LOCUS       sequence-438976-        4565 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2394m 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 4565)
  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 4565)
  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: AiP1725 - pAAV-AiE2394m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(33..51)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        119..223
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(224..292,293..1084)
                     /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(442..461)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1255..1843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1744..1763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1905..2034
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2481..2490
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2487..2489,2490..2492,2493..3203)
                     /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(2532..2553)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2793..2812)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3140..3161
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3296..3316)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3503..3614
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3732..3872
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3732..3861
                     /label=AAV2 ITR
     rep_origin      3947..4402
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4034..4053)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4244..4265
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4419..4438)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4538..4560
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
       61 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
      121 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
      181 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
      241 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
      301 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
      361 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
      421 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
      481 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
      541 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
      601 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
      661 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
      721 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
      781 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
      841 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
      901 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
      961 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     1021 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     1081 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     1141 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     1201 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     1261 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1321 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1381 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     1441 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1501 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1561 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     1621 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     1681 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     1741 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     1801 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     1861 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg
     1921 cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     1981 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
     2041 gcacgcgttg taagagtaat aatcgtgaat gcaaagcagt atgtaataaa ttatgtgtct
     2101 ttttaagcaa tgaataaatg ttaagcaaac agatatgatg atactttttg attgctctct
     2161 ctctctctct ctttctcaaa aaaaaatccc agctccctct cactgtgttt gcttgccagg
     2221 aagcttcagt ggtgcctttg tgtaagcagt ttctggtgtt gagatttaaa aataaaacag
     2281 ggcttacttt ctagcacgct tgtcctttca aggatagagc tgtagctcaa acatccatcc
     2341 agagtctgga gggttcacta aggccttttc atttctccga gcgcacggag ctcgggctgg
     2401 gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat ccagatcttt
     2461 cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     2521 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     2581 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg
     2641 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc gtgcagtgct
     2701 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2761 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2821 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2881 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     2941 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     3001 tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     3061 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc
     3121 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3181 tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga attcgatatc
     3241 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3301 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     3361 gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac
     3421 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3481 ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3541 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3601 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3661 caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac
     3721 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3781 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     3841 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     3901 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     3961 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4021 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4081 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4141 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     4201 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4261 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     4321 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4381 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     4441 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4501 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4561 ttttc
//
