LOCUS       sequence-462017-        4964 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2496m 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 4964)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, 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 J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 4964)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP1985 - pAAV-AiE2496m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4964
                     /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      976..985
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(982..984,985..987,988..1698)
                     /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(1027..1048)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1288..1307)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1635..1656
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1791..1811)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1998..2109
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2227..2367
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2227..2356
                     /label=AAV2 ITR
     rep_origin      2442..2897
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2529..2548)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2739..2760
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2914..2933)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3033..3055
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3093..3111)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3179..3283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3284..3352,3353..4144)
                     /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(3502..3521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4315..4903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4804..4823
                     /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 gcggccgcaa gcatcagcgt gggggcaggg cgggagactc atataaaacg
      181 cacaaagatg caaagttggt tgtgggtctc gggggtgggc ccaggtatta ttgattagta
      241 actgctaggg gatttgtgtg cacagctgag aggacagcat cccagagccc cccaaagctg
      301 catggacatt gagcactggg caaaaggctg tgttcttttc atgcttccga tttgtactga
      361 aggccgtctt tgctctatag cttgtttttg caatacatgg tggaagaatt tggatggtgt
      421 gttcttggcg ctggagtttt agctcgtagg gtatcggtta catattcatc tggaaattta
      481 caagctccag ttgaatatca aaattcacac gaagcgagcc aagcaaagga gctgctctca
      541 cttcctaata acagtttttc aaagccaaac attgctcagg ccttcagctt gcttgcttga
      601 atcataacag ctgggctccg gggcatttga ggaacctgac ctaccctaac tacgagtgta
      661 aaatatcgct tttttgtttt gttttgtttg tttgtttgtt ttttgttttt cgagacaggg
      721 tttctctgta tagccctggc tgtcctggaa ctcactttgt agaccaggct gtcctggaac
      781 tcagaaatcc gaaaatatcg cttttgttgt aggtaccaca ccaaggaggc cggctgccag
      841 tgggcagctg cgctcttgca tcctgggaag gtcccatggc cggagctcgg gctgggcata
      901 aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccaga tctttcgaag
      961 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1021 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1081 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
     1141 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc
     1201 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1261 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1321 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1381 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1441 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     1501 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1561 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
     1621 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1681 atggacgagc tgtacaagta agtcgacggc gcgccgcggc cgcgaattcg atatcataat
     1741 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     1801 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     1861 gctttcattt tctcctcctt gtataaatcc tggttagttc ttgccacggc ggaactcatc
     1921 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     1981 gctcgagaga tcttcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2041 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2101 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2161 cagcaagggg gaggattggg aagacaatag caggcatgag atctcacgtg cggaccgagc
     2221 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     2281 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     2341 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     2401 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     2461 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     2521 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2581 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2641 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2701 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2761 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     2821 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     2881 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     2941 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3001 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3061 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3121 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3181 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3241 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3301 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3361 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3421 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3481 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3541 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3601 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3661 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3721 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3781 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3841 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3901 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3961 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4021 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4081 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4141 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4201 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4261 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4321 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4381 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4441 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4501 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4561 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4621 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4681 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4741 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4801 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4861 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4921 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
