LOCUS       sequence-466209-        5143 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2010m 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 5143)
  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 5143)
  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: AiP1198 - pAAV-AiE2010m-minBG-iCre-HGHpA
FEATURES             Location/Qualifiers
     source          1..5143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(633..1424,1425..1493))
                     /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     1256..1275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1494..1598)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1666..1684
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1722..1744)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1844..1863
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1880..2335)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2017..2038)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2229..2248
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2410..2550
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2421..2550
                     /label=AAV2 ITR
     polyA_signal    2590..3066
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     2848..2867
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(4115..4135)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      4138..4147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4826..4955
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(5017..5143,1..462))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5097..5116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
       61 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      121 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      181 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      241 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      301 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      361 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      421 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      481 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      541 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      601 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
      661 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
      721 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
      781 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
      841 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
      901 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
      961 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1021 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1081 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1141 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1201 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1261 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1321 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1381 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1441 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1501 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1561 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1621 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     1681 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     1741 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     1801 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     1861 gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     1921 ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa
     1981 tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa gagtccacta
     2041 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2101 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2161 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2221 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc
     2281 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtactat
     2341 ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2401 tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
     2461 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     2521 gagtggccaa ctccatcact aggggttcct gcggccgctc ggtccgcacg tggttaccta
     2581 caaaatcaga aggacaggga agggagcagt ggttcacgcc tgtaatccca gcaatttggg
     2641 aggccaaggt gggtagatca cctgagatta ggagttggag accagcctgg ccaatatggt
     2701 gaaaccccgt ctctaccaaa aaaacaaaaa ttagctgagc ctggtcatgc atgcctggaa
     2761 tcccaacaac tcgggaggct gaggcaggag aatcgcttga acccaggagg cggagattgc
     2821 agtgagccaa gattgtgcca ctgcactcca gcttggttcc caatagaccc cgcaggccct
     2881 acaggttgtc ttcccaactt gccccttgct ccataccacc cccctccacc ccataatatt
     2941 atagaaggac acctagtcag acaaaatgat gcaacttaat tttattagga caaggctggt
     3001 gggcactgga gtggcaactt ccagggccag gagaggcact ggggaggggt cacagggatg
     3061 ccacccgtag atctctcgag cagccttagt ccccatcctc gagcagcctc accatggccc
     3121 cagtctcaga gtccaggttt ctgatgtagt tcatcacaat gttcacattg gtccagccac
     3181 cagcctgcat gatttcaggg atggacacac cagccctggc catgtccctg gcagcaccca
     3241 ctctggcaga gtggccagac caggccaggt atctctgccc agagtcatcc ttggcaccat
     3301 agatcaggcg gtgggtggcc tcaaagatcc cttccagggc ccgggtggac agttgggagg
     3361 tggcagaagg ggcagccaca ccattctttc tgacccggca gaacaggtag ttgttggggt
     3421 catcagccac accagacaca gagatccatc tctccaccag cttggtaacc cccagggaca
     3481 gggccttctc cacaccagct gtggacacca gggtcttggt cctgccaatg tggatcagca
     3541 ttctcccacc atcggtgcgg gagatgtcct tcactctgat tctggcaatt tcggcaatgc
     3601 gcagcagggt gttgtaggca atgcccagga aggccaggtt cctgatgtcc tggcatctgt
     3661 cagagttctc catcagggat ctgacttggt caaagtcagt gcgttcaaag gccagggcct
     3721 gcttggctct ctccccagca tccacattct cctttctgat tctcctcatc accagggaca
     3781 cagcattgga gtcagaaggg cgaggcaggc cagatctcct gtgcagcatg ttgagctggc
     3841 ccaggtgctg ttggatggtc ttcacagcca ggcctctggc ttgcaggtac aggaggtagt
     3901 ccctcacatc ctcaggttca gcagggaacc atttcctgtt gttcagcttg caccaggcag
     3961 cccaggatct gcacacagac aggagcatct tccaggtgtg ttcagagaag gcctgcctgt
     4021 ccctgaacat gtccatcagg ttcttcctga cttcatcaga ggtggcatcc acagggaggg
     4081 caggcaggtt ttggtgcaca gtcagcaggt tggagacttt cctcttcttc ttgggcacca
     4141 tggtggcgga tcccagaagc aaatgtaagc aatagatggc tctgccctga cttttatgcc
     4201 cagcccgagc tctgttgcta cactagactc aatggagata aagaacactg tctgtttctg
     4261 gtttctgaaa atgcagcaga tatatctaca agtctggatg aaagtgatgg ttcacacttc
     4321 tcccttccca tttggaatct acttatgctt actgctcttt aagatgccta tgagggagtt
     4381 gttagtcata gaaagactca agataactaa aaggagataa agctattatt gctttacaca
     4441 gaactttgag tatgaatttt atcattttta taaattcttc taatattaca ctttgataag
     4501 ctgatttaca accatgtata ggcacatact ttgttcacaa tttaactcac ctcatctaaa
     4561 ttattttata ttgtgtccaa atagggagag aagaaaatgg gaaggaggaa gagatggagg
     4621 gacggaggga ggaagagaga gagagagaga tagagaagga aggagaaatc caagaggtgg
     4681 tttgcctttc caaaaagtca aggcatagtg tgaaaaacta taacattgta atgatgctat
     4741 gcacagtggt atatgtctgt gattccagca cttggaaggt aaaaacagga ggatcagagt
     4801 aaaagacaca tacgcgtgcg gccgcaggaa cccctagtga tggagttggc cactccctct
     4861 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc
     4921 ctcagtgagc gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa
     4981 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5041 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5101 ataccaggcg tttccccctg gaagctccct cgtgcgctct cct
//
