LOCUS       sequence-502268-        5014 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  AiE2256m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5014)
  TITLE     Allen Institute AAV Enhancer Plasmids
REFERENCE   2  (bases 1 to 5014)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 10, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP1589 - pAAV-AiE2256m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      375..963
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     864..883
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1025..1154
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2050..2059
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2056..2058,2059..2061,2062..2772)
                     /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(2101..2122)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2362..2381)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2709..2730
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2865..2885)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3072..3183
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3301..3441
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3301..3430
                     /label=AAV2 ITR
     rep_origin      3516..3971
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3603..3622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3813..3834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3988..4007)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4107..4129
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4167..4185)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4253..4357
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4358..4426,4427..5014,1..204)
                     /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(4576..4595)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
       61 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
      121 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
      181 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
      241 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
      301 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
      361 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
      421 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      481 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
      541 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      601 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      661 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      721 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      781 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      841 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      901 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      961 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     1021 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac
     1081 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
     1141 cactaggggt tcctgcggcc gcacgcgttg taagagtaat aatcgccggc gttagtagac
     1201 acaaaacatt tattaagttt atagtggaga aatatggccc cagctttctt aaccgaagga
     1261 tggaggtgaa catcaccaac acgggacaaa tcactgcttc ctggtctgga gcactaacaa
     1321 ccatcgtttc tggagcacat ctgcccacag tgcagagcct gggcctctgc aggggaaacc
     1381 actgactcag tgactggacc agcacactgc tgagtgctct cggtgctgcg aggccaccaa
     1441 aggctaacaa ggagaaaagc gctgagacac tgtccttgat taaaggagac tagagaaacg
     1501 tggggctgag ctggatccta gagcacaaag aagagaagag tggaaaaggg tttctttctt
     1561 ctctaacaag gaatgtgcac tgaagtattt caaagtattg aaacaacatg tctaccactt
     1621 atcctcaaat gattgagaga aaaccgtata catgtaggtc aaaccacttg acatttttca
     1681 tgaacaaaga gggagaataa taaggcaaaa atcaagcaaa aaaatgttaa aattcagaaa
     1741 atctgggtta caagcatata gaaaaccttt taagttttgg gtttttttta catattttga
     1801 gaccgtgtag cccaagcttg tctcaaactc agttcgaagc tgaggagagc cctgaacttc
     1861 tgctccacct tcaaatgcaa gctcaggcgc gtgctatccc accctagccg gaccatttgt
     1921 tattttgtac ctttgctata aaatgtttcg ccggcggagc tcgggctggg cataaaagtc
     1981 agggcagagc catctattgc ttacatttgc ttctgggatc cagatctttc gaagctagcg
     2041 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2101 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     2161 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     2221 gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac
     2281 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     2341 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2401 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2461 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     2521 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     2581 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2641 ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag
     2701 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2761 gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca taatcaacct
     2821 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2881 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2941 attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc
     3001 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga
     3061 gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3121 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3181 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3241 gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc
     3301 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3361 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3421 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     3481 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3541 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     3601 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3661 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3721 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3781 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3841 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     3901 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3961 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4021 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4081 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4141 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4201 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4261 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4321 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4381 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4441 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4501 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4561 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4621 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4681 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4741 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4801 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4861 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4921 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4981 gatggaggcg gataaagttg caggaccact tctg
//
