LOCUS       sequence-411065-        5196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0390m_3xC2 is an optimized enhancer sequence designed to drive 
            AAV-mediated transgene expression in brain astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5196)
  TITLE     Allen Institute Plasmids Aug 2023
REFERENCE   2  (bases 1 to 5196)
  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: AiP13318 -
            pAAV-AiE0390m_3xC2-minBG-SYFP2-4x2C...
FEATURES             Location/Qualifiers
     source          1..5196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      988..997
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(994..996,997..999,1000..1710)
                     /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(1039..1060)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1300..1319)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1647..1668
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2023..2043)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2230..2341
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2459..2599
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     rep_origin      2674..3129
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2761..2780)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2971..2992
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3146..3165)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3265..3287
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3325..3343)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3411..3515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3516..3584,3585..4376)
                     /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(3734..3753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4547..5135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5036..5055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgaaagg aaagagcttg ctttcaacct caaaagctag
      181 gaggaaaggg ctctgaaatt tgctcagaat tcccaattca ccattagcct gtttcttcct
      241 ttagcctcaa ggcattctcc gctttttgaa aagatgttaa gaaattcagt cacaatagag
      301 agcctagttt tgaacatgtt tcactcggtc cattgaggtc tgggctccag cctttgtgtg
      361 gggtgaattg agctgagcgg ctagctggtt ggaggaaagg aaagagcttg ctttcaacct
      421 caaaagctag gaggaaaggg ctctgaaatt tgctcagaat tcccaattca ccattagcct
      481 gtttcttcct ttagcctcaa ggcattctcc gctttttgaa aagatgttaa gaaattcagt
      541 cacaatagag agcctagttt tgaacatgtt tcactcggtc cattgaggtc tgggctccag
      601 cctttgtgtg gggtgaattg agctgagcgg ctagctggtt ggaggaaagg aaagagcttg
      661 ctttcaacct caaaagctag gaggaaaggg ctctgaaatt tgctcagaat tcccaattca
      721 ccattagcct gtttcttcct ttagcctcaa ggcattctcc gctttttgaa aagatgttaa
      781 gaaattcagt cacaatagag agcctagttt tgaacatgtt tcactcggtc cattgaggtc
      841 tgggctccag cctttgtgtg gggtgaattg agctgagcgg ctagctggtt ggaggagctc
      901 gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca
      961 gatctttcga agctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc
     1021 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1081 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
     1141 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg
     1201 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1261 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1321 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1381 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1441 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     1501 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     1561 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc
     1621 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1681 atcactctcg gcatggacga gctgtacaag taagtcgacg gcgcgccgcg gccttaaaga
     1741 gaccggttca ctgtgacagt aaaagagacc ggttcactgt gagaatgaaa gagaccggtt
     1801 cactgtgatc ggaaaagaga ccggttcact gtgagcggcc ttgaaaccca gcagacaatg
     1861 tagctcagta gaaacccagc agacaatgta gctgaatgga aacccagcag acaatgtagc
     1921 ttcggagaaa cccagcagac aatgtagctg tcgacgaatt cgatatcata atcaacctct
     1981 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2041 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2101 tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg
     2161 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggctcgaga
     2221 gatcttcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2281 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2341 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2401 gggaggattg ggaagacaat agcaggcatg agatctcacg tgcggaccga gcggccgcag
     2461 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     2521 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     2581 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     2641 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     2701 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     2761 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2821 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2881 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2941 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3001 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     3061 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     3121 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     3181 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3241 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3301 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3361 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3421 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3481 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3541 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3601 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3661 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3721 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3781 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3841 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3901 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3961 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4021 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4081 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4141 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4201 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4261 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4321 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4381 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4441 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4501 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4561 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4621 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4681 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4741 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4801 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4861 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4921 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4981 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5041 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5101 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5161 ggttcctggc cttttgctgg ccttttgctc acatgt
//
