LOCUS       sequence-466199-        5706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2004m 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 5706)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle TL, Dwivedi D, Way SW, 
            Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya
            X, Roth JR, Alexander JR, Allen S, Amster A, Arbuckle J, Ayala A, 
            Baker PM, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D,
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Clark M, 
            Colbert K, Collman F, Daniel S, Departee M, DiValentin P, Donadio N,
            Dotson N, Egdorf T, Fliss T, Gabitto M, Garcia J, Gary A, Gasperini 
            M, Gloe J, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, 
            Halterman C, Haradon Z, Hastings SD, Helback O, Ho W, Hockemeyer D, 
            Huang C, Huff S, Hunker A, Johansen N, Jones D, Juneau Z, Kalmbach 
            B, Kannan M, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, Lee AY, 
            Leibly M, Lenz GH, Li S, Liang E, Lusk N, Madigan Z, Malloy J, 
            Malone J, McCue R, Melchor J, Mollenkopf T, Moosman S, Morin E, 
            Newman D, Ng L, Ngo K, Omstead V, Otto S, Oyama A, Pena N, Pham T, 
            Phillips E, Pom CA, Potekhina L, Ransford S, Ray PL, Rette D, 
            Reynoldson C, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sawyer L, 
            Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shepard N, Shulga L, 
            Sigler AR, Siverts L, Soliman S, Somasundaram S, Staats B, Stewart 
            K, Szelenyi E, Tieu M, Trader C, Tran A, van Velthoven CTJ, Walker 
            M, Wang Y, 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 JT, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   Cell. 2025 May 16:S0092-8674(25)00513-6. doi: 
            10.1016/j.cell.2025.05.002.
  PUBMED    40403729
REFERENCE   2  (bases 1 to 5706)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2025 
            May 16:S0092-8674(25)00513-6. doi: 10.1016/j.cell.2025.05.002."
COMMENT     Sequence Label: AiP982 - pAAV-AiE2004m-minBG-IRES2-iCre-HGHpA
FEATURES             Location/Qualifiers
     source          1..5706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(155..743)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(235..254)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(914..1705,1706..1774))
                     /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     1537..1556
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1775..1879)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1947..1965
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2003..2025)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2125..2144
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2161..2616)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2298..2319)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2510..2529
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2691..2831
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2702..2831
                     /label=AAV2 ITR
     polyA_signal    2871..3347
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3129..3148
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(4412..4432)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    4456..5008
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4458..5031
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4618..4637)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     4847..4864
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     repeat_region   join(5670..5706,1..93)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct
       61 cagtgagcga gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg
      121 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      181 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      241 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      301 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      361 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      421 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      481 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      541 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
      601 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      661 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      721 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
      781 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
      841 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
      901 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
      961 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1021 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1081 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1141 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1201 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1261 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1321 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1381 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1441 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1501 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1561 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1621 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1681 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1741 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     1801 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1861 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     1921 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     1981 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2041 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2101 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2161 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2221 tttttaacca atagaccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga
     2281 tagagttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2341 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca
     2401 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2461 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     2521 cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     2581 cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg
     2641 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc cctgcaggca
     2701 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
     2761 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     2821 taggggttcc tgcggccgct cggtccgcac gtggttacct acaaaatcag aaggacaggg
     2881 aagggagcag tggttcacgc ctgtaatccc agcaatttgg gaggccaagg tgggtagatc
     2941 acctgagatt aggagttgga gaccagcctg gccaatatgg tgaaaccccg tctctaccaa
     3001 aaaaacaaaa attagctgag cctggtcatg catgcctgga atcccaacaa ctcgggaggc
     3061 tgaggcagga gaatcgcttg aacccaggag gcggagattg cagtgagcca agattgtgcc
     3121 actgcactcc agcttggttc ccaatagacc ccgcaggccc tacaggttgt cttcccaact
     3181 tgccccttgc tccataccac ccccctccac cccataatat tatagaagga cacctagtca
     3241 gacaaaatga tgcaacttaa ttttattagg acaaggctgg tgggcactgg agtggcaact
     3301 tccagggcca ggagaggcac tggggagggg tcacagggat gccacccgta gatctctcga
     3361 gcagcggccg cggcgcgcct cattagtccc catcctcgag cagcctcacc atggccccag
     3421 tctcagagtc caggtttctg atgtagttca tcacaatgtt cacattggtc cagccaccag
     3481 cctgcatgat ttcagggatg gacacaccag ccctggccat gtccctggca gcacccactc
     3541 tggcagagtg gccagaccag gccaggtatc tctgcccaga gtcatccttg gcaccataga
     3601 tcaggcggtg ggtggcctca aagatccctt ccagggcccg ggtggacagt tgggaggtgg
     3661 cagaaggggc agccacacca ttctttctga cccggcagaa caggtagttg ttggggtcat
     3721 cagccacacc agacacagag atccatctct ccaccagctt ggtaaccccc agggacaggg
     3781 ccttctccac accagctgtg gacaccaggg tcttggtcct gccaatgtgg atcagcattc
     3841 tcccaccatc ggtgcgggag atgtccttca ctctgattct ggcaatttcg gcaatgcgca
     3901 gcagggtgtt gtaggcaatg cccaggaagg ccaggttcct gatgtcctgg catctgtcag
     3961 agttctccat cagggatctg acttggtcaa agtcagtgcg ttcaaaggcc agggcctgct
     4021 tggctctctc cccagcatcc acattctcct ttctgattct cctcatcacc agggacacag
     4081 cattggagtc agaagggcga ggcaggccag atctcctgtg cagcatgttg agctggccca
     4141 ggtgctgttg gatggtcttc acagccaggc ctctggcttg caggtacagg aggtagtccc
     4201 tcacatcctc aggttcagca gggaaccatt tcctgttgtt cagcttgcac caggcagccc
     4261 aggatctgca cacagacagg agcatcttcc aggtgtgttc agagaaggcc tgcctgtccc
     4321 tgaacatgtc catcaggttc ttcctgactt catcagaggt ggcatccaca gggagggcag
     4381 gcaggttttg gtgcacagtc agcaggttgg agactttcct cttcttcttg ggcaccatgg
     4441 tgctagctgt ggccatatta tcatcgtgtt tttcaaagga aaaccacgtc cccgtggttc
     4501 ggggggccta gacgtttttt taacctcgac taaacacatg taaagcatgt gcaccgaggc
     4561 cccagatcag atcccataca atggggtacc ttctgggcat ccttcagccc cttgttgaat
     4621 acgcttgagg agagccattt gactctttcc acaactatcc aactcacaac gtggcactgg
     4681 ggttgtgccg cctttgcagg tgtatcttat acacgtggct tttggccgca gaggcacctg
     4741 tcgccaggtg gggggttccg ctgcctgcaa agggtcgcta cagacgttgt ttgtcttcaa
     4801 gaagcttcca gaggaactgc ttccttcacg acattcaaca gaccttgcat tcctttggcg
     4861 agaggggaaa gacccctagg aatgctcgtc aagaagacag ggccaggttt ccgggccctc
     4921 acattgccaa aagacggcaa tatggtggaa aataacatat agacaaacgc acaccggcct
     4981 tattccaagc ggcttcggcc agtaacgtta gggggggggg agggagaggg gcggatccca
     5041 gaagcaaatg taagcaatag atggctctgc cctgactttt atgcccagcc cgagctcatt
     5101 tgtcctgtgc atagcattgc ctcttttcac taagttttct gtggccagtt gtgtctccag
     5161 ggtggaacag attgctgttg ctttagctga gcaccagatg aggtacttgg cttgcattga
     5221 ggggctcatg ttgaatgaaa attatgcttg tctctatttt tttggctccg cttgtagaac
     5281 tgaatgtggg cagattaaat caatgtacat atgatacaac tcaccataaa acacttgctc
     5341 aaacaatctt ttcgtgagcc aacaggaata gacaggaata aaaataaaca ccggtgccag
     5401 attacagagt tcctgatcag cacacgttga taggaaatta atgtgcatgt attgaaatag
     5461 cagaagatga tgtgttctga gatctctctg aaagcaagtt agcatggcaa tgcaagttag
     5521 tgagtcagtg atattcaaca gggtaacgct tccaagtgtg agtgttgaag cttttataaa
     5581 atgattgcat ttaaaatgta atttgtagca aatccaagga ctcaatgtaa tagtttgatt
     5641 gcccccaagg cttatacgcg tgcggccgca ggaaccccta gtgatggagt tggccactcc
     5701 ctctct
//
