LOCUS       sequence-410482-        4458 bp    DNA     circular SYN 17-APR-2025
DEFINITION  Direct-expressing SYFP2 in astrocytes AAV Virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4458)
  AUTHORS   Mich JK, Sunil S, Johansen N, Martinez RA, Leytze M, Gore BB, 
            Mahoney JT, Ben-Simon Y, Bishaw Y, Brouner K, Campos J, Canfield R, 
            Casper T, Dee N, Egdorf T, Gary A, Gibson S, Goldy J, Groce EL, 
            Hirschstein D, Loftus L, Lusk N, Malone J, Martin NX, Monet D, 
            Omstead V, Opitz-Araya X, Oster A, Pom CA, Potekhina L, Reding M, 
            Rimorin C, Ruiz A, Sedeno-Cortes AE, Shapovalova NV, Taormina M, 
            Taskin N, Tieu M, Valera Cuevas NJ, Weed N, Way S, Yao Z, McMillen 
            DA, Kunst M, McGraw M, Thyagarajan B, Waters J, Bakken TE, Yao S, 
            Smith KA, Svoboda K, Podgorski K, Kojima Y, Horwitz GD, Zeng H, 
            Daigle TL, Lein ES, Tasic B, Ting JT, Levi BP
  TITLE     Enhancer-AAVs allow genetic access to oligodendrocytes and diverse 
            populations of astrocytes across species
  JOURNAL   bioRxiv 2023.09.20.558718
REFERENCE   2  (bases 1 to 4458)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2023.09.20.558718"
COMMENT     Sequence Label: AiP12146 -
            pAAV-AiE0381h-minBglobin-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4458
                     /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      470..479
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(476..478,479..481,482..1192)
                     /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(521..542)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(782..801)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1129..1150
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1285..1305)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1492..1603
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1721..1861
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1721..1850
                     /label=AAV2 ITR
     rep_origin      1936..2391
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2023..2042)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2233..2254
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2408..2427)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2527..2549
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2587..2605)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2673..2777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2778..2846,2847..3638)
                     /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(2996..3015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3809..4397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4298..4317
                     /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 gcggccgcac gcgtgggctt tctaaactgg cctggaacac tgttagttaa
      181 taaacattcc tttcagggtt gttctctttg cttctgatct cattgatgat gatttgttgg
      241 acaaataagg aagtaaacag tgcagagttt tttggcttct gctctattaa aaacatagca
      301 gagaaggaca agaagttcct gcctgctaaa taggaagatc tagggaatca aaagagtctc
      361 tgtcaagctt cttgtagagc tcgggctggg cataaaagtc agggcagagc catctattgc
      421 ttacatttgc ttctgggatc cagatctttc gaagctagcg ctaccggtcg ccaccatggt
      481 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
      541 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
      601 gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
      661 gaccaccctg ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
      721 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
      781 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
      841 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
      901 ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat
      961 caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca
     1021 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1081 gagctaccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1141 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaagtcga
     1201 cggcgcgccg cggccgcgaa ttcgatatca taatcaacct ctggattaca aaatttgtga
     1261 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     1321 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     1381 atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     1441 aggggctcgg ctgttgggca ctgacaattc cgtggctcga gagatcttcg actgtgcctt
     1501 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1561 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     1621 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     1681 atagcaggca tgagatctca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     1741 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     1801 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     1861 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     1921 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1981 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     2041 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2101 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     2161 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2221 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     2281 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     2341 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     2401 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     2461 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     2521 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     2581 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2641 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2701 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2761 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2821 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2881 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2941 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3001 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3061 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3121 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3181 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3241 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3301 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3361 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3421 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3481 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3541 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3601 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3661 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3721 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3781 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3841 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3901 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3961 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4021 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4081 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4141 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4201 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4261 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4321 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4381 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4441 ggccttttgc tcacatgt
//
