LOCUS       sequence-462028-        5669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2619m 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 5669)
  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 5669)
  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: AiP2063 - pAAV-AiE2619m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5669
                     /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      1681..1690
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1687..1689,1690..1692,1693..2403)
                     /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(1732..1753)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1993..2012)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2340..2361
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2496..2516)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2703..2814
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2932..3072
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2932..3061
                     /label=AAV2 ITR
     rep_origin      3147..3602
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3234..3253)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3444..3465
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3619..3638)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3738..3760
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3798..3816)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3884..3988
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3989..4057,4058..4849)
                     /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(4207..4226)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5020..5608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5509..5528
                     /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 gcgtaacttt cctcctggct ccccataaag agggtagaaa
      181 caatataatt cgtttcttca tctggtcatg cattcccagt gaccacactc acagcaattt
      241 aaattagtca actgggcttt gacaaataga tgtattctca atgggcatct tcaaaattcc
      301 agtggagaga aaatcgacta ggcctaaaac tataagctta atggaatttg ctgagaatac
      361 gcagcagttt tcttcggttc aggccaacat gttccttggt gttttatgca agagctgtta
      421 ctgcagtcac accgaaggga agattcaacg agaatgggaa attatgagaa atgcccacac
      481 ttcctatttc ctatatctat tttagttggt ggcaattaca atttactacg caacactcaa
      541 aatataaacg gcactatgtg aaaggatata acagtgacag atttgcagcc tcaaagcgtg
      601 acttcaaaat attggtgccg aaaaggtttc ctgttccgga aggcagttga gactacaaag
      661 cccacgcagg tgaaggcgct ggcaggagcg agtgctctgc gtaccctgga tgttgagcaa
      721 agctgtgcag ggtagatggg aggcgccgcc cggccccata tggagccgac cttccgtgcc
      781 aatcgcacta tgctatggca cacacagcga gctgagcgag gaaatcctgc gaagcaggga
      841 aactgagagc ataaggcccg gagttgaaaa tcaagaagga aagccaagag tttgagggaa
      901 atgcagaggc agggttatgt gaggagaaca tgccagtgtc ctgtctcttt agccctgtcc
      961 accctagtac tagtccttgg cctagtacta ggcttgaatc agcagggcct ggcatgcctc
     1021 tgaagtcagc tggtttcagc cagtaagaac cactgcagaa agcccgagat tgagcccgga
     1081 agaggtctgt ttcttccctc gctcttatct acagcatggc gcagttcacg accctcagtg
     1141 atgagatggt ctttaatgct cctccccgac atactccttg ctctgacgct gattctgaga
     1201 tggagaatgg cctcccacca gtctgagcac aagggtgtgg cgtcacccgc aggcttgttc
     1261 cccttaactc tgctcacttc tttgtattcc gtcatttcat taaatgctca ggaaaaagcc
     1321 tgtctgtgat cctgactcat acagagaatt acaaggtccc ctactcagtc tcaaggagca
     1381 tgattaagaa cacaattgaa agtctccagg gctgcaggga tagtgcaggg gggtggaggt
     1441 ggcagcgggg tttcagggaa gacaggctcc ttgttctctt gcacttgcac cagtcttcga
     1501 tcctttgagc atttgtactt taaccagtgc ctctggtgtt cctgcacact tgcagacagg
     1561 tgggaagcca gtgcccccac gccggcggag ctcgggctgg gcataaaagt cagggcagag
     1621 ccatctattg cttacatttg cttctgggat ccagatcttt cgaagctagc gctaccggtc
     1681 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     1741 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     1801 acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg
     1861 cccaccctcg tgaccaccct gggctacggc gtgcagtgct tcgcccgcta ccccgaccac
     1921 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1981 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2041 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2101 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
     2161 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     2221 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2281 aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     2341 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2401 aagtaagtcg acggcgcgcc gcggccgcga attcgatatc ataatcaacc tctggattac
     2461 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     2521 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2581 tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc
     2641 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggctcg agagatcttc
     2701 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2761 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2821 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2881 ttgggaagac aatagcaggc atgagatctc acgtgcggac cgagcggccg caggaacccc
     2941 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     3001 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     3061 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3121 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     3181 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     3241 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3301 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3361 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3421 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     3481 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     3541 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     3601 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     3661 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3721 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3781 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3841 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3901 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3961 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4021 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4081 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4141 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4201 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4261 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4321 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4381 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4441 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4501 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4561 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4621 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4681 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4741 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4801 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4861 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4921 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4981 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5041 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5101 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5161 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5221 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5281 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5341 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5401 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5461 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5521 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5581 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5641 ggccttttgc tggccttttg ctcacatgt
//
