LOCUS       sequence-459691-        5084 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE0456m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific brain cell types.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5084)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, Weed N, Wirthlin M, Wood 
            T, Wynalda B, Yao Z, Zhou T, Ariza J, Dee N, Reding M, Ronellenfitch
            K, Mufti S, Sunkin SM, Smith KA, Esposito L, Waters J, Thyagarajan 
            B, Yao S, Lein ES, Zeng H, Levi BP, Ngai J, Ting J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 5084)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP15290 - pAAV-AiE0456m-minBG-iCre(R297T)-BGH...
FEATURES             Location/Qualifiers
     source          1..5084
                     /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      1020..1029
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1032..1052
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2118..2229
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2347..2487
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2347..2476
                     /label=AAV2 ITR
     rep_origin      2562..3017
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2649..2668)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2859..2880
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3034..3053)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3153..3175
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3213..3231)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3299..3403
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3404..3472,3473..4264)
                     /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(3622..3641)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4435..5023
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4924..4943
                     /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 gcgttggtta gcactatttt gaatggacac ttctataaaa
      181 tttgttttaa aaacaatatg gaagagtcat aagatgtctg ggcacagtgc cccagtcaac
      241 tcatcaatgg gattctggaa tccaaaggtt atatggtgta aaaacacaga gagcaaaatg
      301 cacaaggatc ttcatagaac cataattttt aaaaagcaaa cacttccccc aaaggagcat
      361 gtaagacaac ttgaaggtat taaaatggga ttgtggtttt attagttaga gaacatcatg
      421 caaattatat tatctaggca ttataatgca aactttaaaa ggaaatgctc attatgtaat
      481 gttaagtgaa aaaaatcagg ccataaaatg aaatgtgcat tatgcatgca tatgatgaaa
      541 agacaagaaa tagataagaa tatgcagaaa tacaaatagt tggctgtggt gtgggcttct
      601 ggcttgtttt ctgttttaaa gccattacat catctttcca atgacaaact acattttaag
      661 ttattgaaaa caaaatctcg ttagccttgt ccttgcataa attcccagag aacacattat
      721 tcttgttccc ttctacctct gctacttgtc tttgatttgg acctgctcca cctcctctta
      781 cttttcagtc ctgaactaga tggaaaaaag aaatagaccc tcaggaaaca aggaagcaaa
      841 accacccaac tgtgtggaag ccgtgcattc aagctgcaaa cgagtggagt ggcttcaata
      901 ccttaactta tccccattgt tgcaaaggga gacccagaga gctcgggctg ggcataaaag
      961 tcagggcaga gccatctatt gcttacattt gcttctggga tccagatctt tcgaagctag
     1021 ccaccatggt gcccaagaag aagaggaaag tctccaacct gctgactgtg caccaaaacc
     1081 tgcctgccct ccctgtggat gccacctctg atgaagtcag gaagaacctg atggacatgt
     1141 tcagggacag gcaggccttc tctgaacaca cctggaagat gctcctgtct gtgtgcagat
     1201 cctgggctgc ctggtgcaag ctgaacaaca ggaaatggtt ccctgctgaa cctgaggatg
     1261 tgagggacta cctcctgtac ctgcaagcca gaggcctggc tgtgaagacc atccaacagc
     1321 acctgggcca gctcaacatg ctgcacagga gatctggcct gcctcgccct tctgactcca
     1381 atgctgtgtc cctggtgatg aggagaatca gaaaggagaa tgtggatgct ggggagagag
     1441 ccaagcaggc cctggccttt gaacgcactg actttgacca agtcagatcc ctgatggaga
     1501 actctgacag atgccaggac atcaggaacc tggccttcct gggcattgcc tacaacaccc
     1561 tgctgcgcat tgccgaaatt gccagaatca gagtgaagga catctcccgc accgatggtg
     1621 ggagaatgct gatccacatt ggcaggacca agaccctggt gtccacagct ggtgtggaga
     1681 aggccctgtc cctgggggtt accaagctgg tggagagatg gatctctgtg tctggtgtgg
     1741 ctgatgaccc caacaactac ctgttctgcc gggtcagaaa gaatggtgtg gctgcccctt
     1801 ctgccacctc ccaactgtcc acccgggccc tggaagggat ctttgaggcc acccaccgcc
     1861 tgatctatgg tgccaaggat gactctgggc agagatacct ggcctggtct ggccactctg
     1921 ccagagtggg tgctgccacc gacatggcca gggctggtgt gtccatccct gaaatcatgc
     1981 aggctggtgg ctggaccaat gtgaacattg tgatgaacta catcagaaac ctggactctg
     2041 agactggggc catggtgagg ctgctcgaag atggggactg aggcgcgccg aattcaagct
     2101 tctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2161 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2221 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2281 cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc
     2341 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     2401 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     2461 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     2581 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     2641 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2701 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2761 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2821 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2881 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     2941 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3001 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3061 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3121 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3181 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3241 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3301 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3361 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3421 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3481 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3541 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3601 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3661 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3721 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3781 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3841 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3901 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3961 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4021 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4081 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4141 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4201 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4261 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4321 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4381 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4441 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4501 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4561 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4621 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4681 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4741 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4801 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4861 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4921 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4981 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5041 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
