LOCUS       sequence-438960-        4714 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AiE2565m 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 4714)
  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 4714)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: AiP1861 - pAAV-AiE2565m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(29..97,98..889)
                     /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(247..266)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1060..1648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1549..1568
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1710..1839
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2435..2444
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2441..2443,2444..2446,2447..3157)
                     /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(2486..2507)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2747..2766)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3094..3115
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3250..3270)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3457..3568
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3686..3826
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3686..3815
                     /label=AAV2 ITR
     rep_origin      3901..4356
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3988..4007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4198..4219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4373..4392)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4492..4514
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4552..4570)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4638..4714,1..28)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
       61 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      121 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      181 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      241 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
      301 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      361 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      421 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      481 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      541 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
      601 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      661 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      721 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      781 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
      841 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
      901 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
      961 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1021 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1081 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1141 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1201 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1261 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1321 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1381 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1441 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1501 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1561 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1621 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1681 ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac
     1741 tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
     1801 gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgcacg cgttgtaaga
     1861 gtaataatcg ggaagcacct ctttgttaca taggctcagt tgcaattcag cagccagaag
     1921 gagccttttc cactggctgt taagagccgg gagggagggg gcaagcagaa gcttgacaga
     1981 ttctggtctt atctagaaaa aaaaaatcac tttacctatt agtttgagga aaatgggttg
     2041 gtgtgcttgc ccaaaccttt gatgccagcc actcctttta aaatactttt ctcgtttagc
     2101 acttagactc acttctgact tttttggtcc aaagtagcag acagcgactg aggaacagga
     2161 agcgggggag cgtatcacat agatgtgttc tgaagacagg gcccatctgc gccccaactt
     2221 gtctacccag cactgcacag agctgctttc atgggtggct gcctcaccca ggctggtcaa
     2281 ctacagtcag gatgctcact gcctcaaggg gtgtgtccaa tctgcccact ccagagtgac
     2341 ggagctcggg ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg
     2401 ggatccagat ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga
     2461 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2521 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     2581 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     2641 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2701 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2761 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2821 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2881 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     2941 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     3001 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa
     3061 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     3121 cgccgggatc actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc
     3181 gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     3241 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3301 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct
     3361 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3421 gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat
     3481 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3541 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3601 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga
     3661 tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3721 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3781 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3841 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3901 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3961 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4021 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4081 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4141 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4201 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4261 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4321 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4381 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4441 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4501 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4561 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4621 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4681 tgtatccgct catgagacaa taaccctgat aaat
//
