LOCUS       sequence-489678-        5270 bp    DNA     circular SYN 17-OCT-2025
DEFINITION  AiE1045m 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 5270)
  TITLE     Basal Ganglia/CERP Enhancer AAVs
REFERENCE   2  (bases 1 to 5270)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP14974 -
            pAAV-AiE1045m-SYFP2-P2A-3XFLAG-H2B-...
FEATURES             Location/Qualifiers
     source          1..5270
                     /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      623..632
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(629..631,632..634,635..1345)
                     /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(935..954)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1282..1303
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1355..1411
                     /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             1424..1489
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1520..1897
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(1984..2004)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2191..2302
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2420..2560
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2420..2549
                     /label=AAV2 ITR
     misc_feature    2567..2594
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2595..2622
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2691..3146
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2778..2797)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2988..3009
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3163..3182)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3282..3304
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3342..3360)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3428..3532
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3533..3601,3602..4393)
                     /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(3751..3770)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4564..5152
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5053..5072
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5209..5236
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5237..5264
                     /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 gcgttcaatg cctttgttcc ggagtctgtt agagaagaaa
      181 aagttacagt agcataacaa agcaggaaaa gggaatggaa aaaaaaaata ataactggct
      241 tccagtgtct ggagcagctg cacacacacc cttagcacag gtttggcttg gtaaattaat
      301 tcatctgtaa atagtgagca gtctccagat agaacttcag ggctgaggtg caggttcctt
      361 aaggatttac atgtaaaggc aaaggctttc agccctaggt gttacaagtg agtggtgccc
      421 ttcatgccct gccctgagtc tgcccatcca atcatatccc gatatgcaaa tgacccttag
      481 tgcatgcaga taaaaaggag caagggctct ttcagaacca atttgctgag cctgtcacct
      541 gtgttccagg agccaaacca gcaatgtctt cgcctgcaca acctgcaggg atccagatct
      601 ttcgaagcta gcgctaccgg tcgccaccat ggtctccaag ggcgaggagc tgttcaccgg
      661 ggtggtgccc atcctggtcg agctggacgg cgatgtcaac ggccacaagt tcagcgtgtc
      721 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
      781 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg
      841 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
      901 aggctacgtc caggagcgca ccatcttctt caaagacgac ggcaactaca agacccgcgc
      961 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1021 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1081 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     1141 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1201 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga
     1261 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1321 tctcggcatg gacgagctgt acaagggctc tggtgctacc aacttctcac tgttgaaaca
     1381 ggcaggggat gtagaggaga atccagggcc tggtgctagt ggagactaca aagaccatga
     1441 cggagattat aaagatcatg acatcgatta caaggatgac gatgacaagt ccggactcag
     1501 atctggaggc tccggaggcc cagagccagc gaagtctgct cccgccccga aaaagggctc
     1561 caagaaggcg gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa
     1621 ggagagctat tccatctatg tgtataaggt tctgaagcag gtccaccctg acaccggcat
     1681 ttcgtccaag gccatgggca tcatgaactc gtttgtgaac gacattttcg agcgcatcgc
     1741 aggtgaggct tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat
     1801 ccagacggcc gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg
     1861 tactaaggcc atcaccaagt acaccagcgc taagtaatga ggcgcgccgc ggccgcgaat
     1921 tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     1981 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2041 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac
     2101 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2161 tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg
     2221 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2281 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2341 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac
     2401 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     2461 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     2521 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctatc agcacacaat
     2581 tgcccattat acgcgcgtat aatggactat tgtgtgctga tatgatgcgg tattttctcc
     2641 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     2701 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2761 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2821 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2881 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     2941 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3001 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     3061 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3121 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     3181 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3241 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3301 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3361 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3421 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3481 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3541 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3601 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3661 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3721 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3781 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3841 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3901 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3961 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4021 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4081 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4141 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4201 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4261 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4321 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4381 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4441 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4501 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4561 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4621 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4681 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4741 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4801 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4861 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4921 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4981 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5041 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5101 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5161 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcata tcagcacaca
     5221 attgcccatt atacgcgcgt ataatggact attgtgtgct gataacatgt
//
