LOCUS       sequence-325691-        5167 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing H2BSYFP2 AAV Virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5167)
  AUTHORS   Graybuck LT, Daigle TL, Sedeno-Cortes AE, Walker M, Kalmbach B, Lenz
            GH, Morin E, Nguyen TN, Garren E, Bendrick JL, Kim TK, Zhou T, 
            Mortrud M, Yao S, Siverts LA, Larsen R, Gore BB, Szelenyi ER, Trader
            C, Balaram P, van Velthoven CTJ, Chiang M, Mich JK, Dee N, Goldy J, 
            Cetin AH, Smith K, Way SW, Esposito L, Yao Z, Gradinaru V, Sunkin 
            SM, Lein E, Levi BP, Ting JT, Zeng H, Tasic B
  TITLE     Enhancer viruses for combinatorial cell-subclass-specific labeling.
  JOURNAL   Neuron. 2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011.
  PUBMED    33789083
REFERENCE   2  (bases 1 to 5167)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Neuron. 
            2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011."
COMMENT     Sequence Label: AiP11496 -
            pAAV-hDlxI56i-minBG-H2B-nsSYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..5167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             787..1164
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             join(1210..1212,1213..1215,1216..1926)
                     /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(1255..1276)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1516..1535)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1863..1884
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1994..2014)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2201..2312
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2430..2570
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2430..2559
                     /label=AAV2 ITR
     rep_origin      2645..3100
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2732..2751)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2942..2963
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3117..3136)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3236..3258
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3296..3314)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3382..3486
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3487..3555,3556..4347)
                     /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(3705..3724)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4518..5106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5007..5026
                     /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 gcgtataggt accgagctct atgcactcac agtggtttgg
      181 catgcatctg gtgaattttt tttaacgaaa aattagtgtt ggtttcgatg tatggtagca
      241 ttctccctaa cgtaatttga ataattcagc aaagccccac taccagctgt acttctgcag
      301 cctcttccat tcttttcagc attataattt tggttaattt tcaattttag gtcctacgtc
      361 tctgcaattt gtgtatgaat aacagaataa tttccctctt ttgtttcgcc tttcctgttc
      421 ctgaatctaa ataaagatgg ctttttagta ttaaaagtgg aagaaaatta caggtaatta
      481 tctttgacgg taaaaacgct gtaatcagcg ggctacatga aaaattactc taattatggc
      541 tgcatttaag agaatggaaa aaaaccttct tgtggataaa aaccttaaat tgtccccaat
      601 gtctgcttca aattggatgg cactgcagct ggaggctttg ttcagaattg atcctgggga
      661 gctacgaacc caaagtttca cagtagggag ctcgggctgg gcataaaagt cagggcagag
      721 ccatctattg cttacatttg cttctgggat ccagatcttt cgaagctagc gctaccggtc
      781 gccaccatgc cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg
      841 gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat
      901 tccatctatg tgtacaaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag
      961 gccatgggca tcatgaattc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct
     1021 tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc
     1081 gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc
     1141 atcaccaagt acaccagcgc taagggaggg ggggggtctg gtggcggggg atctggagga
     1201 ggtggttcca tggtcagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1261 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1321 gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc
     1381 tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg ctaccccgac
     1441 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1501 accatcttct tcaaagacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1561 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1621 ctggggcaca agctggagta caactacaac agccacaacg tctatatcac cgccgacaag
     1681 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     1741 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1801 gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     1861 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1921 tacaaataag tcgacatcat aatcaacctc tggattacaa aatttgtgaa agattgactg
     1981 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2041 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag
     2101 ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     2161 tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag
     2221 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2281 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2341 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2401 gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     2461 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     2521 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     2581 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     2641 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2701 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     2761 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2821 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     2881 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2941 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     3001 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     3061 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     3121 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     3181 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     3241 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     3301 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     3361 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3421 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3481 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3541 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3601 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3661 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3721 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3781 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3841 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3901 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3961 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4021 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4081 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4141 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4201 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4261 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4321 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4381 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4441 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4501 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4561 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4621 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4681 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4741 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4801 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4861 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4921 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4981 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5041 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5101 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5161 cacatgt
//
