LOCUS       sequence-470292-        5111 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 5111)
  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 5111)
  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: AiP1195 - pAAV-AiE2004m-minBG-iCre-HGHpA
FEATURES             Location/Qualifiers
     source          1..5111
                     /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      777..786
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             789..809
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1858..2334
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2057..2076)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2374..2514
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2374..2503
                     /label=AAV2 ITR
     rep_origin      2589..3044
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2676..2695)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2886..2907
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3061..3080)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3180..3202
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3240..3258)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3326..3430
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3431..3499,3500..4291)
                     /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(3649..3668)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4462..5050
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4951..4970
                     /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 gcgtggtacc agccttgggg gcaatcaaac tattacattg
      181 agtccttgga tttgctacaa attacatttt aaatgcaatc attttataaa agcttcaaca
      241 ctcacacttg gaagcgttac cctgttgaat atcactgact cactaacttg cattgccatg
      301 ctaacttgct ttcagagaga tctcagaaca catcatcttc tgctatttca atacatgcac
      361 attaatttcc tatcaacgtg tgctgatcag gaactctgta atctggcacc ggtgtttatt
      421 tttattcctg tctattcctg ttggctcacg aaaagattgt ttgagcaagt gttttatggt
      481 gagttgtatc atatgtacat tgatttaatc tgcccacatt cagttctaca agcggagcca
      541 aaaaaataga gacaagcata attttcattc aacatgagcc cctcaatgca agccaagtac
      601 ctcatctggt gctcagctaa agcaacagca atctgttcca ccctggagac acaactggcc
      661 acagaaaact tagtgaaaag aggcaatgct atgcacagga caaatcttaa ggagctcggg
      721 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccgcca
      781 ccatggtgcc caagaagaag aggaaagtct ccaacctgct gactgtgcac caaaacctgc
      841 ctgccctccc tgtggatgcc acctctgatg aagtcaggaa gaacctgatg gacatgttca
      901 gggacaggca ggccttctct gaacacacct ggaagatgct cctgtctgtg tgcagatcct
      961 gggctgcctg gtgcaagctg aacaacagga aatggttccc tgctgaacct gaggatgtga
     1021 gggactacct cctgtacctg caagccagag gcctggctgt gaagaccatc caacagcacc
     1081 tgggccagct caacatgctg cacaggagat ctggcctgcc tcgcccttct gactccaatg
     1141 ctgtgtccct ggtgatgagg agaatcagaa aggagaatgt ggatgctggg gagagagcca
     1201 agcaggccct ggcctttgaa cgcactgact ttgaccaagt cagatccctg atggagaact
     1261 ctgacagatg ccaggacatc aggaacctgg ccttcctggg cattgcctac aacaccctgc
     1321 tgcgcattgc cgaaattgcc agaatcagag tgaaggacat ctcccgcacc gatggtggga
     1381 gaatgctgat ccacattggc aggaccaaga ccctggtgtc cacagctggt gtggagaagg
     1441 ccctgtccct gggggttacc aagctggtgg agagatggat ctctgtgtct ggtgtggctg
     1501 atgaccccaa caactacctg ttctgccggg tcagaaagaa tggtgtggct gccccttctg
     1561 ccacctccca actgtccacc cgggccctgg aagggatctt tgaggccacc caccgcctga
     1621 tctatggtgc caaggatgac tctgggcaga gatacctggc ctggtctggc cactctgcca
     1681 gagtgggtgc tgccagggac atggccaggg ctggtgtgtc catccctgaa atcatgcagg
     1741 ctggtggctg gaccaatgtg aacattgtga tgaactacat cagaaacctg gactctgaga
     1801 ctggggccat ggtgaggctg ctcgaggatg gggactaagg ctgctcgaga gatctacggg
     1861 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     1921 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     1981 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     2041 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     2101 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     2161 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     2221 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     2281 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     2341 ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     2401 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     2461 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     2521 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     2581 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2641 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     2701 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2761 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     2821 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2881 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     2941 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     3001 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     3061 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     3121 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3181 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     3241 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     3301 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3361 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3421 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3481 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3541 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3601 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3661 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3721 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3781 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3841 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3901 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3961 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4021 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4081 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4141 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4201 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4261 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4321 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4381 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4441 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4501 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4561 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4621 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4681 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4741 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4801 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4861 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4921 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4981 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5041 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5101 tgctcacatg t
//
