LOCUS       sequence-490257-        4746 bp    DNA     circular SYN 22-OCT-2025
DEFINITION  AiE2321m 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 4746)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4746)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP1682 - pAAV-AiE2321m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4746
                     /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      758..767
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(764..766,767..769,770..1480)
                     /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(809..830)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1070..1089)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1417..1438
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1573..1593)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1780..1891
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2009..2149
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2009..2138
                     /label=AAV2 ITR
     rep_origin      2224..2679
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2311..2330)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2521..2542
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2696..2715)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2815..2837
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2875..2893)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2961..3065
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3066..3134,3135..3926)
                     /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(3284..3303)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4097..4685
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4586..4605
                     /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 gcgttgtaag agtaataatc ggacagggat ctcccatcct
      181 agacatcact ggtgcgggga gggaggaggg caatgatctc acagtgggct tctcacacgg
      241 aaagcagaag gaaccaggag ttagagcatg ctcctgtgag catggggact cacacatagg
      301 caggtctgct gttcacacag gtccttgtac acatgtagcc caaactccag ctgactcaag
      361 ggctggactt aaggatttca tttcatttcc taaatactga acaaacttcc tgttatagct
      421 gagtgaattt gagatattcc tgtttcttga cattaatttc ctgttgtacc ttttaggggt
      481 gtgcagagat tattataagg gaggctgagc cactgctggt gacagatcga gctctacagc
      541 atccggttta tatcttattc ttatttatat ctgtcttctc ttgtcattta cagtgactga
      601 ggggggctgt gctatcatct cagcaagcca agaagagggc ctgtgttagt ttttctggta
      661 aacggagctc gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt
      721 ctgggatcca gatctttcga agctagcgct accggtcgcc accatggtga gcaagggcga
      781 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      841 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      901 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg
      961 ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1021 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1081 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1141 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1201 caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     1261 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa
     1321 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     1381 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1441 cgccgccggg atcactctcg gcatggacga gctgtacaag taagtcgacg gcgcgccgcg
     1501 gccgcgaatt cgatatcata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1561 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     1621 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttagt
     1681 tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     1741 gttgggcact gacaattccg tggctcgaga gatcttcgac tgtgccttct agttgccagc
     1801 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     1861 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     1921 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     1981 agatctcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     2041 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     2101 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     2161 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     2221 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2281 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2341 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2401 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     2461 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2521 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     2581 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     2641 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     2701 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     2761 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     2821 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     2881 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     2941 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3001 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3061 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3121 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3181 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3241 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3301 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3361 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3421 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3481 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3541 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3601 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3661 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3721 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3781 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3841 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3901 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3961 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4021 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4081 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4141 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4201 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4261 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4321 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4381 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4441 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4501 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4561 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4621 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4681 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4741 acatgt
//
