LOCUS       sequence-462027-        4804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2613m 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 4804)
  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 4804)
  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: AiP2062 - pAAV-AiE2613m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4804
                     /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      816..825
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(822..824,825..827,828..1538)
                     /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(867..888)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1128..1147)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1475..1496
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1631..1651)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1838..1949
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2067..2207
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2067..2196
                     /label=AAV2 ITR
     rep_origin      2282..2737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2369..2388)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2579..2600
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2754..2773)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2873..2895
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2933..2951)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3019..3123
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3124..3192,3193..3984)
                     /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(3342..3361)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4155..4743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4644..4663
                     /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 gcgttgttcc ctgcccggag attaatttgt ttactccctt
      181 gcctggagat gaactgagag aactaatcgt ccatcagccg cacaaccttg ccccgcccgg
      241 ccctgaatgt ccattttctg gcaaaatatg tcctctctgc tgccaaggct ttttggaaaa
      301 tgctaatttt atcctcaaat taaaatttgt tcccctttcc tcattttaat aaaagcgagg
      361 catccggaaa tgttgccata gtgcagaagc gaacgttccc gacataattt tcaaaatgcc
      421 ttcgatcacg cggctttcgg aagggcctgt cctctggccc tgacctccct tccccatctc
      481 cacgccccct ggggcagcat ttacatttct taataacccc tcattaatat gcatgagaag
      541 gggtgggtaa taatggctga gcacgctgca ctaattcaat ttccacccgc ctcggcctgt
      601 ttacttaatc accaagggac ccgggttact cagcataatt atttattcgg aattagatct
      661 ggaggcatct ggactgagca aaatggaaac tggagtctct tggtcctgag ctctcgccgg
      721 cggagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct
      781 gggatccaga tctttcgaag ctagcgctac cggtcgccac catggtgagc aagggcgagg
      841 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      901 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc
      961 tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct
     1021 acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1081 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1141 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1201 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1261 acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     1321 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca
     1381 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca
     1441 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1501 ccgccgggat cactctcggc atggacgagc tgtacaagta agtcgacggc gcgccgcggc
     1561 cgcgaattcg atatcataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     1621 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     1681 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc
     1741 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     1801 tgggcactga caattccgtg gctcgagaga tcttcgactg tgccttctag ttgccagcca
     1861 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     1921 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     1981 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgag
     2041 atctcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     2101 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     2161 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     2221 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     2281 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2341 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2401 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2461 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     2521 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2581 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     2641 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     2701 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     2761 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     2821 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     2881 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     2941 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     3001 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3061 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3121 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3181 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3241 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3301 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3361 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3421 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3481 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3541 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3601 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3661 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3721 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3781 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3841 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3901 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3961 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4021 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4081 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4141 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4201 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4261 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4321 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4381 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4441 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4501 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4561 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4621 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4681 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4741 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4801 atgt
//
