LOCUS       sequence-470580-        4923 bp    DNA     circular SYN 30-APR-2025
DEFINITION  AiE0597m 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 4923)
  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 4923)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 30, 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: AiP12942 - pAAV-AiE0597m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(510..1301,1302..1370))
                     /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     1133..1152
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1371..1475)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1543..1561
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1599..1621)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1721..1740
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1757..2212)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1894..1915)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2106..2125
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2287..2427
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2298..2427
                     /label=AAV2 ITR
     polyA_signal    2545..2656
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2843..2863
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2956..3666,3667..3669,3670..3672))
                     /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-C
                     /note="EGFP, forward primer"
     primer_bind     3347..3366
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3606..3627
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3669..3678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4483..4612
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4674..4923,1..339))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4754..4773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
       61 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      121 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      181 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      241 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
      301 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
      361 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
      421 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
      481 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
      541 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      601 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
      661 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
      721 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
      781 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
      841 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
      901 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
      961 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1021 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1081 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1141 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1201 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1261 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1321 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1381 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1441 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1501 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     1561 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     1621 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     1681 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     1741 ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     1801 tgttaaatca gctcattttt taaccaatag accgaaatcg gcaaaatccc ttataaatca
     1861 aaagaatagc ccgagataga gttgagtgtt gttccagttt ggaacaagag tccactatta
     1921 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     1981 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2041 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2101 aaggaaggga agaaagcgaa aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg
     2161 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt
     2221 tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     2281 ggcgcccctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
     2341 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     2401 tggccaactc catcactagg ggttcctgcg gccgctcggt ccgcacgtga gatctcatgc
     2461 ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata
     2521 gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg
     2581 ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg
     2641 caactagaag gcacagtcga agatctctcg agccacggaa ttgtcagtgc ccaacagccg
     2701 agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caagaactaa
     2761 ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa
     2821 aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt
     2881 caatctttca caaattttgt aatccagagg ttgattatga tatcgaattc gcggccgcgg
     2941 cgcgccgtcg acttacttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac
     3001 gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gcttggactg
     3061 gtagctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg
     3121 ctggtagtgg tcggcgagct gcacgccgcc gtcctcgatg ttgtggcgga tcttgaagtt
     3181 ggccttgatg ccgttcttct gcttgtcggc ggtgatatag acgttgtggc tgttgtagtt
     3241 gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc
     3301 gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc
     3361 gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa
     3421 gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag cactgcacgc cgtagcccag
     3481 ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga tcagcttcag
     3541 ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc
     3601 gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc
     3661 cttgctcacc atggtggcga ccggtagcgc tagcttcgaa agatctggat cccagaagca
     3721 aatgtaagca atagatggct ctgccctgac ttttatgccc agcccgagct ctgtccatac
     3781 cagggttcgt cttcccatct caaatgaacc ctgcctagaa actccctcac aggcatgacc
     3841 agaagcttgt cacctaagta attttagatc ctatcaggtt cacagcaagc aaggtaattg
     3901 tataagtcag gttcaaacaa cgtatcattt gtcaatcact gatactttaa aacctcattt
     3961 tctttgtgtt tttataaggc attcattcat tcattcaata cacgtgttta agtgtctaga
     4021 gctgtgatgt tcagtacagt aggaagtaac cacatataga tacttaaatt taagtgaatt
     4081 aaaattaaac ttaaaatttt aaatttagta tcttagtcac actagcctca tttcaactgc
     4141 tcaaataggc atcacatatg gtcagcagct gccatcttgg atgaagtgaa tataaaatac
     4201 tttaatcatc agaaaaactt ttactagaat gttctggcca ttctgccaca ttattacatt
     4261 ctagggatac atcatgtgaa taggtacagc cccttgaatt gtgaattata tacacaattc
     4321 aatatagtag acaataatga atgtaataag gcaaacacaa tgctttgtaa agagcattga
     4381 gaagcaccat tgtatacaac tgaaataatg tgtagactct aaaagaaaaa gtgctggagg
     4441 cagaaagaac aggaggtaca agagctctac gcgtgcggcc gcaggaaccc ctagtgatgg
     4501 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4561 cccgacgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggacatgtga
     4621 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4681 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4741 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4801 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4861 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4921 ggc
//
