LOCUS       sequence-456270-        4825 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 4825)
  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 4825)
  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: AiP1307 - pAAV-AiE2016m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(90..109)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      903..1491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1392..1411
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1553..1682
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2389..2398
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2395..2397,2398..2400,2401..3111)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2440..2461)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2701..2720)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3048..3069
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3204..3224)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3411..3522
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3640..3780
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3640..3769
                     /label=AAV2 ITR
     rep_origin      3855..4310
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3942..3961)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4152..4173
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4327..4346)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4446..4468
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4506..4524)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4592..4696
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4697..4765,4766..4825,1..732)
                     /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"
ORIGIN
        1 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
       61 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
      121 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
      181 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
      241 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
      301 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
      361 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
      421 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
      481 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
      541 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
      601 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
      661 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
      721 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
      781 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
      841 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      901 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      961 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     1021 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     1081 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     1141 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1201 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     1261 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     1321 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     1381 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     1441 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     1501 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg
     1561 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc
     1621 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     1681 ctgcggccgc acgcgtaggg tgcaggagaa atgtgacctc aaagtcttgt tctataactg
     1741 ttggacctta ggagagatct gtgctcagca atttaccagg acacccccac cccacatgtc
     1801 ttgaccactg tctggataac tggtatgcag gaccacacta ggcttactca cagtgtaaac
     1861 tctcataacc atcactggag cccatcctgc ctggtagaca aggattcaac catgactcat
     1921 tgtactttag tggtgccatg cttagtcatc aggtgccctg tgctctgaca gccgagggtc
     1981 agagctggaa tcacactctt gttgtctttt aatctctccc tccctttctt ccttctttct
     2041 tcactctgtt gtgattgctc atggaacaga tcctagctgg tctccctggc aacctacatg
     2101 atttgagccc aacagatgga taatggggac atcgacttcc aatgtcattc aacagaatca
     2161 ttgccaaggg agtctgatga gcaggcaact gagatgacac ccttatcaat atagcttcat
     2221 tttggcaatc tggagtaggt gtttcaaaag gagagccccc actgatgcca gcaatacaga
     2281 acgttcatgg gcaaggagct cgggctgggc ataaaagtca gggcagagcc atctattgct
     2341 tacatttgct tctgggatcc agatctttcg aagctagcgc taccggtcgc caccatggtg
     2401 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2461 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2521 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2581 accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     2641 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2701 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2761 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2821 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
     2881 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
     2941 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3001 agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3061 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac
     3121 ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa
     3181 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3241 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3301 tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3361 ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc
     3421 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3481 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3541 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3601 tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     3661 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     3721 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     3781 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     3841 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3901 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     3961 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4021 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4081 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4141 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     4201 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     4261 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     4321 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4381 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4441 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4501 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4561 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4621 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4681 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4741 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4801 agatcagttg ggtgcacgag tgggt
//
