LOCUS       sequence-466327-        4763 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE2336m 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 4763)
  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 4763)
  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: AiP1910 - pAAV-AiE2336m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   244..373
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(435..1023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(515..534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1194..1985,1986..2054))
                     /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     1817..1836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2055..2159)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2227..2245
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2283..2305)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2405..2424
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2441..2896)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2578..2599)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2790..2809
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2971..3111
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2982..3111
                     /label=AAV2 ITR
     polyA_signal    3229..3340
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3527..3547
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3640..4350,4351..4353,4354..4356))
                     /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(3682..3703)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4031..4050
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4290..4311
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4353..4362
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cctccaccat gctcgacagg aagtactaga gtctgtggca tcagtgctgg tgaaatgtta
       61 ataataataa gaggccacat cttgaggata tcgcaaaggg tcccgttgga gagcagagct
      121 ccagtccgac accacagtaa caaaaggcct ttctccctag atgctagcaa aggtggggta
      181 gagaacctga accaactccc acatcacaga aacagaactt gcctcgataa tgggtgcggc
      241 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
      301 aggccgggcg accaaaggtc gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc
      361 agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      421 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      481 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      541 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      601 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      661 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      721 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
      781 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
      841 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
      901 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
      961 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1021 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1081 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1141 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1201 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1261 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1321 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1381 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1441 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1501 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1561 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1621 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1681 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1741 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1801 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1861 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1921 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     1981 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2041 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     2101 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2161 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     2221 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     2281 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     2341 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     2401 tatgcggcat cagagcagat tgtactgaga gtgcaccata aaattgtaaa cgttaatatt
     2461 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca atagaccgaa
     2521 atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagagttgag tgttgttcca
     2581 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     2641 gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttggggtcg
     2701 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     2761 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag
     2821 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     2881 ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat
     2941 gcgtaaggag aaaataccgc atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca
     3001 ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     3061 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgct
     3121 cggtccgcac gtgagatctc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc
     3181 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc
     3241 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg
     3301 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaagatct ctcgagccac
     3361 ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc aggcggcgat
     3421 gagttccgcc gtggcaagaa ctaaccagga tttatacaag gaggagaaaa tgaaagccat
     3481 acgggaagca atagcatgat acaaaggcat taaagcagcg tatccacata gcgtaaaagg
     3541 agcaacatag ttaagaatac cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt
     3601 atgatatcga attcgcggcc gcggcgcgcc gtcgacttac ttgtacagct cgtccatgcc
     3661 gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt
     3721 ggggtctttg ctcagcttgg actggtagct caggtagtgg ttgtcgggca gcagcacggg
     3781 gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc cgccgtcctc
     3841 gatgttgtgg cggatcttga agttggcctt gatgccgttc ttctgcttgt cggcggtgat
     3901 atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc
     3961 cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac
     4021 ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta
     4081 gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc
     4141 gaagcactgc acgccgtagc ccagggtggt cacgagggtg ggccagggca cgggcagctt
     4201 gccggtggtg cagatcagct tcagggtcag cttgccgtag gtggcatcgc cctcgccctc
     4261 gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac
     4321 caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggta gcgctagctt
     4381 cgaaagatct ggatcccaga agcaaatgta agcaatagat ggctctgccc tgacttttat
     4441 gcccagcccg agctccgaca tttgaagaaa aaaaatatgt tactaaccat tttactccat
     4501 aaaaacacca cagacttcaa agagcatgtt cagttgccct gcccacacaa caaaggccaa
     4561 gaggagacag acccattcca ggctgtgagg aaacactcat tctactctct tcttggggcc
     4621 acattaaaga ctacagttga gaactgtaac ttctgccctg aatgcctccc acgccactgt
     4681 cagtaaaacc atgctggcag cctacaatca ggcaatatag tctgcctgct agaatatcca
     4741 tagaggccca ctgggagtca gga
//
