LOCUS       sequence-502237-        4790 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  AiE2214m 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 4790)
  TITLE     Allen Institute AAV Enhancer Plasmids
REFERENCE   2  (bases 1 to 4790)
  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: AiP1547 - pAAV-AiE2214m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      335..923
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     824..843
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   985..1114
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1786..1795
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1792..1794,1795..1797,1798..2508)
                     /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(1837..1858)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2098..2117)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2445..2466
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2601..2621)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2808..2919
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3037..3177
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3037..3166
                     /label=AAV2 ITR
     rep_origin      3252..3707
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3339..3358)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3549..3570
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3724..3743)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3843..3865
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3903..3921)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3989..4093
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4094..4162,4163..4790,1..164)
                     /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(4312..4331)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
       61 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
      121 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
      181 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
      241 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
      301 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
      361 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
      421 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
      481 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      541 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      601 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      661 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      721 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      781 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      841 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
      901 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
      961 tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg
     1021 ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     1081 gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgttg taagagtaat
     1141 aatcgccggc gactagggga atatttagta tttaaatggt atgtggtgga tggcataaag
     1201 attatttctg agaatactga gaaatgagaa ccaaagataa atttggaagg gaagtgtttc
     1261 tagcagaaga aggggtttgt gtagagggca tggtacaaaa aagacatact ttatctgaga
     1321 acttcaaagt tcaatgcaac tgaaagaggc ttgaaaaaat atgagagagt tatagatcag
     1381 gatcaaagcc agtgtttgac cactgccagc ttgtggaaca tgagaagatc ttttatttat
     1441 tttagaaaga ggggtatttc ggaagattca taaaatagga gtcagattat tttatgtatt
     1501 ctcttgcttt ggaagagcat gagacaggga tagtagagaa ggggaagtga gggctgttaa
     1561 tatggtcaca aaatccagat gagagatgac agtgagaata gtaaaagtag aaccaagaca
     1621 ttggaaatag aacagagggg ggccttctgc agcagatttc ctatttagag gaaatgggag
     1681 gggacgccgg cggagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac
     1741 atttgcttct gggatccaga tctttcgaag ctagcgctac cggtcgccac catggtgagc
     1801 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1861 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1921 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1981 accctgggct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     2041 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     2101 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     2161 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     2221 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     2281 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     2341 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2401 taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2461 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agtcgacggc
     2521 gcgccgcggc cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga
     2581 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     2641 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     2701 tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2761 gctcggctgt tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag
     2821 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2881 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2941 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3001 caggcatgag atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     3061 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3121 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     3181 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     3241 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3301 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     3361 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3421 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3481 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3541 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     3601 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     3661 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3721 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3781 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3841 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3901 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3961 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4021 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4081 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4141 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4201 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4261 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4321 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4381 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4441 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4501 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4561 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4621 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4681 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4741 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
//
