LOCUS       sequence-490124-        4812 bp    DNA     circular SYN 21-OCT-2025
DEFINITION  AiE2444m 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 4812)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 4812)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP1839 - pAAV-AiE2444m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             567..590
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     regulatory      824..833
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(830..832,833..835,836..1546)
                     /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(875..896)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1136..1155)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1483..1504
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1639..1659)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1846..1957
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2075..2215
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2075..2204
                     /label=AAV2 ITR
     rep_origin      2290..2745
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2377..2396)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2587..2608
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2762..2781)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2881..2903
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2941..2959)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3027..3131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3132..3200,3201..3992)
                     /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(3350..3369)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4163..4751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4652..4671
                     /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 ggatggggaa aaagttttgg
      181 atagtggccc actttaaaac cttttccctc atcatctttt aaccacaaag ggagtgggag
      241 gaataagttg gagaaaattg aatggtaata aggacgggga gggaaaggaa gaaactcgtt
      301 ttctttgtcc ctgtctctgg cacagtaatt tgactaattt caccaggtat tatcattctt
      361 tgcaaaacac ctaaaagcat agtcagaaat attagacaag aacaagactc agttatcttt
      421 tcagacccat catcatattt tctggggcgc ttagtcagat gcattatcac tccgggaaga
      481 tgtgctctgg atgtctttgc aacgtggtgt gtgggtctca gcgcacatac taaatagcta
      541 tcccaggcaa caagaatgag agcccccacc accaccacca ccaccaccac ccagttaaag
      601 gaatcctgtc aggagatgtt ctcaagaatt accagaactg cttttcatgt aaagttggtt
      661 cctacgagga gacggtagag aaaccagatt tctttctaag tggaagcaag caagcctttc
      721 gattttgccg gagctcgggc tgggcataaa agtcagggca gagccatcta ttgcttacat
      781 ttgcttctgg gatccagatc tttcgaagct agcgctaccg gtcgccacca tggtgagcaa
      841 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      901 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      961 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1021 cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     1081 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1141 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1201 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1261 caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     1321 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     1381 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     1441 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1501 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag tcgacggcgc
     1561 gccgcggccg cgaattcgat atcataatca acctctggat tacaaaattt gtgaaagatt
     1621 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     1681 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     1741 gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     1801 tcggctgttg ggcactgaca attccgtggc tcgagagatc ttcgactgtg ccttctagtt
     1861 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     1921 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     1981 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2041 ggcatgagat ctcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     2101 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     2161 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     2221 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     2281 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2341 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     2401 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2461 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     2521 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2581 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     2641 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     2701 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     2761 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     2821 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     2881 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     2941 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3001 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3061 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4801 ttgctcacat gt
//
