LOCUS       sequence-432531-        4615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2114m 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 4615)
  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 4615)
  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: AiP1332 - pAAV-AiE2114m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(114..133)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     233..255
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(293..311)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        379..483
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(484..552,553..1344)
                     /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(702..721)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1515..2103
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2004..2023
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2165..2294
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2791..2800
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2797..2799,2800..2802,2803..3513)
                     /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(2842..2863)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3103..3122)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3450..3471
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3606..3626)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3813..3924
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4042..4182
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4042..4171
                     /label=AAV2 ITR
     rep_origin      join(4257..4615,1..97)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4344..4363)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4554..4575
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
       61 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
      121 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
      181 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
      241 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
      301 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
      361 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
      421 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
      481 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
      541 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
      601 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
      661 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
      721 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
      781 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
      841 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
      901 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
      961 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     1021 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     1081 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     1141 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     1201 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     1261 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1321 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     1381 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     1441 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     1501 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1561 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     1621 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     1681 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     1741 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     1801 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     1861 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     1921 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     1981 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2041 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2101 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2161 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac
     2221 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
     2281 cactaggggt tcctgcggcc gcacgcgttt ggcatttgac atttacctga agtctgtctt
     2341 gtgccgggtc ctgtccccag gttgggtcta agtctcacgc tggatctcag tgctcacagc
     2401 ttcctccaga acatacctgc tctggcatta tctttcctga gcagatgcta gaacttggct
     2461 cagagtgatg aagaggaagt gtgtcctctg gctaaagtcc aggggttgca gaggggagtg
     2521 tgtgagatat tttaatacat acctatcttg agaaggtagc aggggtgtta aagcatatgt
     2581 aatcatccag gcttcttcat atctcaagaa atctagcata gcacaaagat gggctagatg
     2641 tacccaaagg tgcatgcaaa acgtaagcag cttcttgttt cctatttcac tgagtttgag
     2701 ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat
     2761 ccagatcttt cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg
     2821 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2881 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc
     2941 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc
     3001 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     3061 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     3121 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     3181 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     3241 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     3301 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     3361 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg
     3421 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3481 gggatcactc tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga
     3541 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3601 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3661 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     3721 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3781 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     3841 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3901 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3961 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     4021 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     4081 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4141 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     4201 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     4261 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4321 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4381 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4441 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     4501 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4561 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg attta
//
