LOCUS       sequence-502236-        4939 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  AiE2212m 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 4939)
  TITLE     Allen Institute AAV Enhancer Plasmids
REFERENCE   2  (bases 1 to 4939)
  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: AiP1545 - pAAV-AiE2212m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      279..867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     768..787
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   929..1058
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1879..1888
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1885..1887,1888..1890,1891..2601)
                     /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(1930..1951)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2191..2210)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2538..2559
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2694..2714)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2901..3012
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3130..3270
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3130..3259
                     /label=AAV2 ITR
     rep_origin      3345..3800
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3432..3451)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3642..3663
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3817..3836)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3936..3958
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3996..4014)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4082..4186
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4187..4255,4256..4939,1..108)
                     /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(4405..4424)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
       61 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
      121 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
      181 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
      241 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
      301 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
      361 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
      421 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
      481 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
      541 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
      601 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
      661 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
      721 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
      781 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
      841 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
      901 gccttttgct ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact
      961 gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
     1021 cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcacgc gttgtaagag
     1081 taataatcgc cggcgcatga cctgaattag tggaccagct aaatatccca attgctcctt
     1141 agtggaaact ggagcagaaa taaaggtcta tcacacaagt taggaaagca cactggacag
     1201 gcaagcccgg acaggcaagc ccatccagga aacgtgagca cggccttggg gtgtcccctt
     1261 aggatgcggc cacactgggc aggtctgtgg gatggtgagg gctgtgctgg ctgcaggatc
     1321 ataactgcag aggagagagc agcacgtgga tcttccttgt cgtcaggcag aactcttcac
     1381 tgaggtgggc tcagggaggg ctctgtgcct gcgttctggt gagcaatagc cagaagcctt
     1441 gagaggcaga agaggcactg agcaatcgat cagcagctgc tgccccggct aactgaactc
     1501 tctcatccag ctgcaggtgc agagacagtg agacctgacc ctgcctgagc catgccagcc
     1561 aacctcacag agggcagctt tcatgccaac cagactgtgc cgatgctaga ttcttcccca
     1621 gtagcttgca ctgaaattgt gacgttcact gaagcactgg tggctgagga gtggggctcc
     1681 ttctactcct cctttaaggt aagtgccctg gcttcagagg ctggaacagg tgggacagga
     1741 gagagctggg agagtcttag ctgctgctag ggaggctcgc cggcggagct cgggctgggc
     1801 ataaaagtca gggcagagcc atctattgct tacatttgct tctgggatcc agatctttcg
     1861 aagctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     1921 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1981 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
     2041 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt gcagtgcttc
     2101 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2161 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2221 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2281 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     2341 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     2401 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     2461 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc
     2521 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2581 ggcatggacg agctgtacaa gtaagtcgac ggcgcgccgc ggccgcgaat tcgatatcat
     2641 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2701 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2761 atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc
     2821 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     2881 gtggctcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2941 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3001 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3061 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg
     3121 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     3181 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     3241 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     3301 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     3361 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3421 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3481 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3541 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3601 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3661 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     3721 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3781 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     3841 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     3901 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     3961 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     4021 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     4081 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4141 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4201 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4261 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4321 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4381 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4441 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4501 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4561 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4621 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4681 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4741 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4801 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4861 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4921 agcactgggg ccagatggt
//
