LOCUS       sequence-492134-        4757 bp    DNA     circular SYN 06-NOV-2025
DEFINITION  AiE2210m is an enhancer sequence designed to drive AAV-mediated 
            transgene expression in specific population sof brain cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4757)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4757)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP1543 - pAAV-AiE2210m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..483)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(165..186)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     377..396
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   558..698
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   569..698
                     /label=AAV2 ITR
     polyA_signal    816..927
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1114..1134
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1227..1937,1938..1940,1941..1943))
                     /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(1269..1290)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1618..1637
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1877..1898
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1940..1949
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2588..2717
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2779..3367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2859..2878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3538..4329,4330..4398))
                     /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     4161..4180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4399..4503)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4571..4589
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4627..4649)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(4749..4757,1..11)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg
       61 cgttaaattt ttgttaaatc agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc
      121 cttataaatc aaaagaatag cccgagatag agttgagtgt tgttccagtt tggaacaaga
      181 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
      241 atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag
      301 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
      361 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg
      421 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
      481 cgtactatgg ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa
      541 ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
      601 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
      661 cagagaggga gtggccaact ccatcactag gggttcctgc ggccgctcgg tccgcacgtg
      721 agatctcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac
      781 cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg
      841 aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa
      901 cagatggctg gcaactagaa ggcacagtcg aagatctctc gagccacgga attgtcagtg
      961 cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg
     1021 gcaagaacta accaggattt atacaaggag gagaaaatga aagccatacg ggaagcaata
     1081 gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc aacatagtta
     1141 agaataccag tcaatctttc acaaattttg taatccagag gttgattatg atatcgaatt
     1201 cgcggccgcg gcgcgccgtc gacttacttg tacagctcgt ccatgccgag agtgatcccg
     1261 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     1321 agcttggact ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     1381 ggggtgttct gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg
     1441 atcttgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg
     1501 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     1561 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     1621 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     1681 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcacg
     1741 ccgtagccca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     1801 atcagcttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     1861 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     1921 agctcctcgc ccttgctcac catggtggcg accggtagcg ctagcttcga aagatctgga
     1981 tcccagaagc aaatgtaagc aatagatggc tctgccctga cttttatgcc cagcccgagc
     2041 tccgccggcg caacatttaa acccatttaa ccaacagccc taggcttgaa ttcatgagca
     2101 gcaaaggagg ggtggtatgc accctacact gtggctgtgg caggtgccag aaacatctaa
     2161 agggtaaaga taagtgggga ccgtcctaaa ggggaaaggg aaggaggctg gcttcactgg
     2221 tagcatgagc agctttgtga gatataaccc cacccccacg cctccctaca gatttgaaag
     2281 tcagaaggtc tctgctccat cccccgcctc catcagactc caggaagatt caattacatt
     2341 tagagacatg atgtctgcct cccctggatt aatttccgga tcctgccaaa gcttgtcctt
     2401 atctctttct tcttgctgag agtaagaacc ctataaaatt gtctctatca ggaaccaaga
     2461 aatgagccag ggcaaggcaa acataagaag cagtgcctgt gtctctcctc atttagattc
     2521 aggaaggtcc cacgcatgac agctgactgc cgccggcgat tattactctt acaacgcgtg
     2581 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     2641 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc
     2701 gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2761 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2821 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2881 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2941 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3001 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3061 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3121 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3181 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3241 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3301 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3361 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3421 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3481 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3541 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3601 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3661 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3721 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3781 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3841 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3901 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3961 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4021 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4081 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4141 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4201 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4261 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4321 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4381 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4441 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4501 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4561 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     4621 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     4681 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     4741 taactatgcg gcatcag
//
