LOCUS       sequence-461948-        5143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2016m 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 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 5143)
  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: AiP1300 - pAAV-AiE2016m-minBG-iCre-HGHpA
FEATURES             Location/Qualifiers
     source          1..5143
                     /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      809..818
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             821..841
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1890..2366
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2089..2108)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2406..2546
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2406..2535
                     /label=AAV2 ITR
     rep_origin      2621..3076
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2708..2727)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2918..2939
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3093..3112)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3212..3234
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3272..3290)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3358..3462
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3463..3531,3532..4323)
                     /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(3681..3700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4494..5082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4983..5002
                     /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 gcgtagggtg caggagaaat gtgacctcaa agtcttgttc
      181 tataactgtt ggaccttagg agagatctgt gctcagcaat ttaccaggac acccccaccc
      241 cacatgtctt gaccactgtc tggataactg gtatgcagga ccacactagg cttactcaca
      301 gtgtaaactc tcataaccat cactggagcc catcctgcct ggtagacaag gattcaacca
      361 tgactcattg tactttagtg gtgccatgct tagtcatcag gtgccctgtg ctctgacagc
      421 cgagggtcag agctggaatc acactcttgt tgtcttttaa tctctccctc cctttcttcc
      481 ttctttcttc actctgttgt gattgctcat ggaacagatc ctagctggtc tccctggcaa
      541 cctacatgat ttgagcccaa cagatggata atggggacat cgacttccaa tgtcattcaa
      601 cagaatcatt gccaagggag tctgatgagc aggcaactga gatgacaccc ttatcaatat
      661 agcttcattt tggcaatctg gagtaggtgt ttcaaaagga gagcccccac tgatgccagc
      721 aatacagaac gttcatgggc aaggagctcg ggctgggcat aaaagtcagg gcagagccat
      781 ctattgctta catttgcttc tgggatccgc caccatggtg cccaagaaga agaggaaagt
      841 ctccaacctg ctgactgtgc accaaaacct gcctgccctc cctgtggatg ccacctctga
      901 tgaagtcagg aagaacctga tggacatgtt cagggacagg caggccttct ctgaacacac
      961 ctggaagatg ctcctgtctg tgtgcagatc ctgggctgcc tggtgcaagc tgaacaacag
     1021 gaaatggttc cctgctgaac ctgaggatgt gagggactac ctcctgtacc tgcaagccag
     1081 aggcctggct gtgaagacca tccaacagca cctgggccag ctcaacatgc tgcacaggag
     1141 atctggcctg cctcgccctt ctgactccaa tgctgtgtcc ctggtgatga ggagaatcag
     1201 aaaggagaat gtggatgctg gggagagagc caagcaggcc ctggcctttg aacgcactga
     1261 ctttgaccaa gtcagatccc tgatggagaa ctctgacaga tgccaggaca tcaggaacct
     1321 ggccttcctg ggcattgcct acaacaccct gctgcgcatt gccgaaattg ccagaatcag
     1381 agtgaaggac atctcccgca ccgatggtgg gagaatgctg atccacattg gcaggaccaa
     1441 gaccctggtg tccacagctg gtgtggagaa ggccctgtcc ctgggggtta ccaagctggt
     1501 ggagagatgg atctctgtgt ctggtgtggc tgatgacccc aacaactacc tgttctgccg
     1561 ggtcagaaag aatggtgtgg ctgccccttc tgccacctcc caactgtcca cccgggccct
     1621 ggaagggatc tttgaggcca cccaccgcct gatctatggt gccaaggatg actctgggca
     1681 gagatacctg gcctggtctg gccactctgc cagagtgggt gctgccaggg acatggccag
     1741 ggctggtgtg tccatccctg aaatcatgca ggctggtggc tggaccaatg tgaacattgt
     1801 gatgaactac atcagaaacc tggactctga gactggggcc atggtgaggc tgctcgagga
     1861 tggggactaa ggctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc
     1921 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
     1981 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
     2041 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
     2101 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
     2161 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca
     2221 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
     2281 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
     2341 gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg
     2401 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     2461 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     2521 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     2581 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     2641 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     2701 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2761 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2821 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2881 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2941 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     3001 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3061 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3121 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3181 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3241 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3301 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     3361 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3421 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3481 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3541 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3601 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3661 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     3721 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3781 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3841 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3901 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3961 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4021 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4081 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4141 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4201 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4261 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4321 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4381 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4441 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4501 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4561 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4621 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4681 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4741 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4801 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4861 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4921 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4981 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5041 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5101 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
