LOCUS       sequence-325778-        4679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing SYFP2 AAV Virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4679)
  TITLE     Enhancer AAVs from the Allen Institute
REFERENCE   2  (bases 1 to 4679)
  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"
COMMENT     Sequence Label: AiP11279 - pAAV-eHGT_022m-minBG-SYFP2-WPRE3-BG...
FEATURES             Location/Qualifiers
     source          1..4679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gap             63..84
                     /estimated_length=22
     regulatory      669..678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(675..677,678..680,681..1391)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(720..741)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(981..1000)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1328..1349
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1484..1504)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1684..1795
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2034..2051)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2034..2050)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2043..2065)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2259..2278)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2378..2400
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2438..2456)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2524..2628
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2629..2697,2698..3489)
                     /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(2847..2866)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3660..4248
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4149..4168
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4536..4557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4572..4577,4578..4595,4596..4602)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4610..4626
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4615..4637
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4634..4650
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4634..4650
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaagcttccc ggggggatct gggccactcc ctctctgcgc gctcgctcgc tcactgaggc
       61 cgnnnnnnnn nnnnnnnnnn nnnnycmggg cawwgcccgg gcggcctcag tgagcgagcg
      121 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctggaggg gtggagtcgt
      181 gacctaggac gcgtgtgtcc tccagagtcc gtgggtcctg tgatcccttg ctccattcaa
      241 tgtgctcatc agaagccata tttgaagcag catctaccat ccatggtcca ccttctgtct
      301 ggatccaccc atttaggaaa ccatatcctt gtttgaccag atcactcctg cacacgtgtt
      361 gctttgaatt ctagcacagg attcagcatg acatttccct tggattcctc ttctgagcca
      421 acagagtgag ggctatttcc caaaaaaaca acacattatt aacaaatgaa taagagccaa
      481 gaaaacaata cagccagcat atttagtcca cgcaagcttg tttagaatac aaatcgcaaa
      541 tgtatttaat tcacaaggct cttggggaaa ctttcgagct cgggctgggc ataaaagtca
      601 gggcagagcc atctattgct tacatttgct tctgggatcc agatctttcg aagctagcgc
      661 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      721 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
      781 gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg
      841 tgccctggcc caccctcgtg accaccctgg gctacggcgt gcagtgcttc gcccgctacc
      901 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
      961 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1021 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1081 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg
     1141 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     1201 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1261 tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc aacgagaagc
     1321 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1381 agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc
     1441 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     1501 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     1561 ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct
     1621 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggctcgag
     1681 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1741 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     1801 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     1861 attgggaaga caatagcagg catgactagt ccactccctc tctgcgcgct cgctcgctca
     1921 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     1981 gcgagcgagc gcgcagagag ggacagatcc gggcccgcat gcgtcgacaa ttcactggcc
     2041 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     2101 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     2161 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     2221 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     2281 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2341 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2401 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     2461 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2521 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2581 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2641 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2701 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2761 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2821 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2881 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2941 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3001 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3061 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3121 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3181 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3241 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3301 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3361 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3421 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3481 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3541 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3601 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3661 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3721 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3781 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3841 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3901 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3961 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4021 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4081 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4141 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4201 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4261 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4321 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4381 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     4441 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     4501 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     4561 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     4621 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctctc gagatctag
//
