LOCUS       sequence-461969-        4695 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2663m 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 4695)
  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 4695)
  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: AiP2106 - pAAV-AiE2663m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4695
                     /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      707..716
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(713..715,716..718,719..1429)
                     /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(758..779)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1019..1038)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1366..1387
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1522..1542)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1729..1840
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1958..2098
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1958..2087
                     /label=AAV2 ITR
     rep_origin      2173..2628
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2260..2279)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2470..2491
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2645..2664)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2764..2786
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2824..2842)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2910..3014
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3015..3083,3084..3875)
                     /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(3233..3252)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4046..4634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4535..4554
                     /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 gcggccgcac gcgttctcct ttccttagtc tttccactct gctggcctac
      181 gggcagctgt gagtttttca tgttcctggt gctgctgcct cggtttctag catatgggag
      241 atggttctaa aaggtttagc tatcagtgga gtttgtagca tagccctgaa gtcacagaca
      301 atctaagcaa agtgaaaatt ttccatcaca tgacaaaaca aaaacaaaaa gaagaacact
      361 aaacactggt gtgatttcct gtgctctctg ggatactctt gccccatctg cttttaaatt
      421 tttgagttaa atccaaatat tcatagttct tgacagaaca cacttagaac aaacaaagtc
      481 tctactatga atctctgact gtttcagcat gcctttcttt ctcagtctgt taccataccc
      541 aaagcaaatt gaaaacagat ttgatatcca atctggggag acttctgcca tttctttatt
      601 ttcctcgccg gcggagctcg ggctgggcat aaaagtcagg gcagagccat ctattgctta
      661 catttgcttc tgggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag
      721 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      781 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      841 gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      901 caccctgggc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
      961 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1021 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1081 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1141 gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     1201 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
     1261 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1321 ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1381 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg
     1441 cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag
     1501 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     1561 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     1621 ctggttagtt cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     1681 ggctcggctg ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta
     1741 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1801 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1861 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     1921 gcaggcatga gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     1981 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     2041 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     2101 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     2161 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2221 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     2281 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2341 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     2401 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2461 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     2521 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     2581 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     2641 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     2701 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2761 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     2821 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     2881 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2941 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3001 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3061 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3121 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3181 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3241 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3301 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3361 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3421 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3481 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3541 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3601 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3661 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3721 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3781 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3841 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3901 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3961 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4021 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4081 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4141 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4201 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4261 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4321 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4381 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4441 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4501 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4561 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4621 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4681 cttttgctca catgt
//
