LOCUS       sequence-497202-        5092 bp    DNA     circular SYN 13-JAN-2026
DEFINITION  SYFP2-tagged Brain Endothelial Cell Enhancer mini Beta Goblin 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5092)
  AUTHORS   Velazquez-Rivera E, Dey O, Kim NS, Cao W, Ye Q, Gao P, Thai A, 
            Nguyen JK, Zhang H, Ting JT, Gopi M, Ren B, Holmes TC, Xu X
  TITLE     Specific targeting of brain endothelial cells using enhancer AAV 
            vectors.
  JOURNAL   Neuron. 2025 May 21;113(10):1562-1578.e6. doi: 
            10.1016/j.neuron.2025.03.031.
  PUBMED    40403707
REFERENCE   2  (bases 1 to 5092)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 13, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2025.03"; journalName: "Neuron"; date: 
            "2025-05-21- 21"; volume: "113"; issue: "10"; pages: "1562-1578.e6"
COMMENT     Sequence Label: pAAV-BEC enhancer-minBG-SYFP2
FEATURES             Location/Qualifiers
     source          1..5092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(174..762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(254..273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(933..1724,1725..1793))
                     /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     1556..1575
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1794..1898)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1966..1984
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2022..2044)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2144..2163
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2180..2635)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2317..2338)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2529..2548
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2710..2850
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2721..2850
                     /label=AAV2 ITR
     polyA_signal    2890..3366
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3148..3167
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     3612..3632
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3725..4435,4436..4438,4439..4441))
                     /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(3767..3788)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4116..4135
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4375..4396
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4438..4447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   join(5075..5092,1..112)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
       61 cccgacgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggacatgtga
      121 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      181 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      241 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      301 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      361 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      421 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      481 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      541 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      601 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      661 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      721 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      781 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      841 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      901 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
      961 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1021 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1081 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1141 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1201 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1261 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1321 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1381 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1441 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1501 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1561 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1621 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1681 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1741 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1801 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1861 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1921 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     1981 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2041 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2101 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     2161 gtactgagag tgcaccataa aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     2221 ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa
     2281 tcaaaagaat agcccgagat agagttgagt gttgttccag tttggaacaa gagtccacta
     2341 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2401 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     2461 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     2521 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc
     2581 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtactat
     2641 ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2701 tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
     2761 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     2821 gagtggccaa ctccatcact aggggttcct gcggccgctc ggtccgcacg tggttaccta
     2881 caaaatcaga aggacaggga agggagcagt ggttcacgcc tgtaatccca gcaatttggg
     2941 aggccaaggt gggtagatca cctgagatta ggagttggag accagcctgg ccaatatggt
     3001 gaaaccccgt ctctaccaaa aaaacaaaaa ttagctgagc ctggtcatgc atgcctggaa
     3061 tcccaacaac tcgggaggct gaggcaggag aatcgcttga acccaggagg cggagattgc
     3121 agtgagccaa gattgtgcca ctgcactcca gcttggttcc caatagaccc cgcaggccct
     3181 acaggttgtc ttcccaactt gccccttgct ccataccacc cccctccacc ccataatatt
     3241 atagaaggac acctagtcag acaaaatgat gcaacttaat tttattagga caaggctggt
     3301 gggcactgga gtggcaactt ccagggccag gagaggcact ggggaggggt cacagggatg
     3361 ccacccgtag atctctcgag cagcgctggc cgacccgatt cgggaggctg aggcaggaga
     3421 atcgcttgaa cccgggaggc gccacggaat tgtcagtgcc caacagccga gcccctgtcc
     3481 agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aagaactaac caggatttat
     3541 acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag
     3601 cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac
     3661 aaattttgta atccagaggt tgattatgat atcgaattcg cggccgcggc gcgccgtcga
     3721 cttacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca
     3781 ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag cttggactgg tagctcaggt
     3841 agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt
     3901 cggcgagctg cacgccgccg tcctcgatgt tgtggcggat cttgaagttg gccttgatgc
     3961 cgttcttctg cttgtcggcg gtgatataga cgttgtggct gttgtagttg tactccagct
     4021 tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca
     4081 ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga
     4141 agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct
     4201 gcttcatgtg gtcggggtag cgggcgaagc actgcacgcc gtagcccagg gtggtcacga
     4261 gggtgggcca gggcacgggc agcttgccgg tggtgcagat cagcttcagg gtcagcttgc
     4321 cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc
     4381 cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca
     4441 tggtggcgac cggtagcgct agcttcgaaa gatctggatc ccagaagcaa atgtaagcaa
     4501 tagatggctc tgccctgact tttatgccca gcccgagctc aaggtagagg tactgtgggt
     4561 ggatgtccca gagccttgtg ggtaaggagg ggcaggtgca aagcctgtgt aagggaagag
     4621 ggtcacagga tctatttgaa ggagataaat gcctgtctca cagggcaggc atgtgtgctt
     4681 tagtttctag tctcaggaat gtgaggaata aaacttcagt ttcttgcctg tttgcttttg
     4741 agatgggtgg ggtttccctt gttgcatagc tgaggatgtc tttgaacttc tgatcttcct
     4801 gcctccacct cctggagtgc caaggcttta acacagggtt taacacaggc tgggcaagct
     4861 ctctaataac tgagctacct ctgtgtgttt tggagacaga atctcactat gaaaccagct
     4921 tagccttgaa tgtgtggtaa tcctcccatc ccagtgctgg ggttatcctt gtataccatc
     4981 ccaccaggat aacctacatt gtctttaaag cccagtttca ccagggaatt ccaggctccc
     5041 tactagtcaa taatcaatgt acgcgtgcgg ccgcaggaac ccctagtgat gg
//
