LOCUS       sequence-461873-        5335 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE0945m 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 5335)
  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 5335)
  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: AiP13298 -
            pAAV-AiE0945m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5335
                     /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      801..810
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(807..809,810..812,813..1523)
                     /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(1113..1132)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1460..1481
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1533..1589
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1602..1667
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1698..2075
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2162..2182)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2369..2480
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2598..2738
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2598..2727
                     /label=AAV2 ITR
     rep_origin      2813..3268
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2900..2919)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3110..3131
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3285..3304)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3404..3426
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3464..3482)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3550..3654
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3655..3723,3724..4515)
                     /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(3873..3892)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4686..5274
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5175..5194
                     /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 gcgtaggtag gttcacattc aggtgcaaac tgaaactttt
      181 gtttccccca gttttcaagt catgaattat tgaatatttc atgtattcta cctttgccag
      241 aacacttctt ttcccacctg catttggaaa ggatggcagg tgagcttcct gcttgccttt
      301 catgaatcca tcttgttctt tcagtacaac tgtttttctt tttgcttttt tttaagtagc
      361 atccaagctc tggtgggcga tttaatgagc aaaatgtgcc tgctccttct ggatataaac
      421 agaagacatc actggctgca aagtatgata tgggggaaga gattattcag tcagtgtttt
      481 ccgcttctct tcttgtttgt ctttcttttc catctcctgt agattgccca gttggtgctg
      541 ggacctcttc attcacttcc aaccttatgg catggaggtg tgctgctgtt gcagctcttt
      601 ctctctagag taaaatctga gacagcaact tgataataga agagacccat ctacacagcg
      661 tgtgtttctg ataatgcata tttttttctg ccttacgttt tctcacagag ctcgggctgg
      721 gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat ccagatcttt
      781 cgaagctagc gctaccggtc gccaccatgg tctccaaggg cgaggagctg ttcaccgggg
      841 tggtgcccat cctggtcgag ctggacggcg atgtcaacgg ccacaagttc agcgtgtccg
      901 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg
      961 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc gtgcagtgct
     1021 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1081 gctacgtcca ggagcgcacc atcttcttca aagacgacgg caactacaag acccgcgccg
     1141 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1201 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1261 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     1321 tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1381 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc
     1441 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1501 tcggcatgga cgagctgtac aagggctctg gtgctaccaa cttctcactg ttgaaacagg
     1561 caggggatgt agaggagaat ccagggcctg gtgctagtgg agactacaaa gaccatgacg
     1621 gagattataa agatcatgac atcgattaca aggatgacga tgacaagtcc ggactcagat
     1681 ctggaggctc cggaggccca gagccagcga agtctgctcc cgccccgaaa aagggctcca
     1741 agaaggcggt gactaaggcg cagaagaaag gcggcaagaa gcgcaagcgc agccgcaagg
     1801 agagctattc catctatgtg tacaaggttc tgaagcaggt ccaccctgac accggcattt
     1861 cgtccaaggc catgggcatc atgaattcgt ttgtgaacga cattttcgag cgcatcgcag
     1921 gagaggcttc ccgcctggcg cattacaaca agcgctcgac catcacctcc cgggagatcc
     1981 agacggccgt gcgcctgctg ctgcctgggg agttggccaa gcacgccgtg tccgagggta
     2041 ctaaggccat caccaagtac accagcgcta agtaatgagg cgcgccgcgg ccgcgaattc
     2101 gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2161 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2221 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg
     2281 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     2341 acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc atctgttgtt
     2401 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2461 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2521 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga gatctcacgt
     2581 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     2641 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     2701 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     2761 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     2821 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2881 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2941 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3001 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     3061 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3121 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     3181 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3241 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     3301 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3361 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3421 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3481 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3541 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3601 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3661 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3721 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3781 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3841 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3901 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3961 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4021 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4081 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4141 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4201 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4261 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4321 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4381 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4441 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4501 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4561 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4621 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4681 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4741 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4801 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4861 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4921 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4981 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5041 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5101 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5161 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5221 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5281 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
