LOCUS       sequence-466332-        4815 bp DNA     circular SYN 24-MAR-2025
DEFINITION  AiE2526m 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 4815)
  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 4815)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 24, 2025 from SnapGene Server 7.0.3
            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."
FEATURES             Location/Qualifiers
     source          1..4815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     104..123
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   265..394
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1091..1100
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1097..1813
                     /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(1142..1163)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1403..1422)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1750..1771
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1906..1926)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2113..2224
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2342..2482
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2342..2471
                     /label=AAV2 ITR
     rep_origin      2557..3012
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2644..2663)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2854..2875
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3029..3048)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3148..3170
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3208..3226)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3294..3398
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3399..4259
                     /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(3617..3636)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4430..4815,1..203)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
       61 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      121 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
      181 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
      241 tttgctggcc ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg
      301 ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
      361 gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgttg taagagtaat
      421 aatcgaatct gtccaagatt cagagagata attacacttt cactgcattg ccaggggtca
      481 gcgtctctgt ggaccggagg agttgattta ctaacaatag tgttacattt aaaaaaaaaa
      541 tggcgtttga gattataatg ttaatggctg tggtcgcagg ggaatgctgc cgcttccaac
      601 taattccaca caacttcttt ttcctccttc ctttcttcct gggcttcatc tttgtaagga
      661 actggagcct tcctaatgtt taatttgctg caggcgccag aaggttatga ccagctctgg
      721 ggcttggtca tgactggttg ttccggtagt gtttgcattg tggcctcgac aggctgtctt
      781 gtcaacaaca aggaccagag tgaaggacag tgctgcgagc acctctcagc gaggcgagca
      841 gagagcaccc aggcagaagg cagcactggc tgggcttctc cagctttcga ttctctccgg
      901 ccaggtttgc agcctggaga aacatgccac caggctggat ctctgacttc cttggtcctg
      961 gagctcttca gtcaaaccta gacctgccct gttctcggag ctcgggctgg gcataaaagt
     1021 cagggcagag ccatctattg cttacatttg cttctgggat ccagatcttt cgaagctagc
     1081 gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1141 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1201 gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc
     1261 cgtgccctgg cccaccctcg tgaccaccct gggctacggc gtgcagtgct tcgcccgcta
     1321 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1381 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1441 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1501 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc
     1561 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg
     1621 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1681 gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa
     1741 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1801 cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga attcgatatc ataatcaacc
     1861 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1921 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1981 cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc
     2041 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggctcg
     2101 agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2161 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2221 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2281 agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac cgagcggccg
     2341 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     2401 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     2461 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     2521 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     2581 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     2641 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2701 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2761 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2821 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2881 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     2941 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3001 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3061 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3121 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3181 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3241 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3301 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3361 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3421 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3481 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3541 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3601 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3661 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3721 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3781 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3841 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3901 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3961 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4021 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4081 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4141 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4201 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4261 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4321 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4381 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4441 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4501 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4561 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4621 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4681 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4741 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4801 tgagatacct acagc
//
