SN, Lopez-Huerta VG, 
            Park D, Jung EE, Shemesh OA, Straub C, Gritton HJ, Romano MF, Costa 
            E, Sabatini BL, Fu Z, Boyden ES, Han X
  TITLE     Population imaging of neural activity in awake behaving mice.
  JOURNAL   Nature. 2019 Oct;574(7778):413-417. doi: 10.1038/s41586-019-1641-1. 
            Epub 2019 Oct 9.
  PUBMED    31597963
REFERENCE   2  (bases 1 to 7188)
  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: 
            "10.1038/s41586-019-1641-1"; journalName: "Nature"; date: "2019-10";
            volume: "574"; issue: "7778"; pages: "413-417"
COMMENT     Sequence Label: AAV-EF1-alpha-1.1-FLEX-rc [SomArchon-GFP]
FEATURES             Location/Qualifiers
     source          1..7188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   47..176
                     /label=AAV2 ITR
     promoter        249..1427
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          480..1418
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1375..1395
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1436..1469
                     /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    1532..1565
                     /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(1819..2529,2530..2532))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(1819..2529,2530..2532))
                     /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(1861..1882)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2210..2229
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2469..2490
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3396..3405
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3416..3449)
                     /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(3520..3553)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3586..4174
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3639..3659)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4057..4068)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4215..4422
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4592..4721
                     /label=AAV2 ITR
     rep_origin      complement(4958..5546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5038..5057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5717..6508,6509..6577))
                     /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     6340..6359
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6578..6682)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6708..7163)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6845..6866)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7057..7076
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgc gcgctcgctc
       61 gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc
      121 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttccttgta
      181 gttaatgatt aacccgccat gctacttatc tacgtagcca tgctctagga agatcgtagg
      241 ttaattaagg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
      301 agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact
      361 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
      421 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
      481 taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc
      541 cttgaattac ttccacctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa
      601 gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg
      661 aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc
      721 tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt
      781 ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt
      841 ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg
      901 gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc
      961 tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt
     1021 cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa
     1081 aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg
     1141 cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc
     1201 acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt
     1261 atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca gcttggcact
     1321 tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc
     1381 ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgagct agcccataac
     1441 ttcgtataaa gtatcctata cgaagttata tcaaaatagg aagaccaatg cttcaccatc
     1501 gacccgaatt gccaagcatc accatcgacc cataacttcg tataatgtat gctatacgaa
     1561 gttatactag aattcttaca cctcgttctc gtagcagaac ttgtagaacc tggtggcctc
     1621 agggaagtcg gtggcgcagc tgatgaagct gtcgatgctg ctcatgctcc tcatgtcgat
     1681 cacgccctcg gtcctggcag gcagaggggc cacagggcta ggcatgctgc tcatctccag
     1741 ctcctcagga ggcttgctct gaggggccat ctccttggtg ttcaggatag gctggctctg
     1801 ggtacctccg ctcccaccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     1861 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga
     1921 ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     1981 ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa
     2041 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
     2101 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     2161 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     2221 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     2281 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
     2341 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc aaatgaactt
     2401 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     2461 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     2521 gcccttgctc accacgttga tgtcgatctg gtccaggggg atgtactcgc cctcgctggt
     2581 gatcctgctc ttggcggccg ccttgctcac tgctactacc gggcggtcgg cggcaccggt
     2641 atcagctcca gctgaaggtt caggtgcttc tgtatcttgc agaatggctc ttgacctgag
     2701 caggataaaa ccgaagccca ccttggcggt gacgtccagc accatgaaca gcaaagtttc
     2761 gattcccagg cccacaactc cggcaccctc ggtgccgata atccagatga tagggtaagc
     2821 agtccacagg accagaacca gtgcggtgag gatgttgaag gttgaggcaa cctctgggcc
     2881 tctctcctta gcggctgacc gcagagaagt ggccaggaca tacagaacga cgatcataca
     2941 gatagttgag aacagccacc aagagtatct ggcaatggca gtatgtgaca gtgttcctat
     3001 caggcctgtg acaatcatca gagcatcgac gccaacgaga gtgccgatgg tcactctatc
     3061 aactttggcc aggagtgcca ggtgcagcag caacagtggt gtagtgaaca gccactgtgc
     3121 gtatcttgcg tagtagatgt tcagcatctc tccacctact ggcacctctg tgaggccaat
     3181 accaaagaac attgacagat aagctgcact ggcaataccg ctgaccagga ttgtcactgc
     3241 gtagtactcc cgtgcatcct tgtcagtctc tccccacgct ctaaccagga agtagaaggt
     3301 tccgatcagc atcagcagtg taccaattcc gagccacaga ctttcaggtc taccatctcc
     3361 gaggaggtca taaccagcct gcagtgcgat agaaaccatg gtggcggatc ctagtataac
     3421 ttcgtatagg atactttata cgaagttatc attgggattc ttcctatttt gatccaagca
     3481 tcaccatcga ccctctagtc cagatctcac catcgaccca taacttcgta tagcatacat
     3541 tatacgaagt tatgtccctc gaagaggttc gcgaagctta tcgataatca acctctggat
     3601 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3661 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3721 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3781 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3841 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3901 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3961 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     4021 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     4081 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     4141 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg
     4201 ctcataaagc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4261 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4321 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4381 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtcc ggactgtaca
     4441 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt
     4501 aaaacctcta caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat
     4561 aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact
     4621 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4681 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcattaa tgaatcggcc
     4741 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4801 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4861 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4921 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4981 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5041 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5101 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5161 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5221 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5281 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5341 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5401 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5461 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5521 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5581 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5641 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5701 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5761 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5821 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5881 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5941 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6001 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6061 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6121 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6181 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6241 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6301 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6361 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6421 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6481 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6541 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6601 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6661 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     6721 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     6781 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     6841 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     6901 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     6961 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     7021 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     7081 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     7141 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaa
//
