LOCUS       sequence-432536-        5247 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2165m 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 5247)
  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 5247)
  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: AiP1394 - pAAV-AiE2165m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   76..205
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1334..1343
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1340..1342,1343..1345,1346..2056)
                     /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(1385..1406)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1646..1665)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1993..2014
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2149..2169)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2356..2467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2585..2725
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2585..2714
                     /label=AAV2 ITR
     rep_origin      2800..3255
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2887..2906)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3097..3118
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3272..3291)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3391..3413
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3451..3469)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3537..3641
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3642..3710,3711..4502)
                     /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(3860..3879)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4673..5247,1..14)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5162..5181
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
       61 cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
      121 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      181 gccaactcca tcactagggg ttcctgcggc cgcacgcgta taatactgac tgctgcatga
      241 attcaaatat aatttcaaac atatggaggt tggagatttt ggaattaagg ttgagtatat
      301 aaaaatcatt aaaactttta aaattgtact catcaaaaat aactttgagc atcaagttgc
      361 tacaatcagt gtacaaatca gaaatataaa tgatattgta aaattatgca tgccttgaaa
      421 atggttacct taattttaat tttactatca gctgaataaa attttactgc atatttttct
      481 gttttttttt ctaatcaaaa taacccctcc cttgattcca aaacaataaa atgaaatgtc
      541 taaaatcatg ttattttatg ataacagttt tacagctgct aagatggaag catttctaac
      601 ctcccatatc tattataaat gacactgaaa ttatcatcct taggttgaaa aggaggttga
      661 aatatttgtc acaatacagt ctctccaccc caccatcctg agaagaatgc tatgactaaa
      721 attagagagt gcaggctgca ttacttaatt tcagtctttt gcagaaggag gagagcattg
      781 ggcctgagga gataaataga catcagaaca aaaagaagct aggcagttcc tgctgctttc
      841 ttcacaccct gggtgctaag cttatagtca tctaagaagc agatgctagg cttaaaaccc
      901 aattaaggtt gttggtaaat tgcactaaag cttacatttt gttttcttct agataaagaa
      961 gaaaaacaca attcatttat caaacacaag ccttgaattt tcctaaggtt tcactttaat
     1021 ttcacattcc ttaatgtgtt aaagtaaccc ttaaaatggt gtcaacccaa ttttacaata
     1081 ctaaattaac ttacatctca catgaaacat ccctacatat cagttaacag gacttgtctc
     1141 aaaatactta cctttaaaaa ttaatcatag catatcatca tacatttaaa atttcaacat
     1201 atcttttttt gttttgaaga atgatcttgg gttagtggaa gagctcgggc tgggcataaa
     1261 agtcagggca gagccatcta ttgcttacat ttgcttctgg gatccagatc tttcgaagct
     1321 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1381 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1441 cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct
     1501 gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg
     1561 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1621 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1681 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1741 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcac
     1801 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     1861 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1921 gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga
     1981 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2041 ggacgagctg tacaagtaag tcgacggcgc gccgcggccg cgaattcgat atcataatca
     2101 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2161 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2221 tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc
     2281 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggc
     2341 tcgagagatc ttcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2401 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2461 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2521 gcaaggggga ggattgggaa gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg
     2581 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     2641 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     2701 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     2761 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     2821 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     2881 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2941 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3001 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3061 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3121 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     3181 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3241 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3301 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3361 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3421 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3481 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3541 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3601 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3661 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3721 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3781 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3841 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3901 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3961 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4021 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4081 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4141 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4201 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4261 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4321 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4381 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4441 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4501 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4561 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4621 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4681 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4741 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4801 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4861 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4921 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4981 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5041 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5101 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5161 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5221 gatttttgtg atgctcgtca ggggggc
//
