LOCUS       sequence-452588-        4962 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE0086m 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 4962)
  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 4962)
  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: AiP11580 -
            pAAV-hsA2-AiE0086m-minRho-SYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..4962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      1005..1014
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1011..1013,1014..1016,1017..1727)
                     /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(1056..1077)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1317..1336)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1664..1685
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1795..1815)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2002..2113
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2225..2365
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2225..2354
                     /label=AAV2 ITR
     rep_origin      2440..2895
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2527..2546)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2737..2758
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2912..2931)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3031..3053
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3091..3109)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3177..3281
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3282..3350,3351..4142)
                     /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(3500..3519)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4313..4901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4802..4821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca acgcgtttag aacaatggct ggcccatagt
      181 aaatgccgtg ttagtgtgtt agttgctgtt cttccacgtc agaagaggca cagacaaatt
      241 accaccaggt ggcgctcaga gtctgcggag gcatcacaac agccctgaat ttgaatcctg
      301 ctctgccact gcctagttga gaccttttac tacctgacta gctgtttgtg tattttaggt
      361 gtttgtttca ctctcatacc ccagcctcat ttcaaagcta gggatcatag cgccagcctt
      421 cagaaaagag actcaaaagg agcaaaaacg aagagtgcag ggctgctaga tagactccat
      481 ccagggaccg agcctgtgac gaatccagcc cctggcaaga gcccgacagg caggtgtaat
      541 cagcaaattt ctccggagac atcagagcag atccctcagc aagcacagct tctaaatttg
      601 tgctcaagaa tgattaatct gtttaagcaa attccttgag cataattaag atgccctaat
      661 caaggaagaa gattattatg cggcagcctt aggcactgat ttggtggcaa gagcccgtga
      721 ccacaaggcc tcaataactt ggtggggtag tgaaggtgac agagtggctt aggctgcagg
      781 gaggtgactg gggtgggggt ggggggaggt tagaattcga tatcataatc aaccataggt
      841 accgagctcg ggattcagcc gggagcttag ggaggggagg tcacttcata agggcctggg
      901 gggggagttg gagccacgag tcgtccagcc ggagccccgt gtggctgagc tccggcctca
      961 gaagcatccc cgggttggat ccttcgaagc tagcgctacc ggtcgccacc atggtgagca
     1021 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1081 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1141 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1201 ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1261 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1321 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1381 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1441 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1501 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     1561 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1621 accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1681 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacatca
     1741 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     1801 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     1861 tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact
     1921 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     1981 cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2041 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2101 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2161 aggacagcaa gggggaggat tgggaagaca atagcaggca tgcacgtgcg gaccgagcgg
     2221 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     2281 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     2341 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     2401 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     2461 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2521 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2581 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2641 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2701 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2761 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     2821 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2881 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     2941 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3001 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3061 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3121 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3181 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3241 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3301 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3361 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3421 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3481 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3541 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3601 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3661 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3721 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3781 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3841 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3901 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3961 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4021 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4081 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4141 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4201 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4261 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4321 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4381 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4441 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4501 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4561 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4621 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4681 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4741 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4801 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4861 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4921 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
