LOCUS       sequence-466274-        4758 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2275m 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 4758)
  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 4758)
  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: AiP1608 - pAAV-AiE2275m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(584..1375,1376..1444))
                     /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     1207..1226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1445..1549)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1617..1635
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1673..1695)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1795..1814
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1831..2286)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1968..1989)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2180..2199
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2361..2501
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2372..2501
                     /label=AAV2 ITR
     polyA_signal    2619..2730
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2917..2937
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3030..3740,3741..3743,3744..3746))
                     /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(3072..3093)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3421..3440
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3680..3701
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3743..3752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   4392..4521
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(4583..4758,1..413))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4663..4682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
       61 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      121 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      181 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      241 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      301 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      361 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      421 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
      481 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
      541 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
      601 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
      661 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
      721 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
      781 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
      841 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
      901 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
      961 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     1021 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     1081 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     1141 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     1201 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     1261 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     1321 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     1381 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     1441 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     1501 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     1561 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     1621 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     1681 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     1741 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     1801 cagagcagat tgtactgaga gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat
     1861 tcgcgttaaa tttttgttaa atcagctcat tttttaacca atagaccgaa atcggcaaaa
     1921 tcccttataa atcaaaagaa tagcccgaga tagagttgag tgttgttcca gtttggaaca
     1981 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2041 gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta
     2101 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2161 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa
     2221 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2281 gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag
     2341 aaaataccgc atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     2401 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     2461 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgct cggtccgcac
     2521 gtgagatctc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc
     2581 caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt
     2641 aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa
     2701 caacagatgg ctggcaacta gaaggcacag tcgaagatct ctcgagccac ggaattgtca
     2761 gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc
     2821 gtggcaagaa ctaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca
     2881 atagcatgat acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag
     2941 ttaagaatac cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt atgatatcga
     3001 attcgcggcc gcggcgcgcc gtcgacttac ttgtacagct cgtccatgcc gagagtgatc
     3061 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     3121 ctcagcttgg actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     3181 atgggggtgt tctgctggta gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg
     3241 cggatcttga agttggcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg
     3301 tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     3361 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     3421 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc
     3481 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc
     3541 acgccgtagc ccagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     3601 cagatcagct tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg
     3661 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     3721 aacagctcct cgcccttgct caccatggtg gcgaccggta gcgctagctt cgaaagatct
     3781 ggatcccaga agcaaatgta agcaatagat ggctctgccc tgacttttat gcccagcccg
     3841 agctccgccg gcggtgagag gcaatactct gaagattgct tcatcagtat ctttccccag
     3901 aggaatgttg tatatcctgg gaaataagaa tatgggaagg tatttaggct tctaagaagg
     3961 ctgttgcact ttcctggtgc aaagagcctg cagtaagcag tccttgcatg ctctgccttg
     4021 agaatgcttt gcccttctat ccagaggggt gaggtattga atttgaatag gatggcaatt
     4081 ctgtggtgtt ctcggcctct ccattttagt gttgccagga atggctcctg tagctgggct
     4141 taaacccggc tgtgagaggc actggcgttt acagaatgag agctaagatt aaccatccac
     4201 gtgggaaaga tctgtgtgag gagatctgag gagacactgc ctccttactt ttgtggatag
     4261 tgttcaggac attcacatta caaaagaaaa gtctcaggac attttgaaaa caaacaatag
     4321 gaactttttt tttgtttgtt tgcttccaac tagccgccgg cgattattac tcttacaacg
     4381 cgtgcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     4441 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc
     4501 gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4561 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4621 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4681 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4741 ccgcctttct cccttcgg
//
