LOCUS       sequence-470918-        5119 bp    DNA     circular SYN 30-APR-2025
DEFINITION  AiE0254h 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 5119)
  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 5119)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 30, 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: AiP11772 -
            pAAV-hsA2-AiE0254h-minRho-SYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..5119
                     /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      1156..1165
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1162..1164,1165..1167,1168..1878)
                     /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(1207..1228)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1468..1487)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1815..1836
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1946..1966)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2153..2264
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2382..2522
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2382..2511
                     /label=AAV2 ITR
     rep_origin      2597..3052
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2684..2703)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2894..2915
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3069..3088)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3188..3210
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3248..3266)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3334..3438
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3439..3507,3508..4299)
                     /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(3657..3676)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4470..5058
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4959..4978
                     /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 gcggccgcaa cgcgtttaga acaatggctg gcccatagta aatgccgtgt
      181 tagtgtgtta gttgctgttc ttccacgtca gaagaggcac agacaaatta ccaccaggtg
      241 gcgctcagag tctgcggagg catcacaaca gccctgaatt tgaatcctgc tctgccactg
      301 cctagttgag accttttact acctgactag ctgtttgtgt attttaggtg tttgtttctt
      361 gcacgcttat tcaggctttt ggcagaaatc agttcgttgt ggtttaagga ctgaggtcgc
      421 ctttctttgc tgctaactgt tggccaggaa gtactgtcag ctactacatg gtacttgctg
      481 gtccttgcac atgaccccct ccaccttcaa accaggattg acacagtgtg ttgagtcttt
      541 cttgaagact tttgaacatc tgacttccca ttctgccact agccagttaa aattctctaa
      601 ttttgaagga ctcacctggt tggccaaacc cacccaaata atctccacat cttgaagtca
      661 actgacccag gactttacat atgcaaaatc ccttcacagc agtacctaga tgagtgtttg
      721 cttgaataac tgggacacag gaatcttggg ggagccatct ttagaattcg acctcctaca
      781 acccttctgg aaatctgaga gtgagtcagg ggaagaaacc ctcttttgta gtttcctttt
      841 agggctttct actttgctca aagttgggca ctatttcact tcagtagggt cctgcaagcc
      901 ccatgagggt agtgagtgct gtcctaggaa acagtaactt aaccctgata cccatttgtc
      961 caagaattcg atatcataat caaccatagg taccgagctc gggattcagc cgggagctta
     1021 gggaggggag gtcacttcat aagggcctgg ggggggagtt ggagccacga gtcgtccagc
     1081 cggagccccg tgtggctgag ctccggcctc agaagcatcc ccgggttgga tccttcgaag
     1141 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1201 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1261 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
     1321 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcgtgca gtgcttcgcc
     1381 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1441 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1501 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1561 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1621 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     1681 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1741 gtgctgctgc ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac
     1801 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1861 atggacgagc tgtacaagta agtcgacatc ataatcaacc tctggattac aaaatttgtg
     1921 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1981 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     2041 aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2101 caggggctcg gctgttgggc actgacaatt ccgtggctcg agagatcttc gactgtgcct
     2161 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2221 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2281 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2341 aatagcaggc atgagatctc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     2401 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     2461 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     2521 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     2581 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2641 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     2701 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2761 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     2821 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2881 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     2941 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     3001 atttaacaaa aatttaacgg gaattttaac aaaatattaa cgtttacaat tttatggtgc
     3061 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3121 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3181 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3241 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3301 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3361 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3421 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3481 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3541 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3601 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3661 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3721 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3781 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3841 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3901 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3961 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4021 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4081 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4141 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4201 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4261 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4321 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4381 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4441 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4501 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4561 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4621 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4681 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4741 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4801 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4861 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4921 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4981 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5041 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5101 tggccttttg ctcacatgt
//
