LOCUS       sequence-432543-        4733 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2343m 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 4733)
  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 4733)
  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: AiP1661 - pAAV-AiE2343m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(152..171)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     271..293
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(331..349)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        417..521
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(522..590,591..1382)
                     /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(740..759)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1553..2141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2042..2061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2203..2332
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2947..2956
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2953..2955,2956..2958,2959..3669)
                     /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(2998..3019)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3259..3278)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3606..3627
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3762..3782)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3969..4080
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4198..4338
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4198..4327
                     /label=AAV2 ITR
     rep_origin      join(4413..4733,1..135)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4500..4519)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4710..4731
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
       61 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
      121 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
      181 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
      241 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
      301 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
      361 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
      421 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      481 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
      541 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
      601 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
      661 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
      721 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
      781 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
      841 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
      901 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
      961 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     1021 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     1081 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     1141 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     1201 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     1261 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     1321 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     1381 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     1441 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     1501 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1561 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1621 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1681 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1741 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1801 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1861 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1921 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1981 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2041 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2101 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2161 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg
     2221 ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
     2281 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
     2341 acgcgttgta agagtaataa tcgttaacca caagtaagaa gattttcctt tcatttcctc
     2401 attcctaagt attcccaagc cgacaagtac cttgtcatta tgtggctgct ccagccacgc
     2461 ttacgggttt tgagagcaat gtgattagat agtggcataa acttatgaga tgtttagcat
     2521 atgacatcat ctggtggtat ttgcctctca tttggcctga ctagagagag caatgcttac
     2581 tgcagtgaaa tgcctttctg ttggcagaat ttcagcatca gggagcattg ttggtagccc
     2641 tttagtagct cctgcctact ttacatacat cgatgaacca accaaaaact attatgaaat
     2701 ggaatggttt tggtaagaaa tcttgttatg attaagagaa acaaacaaaa tgttaaaaag
     2761 cggtctggga aagcttcatg tggaccatga gatatgaccc aatttttcaa ggcagattaa
     2821 aataaagaca gaggaaagaa aggaaggaag gcggagctcg ggctgggcat aaaagtcagg
     2881 gcagagccat ctattgctta catttgcttc tgggatccag atctttcgaa gctagcgcta
     2941 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3001 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     3061 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     3121 ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc
     3181 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3241 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3301 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3361 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     3421 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     3481 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3541 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     3601 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3661 ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg
     3721 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3781 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3841 ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc
     3901 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggctcgagag
     3961 atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4021 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4081 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4141 ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag cggccgcagg
     4201 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     4261 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     4321 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4381 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     4441 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     4501 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4561 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4621 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4681 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaa
//
