LOCUS       sequence-466263-        5250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2164m 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 5250)
  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 5250)
  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: AiP1437 - pAAV-AiE2164m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     284..303
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   445..574
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1706..1715
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1712..1714,1715..1717,1718..2428)
                     /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(1757..1778)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2018..2037)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2365..2386
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2521..2541)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2728..2839
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2957..3097
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2957..3086
                     /label=AAV2 ITR
     rep_origin      3172..3627
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3259..3278)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3469..3490
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3644..3663)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3763..3785
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3823..3841)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3909..4013
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4014..4082,4083..4874)
                     /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(4232..4251)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5045..5250,1..383)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
       61 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      121 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      181 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      241 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      301 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
      361 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
      421 tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg
      481 ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
      541 gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgttg taagagtaat
      601 aatcgccggc gtgaagcccc cttcaaagga atttttcttc tctctaagca aatgcagcct
      661 tctttttttt tctgtgatct tactgttgaa aaactagaag ctaaagaaaa caaaatctaa
      721 acagaagcca catggtgtag tggagcaatg taatactata ctgtagaaat caaactgttg
      781 gcaataggtt gttttaatta ttaggctagt gtctcgattt taaatacacg ctttcatact
      841 taatattatg gtatgaggct gcggctctgt taccagccat gtgtttgctt tctcaacgct
      901 atgttcagac tatgagattt actaagtgcc ctccatcttt tcctgtggct tcctgcctac
      961 ttctttttgc ctctcagact gacacttcct ccctagcatc actttactcc aggagctaaa
     1021 agcccctccc agaactggaa gtgattctat tctggcaatt ctagcagaat tctcccaatg
     1081 ctagtagaat ttgcttcccc atctcccccc tgtgggacca gcttccactt tcagctccct
     1141 gggtttctcc ttcctctcgg cttgacatgt tcactctaac ccttcccagg attccagctc
     1201 acagttgcta gctgcttttt acagttggtg ccatcctatt tcatattcct gcttgccttc
     1261 tccaaagatc tagttaagta ttttacttac aactgctaac aactctcata caggattctc
     1321 agtattcaac tcactagacc catactaaaa aagaaattgt aaattgttca tttttttaaa
     1381 aaataaaaac acattttata gttcactaag cttattactg gacacaatag gaatatgggg
     1441 gaaaactatt tttaaatgat taattgtaat tcagggagac aaatttgcag acttgtgaat
     1501 ctggagtttc taatcatgcc tttattactc atagaaaata aaatttagat tactcatgta
     1561 aaataacacc ttctagtggt agtttgtttg ggggcaatac aaatcgccgg cggagctcgg
     1621 gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga
     1681 tctttcgaag ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
     1741 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1801 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac
     1861 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca
     1921 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1981 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2041 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2101 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2161 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     2221 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2281 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa
     2341 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2401 cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg
     2461 atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2521 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2581 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc
     2641 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2701 caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt
     2761 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2821 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2881 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg
     2941 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3001 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3061 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     3121 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3181 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3241 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3301 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3361 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3421 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3481 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3541 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3601 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3661 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3721 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3781 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3841 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3901 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3961 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4021 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4081 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4141 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4201 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4261 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4321 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4381 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4441 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4501 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4561 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4621 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4681 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4741 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4801 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4861 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4921 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4981 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5041 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5101 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5161 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5221 acttcaagaa ctctgtagca ccgcctacat
//
