LOCUS       sequence-485876-        5675 bp    DNA     circular SYN 12-SEP-2025
DEFINITION  AiE1398m 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 5675)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 5675)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 12, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP15022 -
            pAAV-AiE1398m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5675
                     /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      1028..1037
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1034..1036,1037..1039,1040..1750)
                     /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(1340..1359)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1687..1708
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1760..1816
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1829..1894
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1925..2302
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2389..2409)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2596..2707
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2825..2965
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2825..2954
                     /label=AAV2 ITR
     misc_feature    2972..2999
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    3000..3027
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      3096..3551
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3183..3202)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3393..3414
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3568..3587)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3687..3709
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3747..3765)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3833..3937
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3938..4006,4007..4798)
                     /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(4156..4175)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4969..5557
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5458..5477
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5614..5641
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5642..5669
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtccactc catctcggct gtggctcgcg ggaaaggcag
      181 ccccctacag ttagtctccc cagccatctt ctccggttcc cctgatgctc tcatggcccc
      241 agccatccta gagctggtgc gccatcgctc acctttagca ttcggcctgc tgggggtgag
      301 cttttgggtt agaaggaaca aggcatcaga tggggtcgtg tttttttaca ttccaatcag
      361 ttttcggagg ggaagaacag acatttttgt cagctaattg catttaatta cagagacaat
      421 cgctgtttcc gaacgttgtt gtgtccactc cctcatcctt caatcagttc aagcacatcc
      481 ggccccagct gggagccttt taattgcata atttggattt cacttctgga aagaaacttg
      541 agaaagactc cgtgagggtc tccattaaca caatcagctc cccaggcacc cggggctggc
      601 cctaattgac ccatctcagc taatgcctct ttccaaagaa cgcccgcgtc atttctttgg
      661 cttgtttgtg ctattgtatc ctgttttagt gtcttgatct taaacatctt taaagagggc
      721 actcagtggc ttgtttaact ggaagtggtc cagctggttt ggaaggacag tgcatcaggt
      781 gatgtaagtc caagggtagg aatttctggc ctctccttcc cctgcccatt gcaccgctct
      841 ccccaacagg aagtagctac acatgataag gtgcactgtg gcttctagga cttaggcttt
      901 gcatagtctg tagaccttcc tggggagtgg actggagctc gggctgggca taaaagtcag
      961 ggcagagcca tctattgctt acatttgctt ctgggatcca gatctttcga agctagcgct
     1021 accggtcgcc accatggtct ccaagggcga ggagctgttc accggggtgg tgcccatcct
     1081 ggtcgagctg gacggcgatg tcaacggcca caagttcagc gtgtccggcg agggcgaggg
     1141 cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt
     1201 gccctggccc accctcgtga ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc
     1261 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1321 gcgcaccatc ttcttcaaag acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1381 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1441 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga
     1501 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg
     1561 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1621 gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg
     1681 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1741 gctgtacaag ggctctggtg ctaccaactt ctcactgttg aaacaggcag gggatgtaga
     1801 ggagaatcca gggcctggtg ctagtggaga ctacaaagac catgacggag attataaaga
     1861 tcatgacatc gattacaagg atgacgatga caagtccgga ctcagatctg gaggctccgg
     1921 aggcccagag ccagcgaagt ctgctcccgc cccgaaaaag ggctccaaga aggcggtgac
     1981 taaggcgcag aagaaaggcg gcaagaagcg caagcgcagc cgcaaggaga gctattccat
     2041 ctatgtgtat aaggttctga agcaggtcca ccctgacacc ggcatttcgt ccaaggccat
     2101 gggcatcatg aactcgtttg tgaacgacat tttcgagcgc atcgcaggtg aggcttcccg
     2161 cctggcgcat tacaacaagc gctcgaccat cacctccagg gagatccaga cggccgtgcg
     2221 cctgctgctg cctggggagt tggccaagca cgccgtgtcc gagggtacta aggccatcac
     2281 caagtacacc agcgctaagt aatgaggcgc gccgcggccg cgaattcgat atcataatca
     2341 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2401 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2461 tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc
     2521 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggc
     2581 tcgagagatc ttcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2641 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2701 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2761 gcaaggggga ggattgggaa gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg
     2821 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     2881 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     2941 gagcgagcgc gcagctgcct gcaggggcgc ctatcagcac acaattgccc attatacgcg
     3001 cgtataatgg actattgtgt gctgatatga tgcggtattt tctccttacg catctgtgcg
     3061 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3121 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     3181 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3241 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3301 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3361 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3421 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     3481 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3541 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3601 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3661 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3721 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3781 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3841 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3901 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3961 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4021 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4081 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4141 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4201 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4261 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4321 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4381 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4441 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4501 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4561 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4621 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4681 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4741 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4801 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4861 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4921 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4981 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5041 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5101 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5161 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5221 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5281 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5341 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5401 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5461 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5521 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5581 acggttcctg gccttttgct ggccttttgc tcatatcagc acacaattgc ccattatacg
     5641 cgcgtataat ggactattgt gtgctgataa catgt
//
