LOCUS       sequence-486378-        4591 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  SYFP2-tagged Brain Endothelial Cell  Enhancer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4591)
  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 4591)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 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..4591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      907..916
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(913..915,916..918,919..1629)
                     /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(958..979)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1219..1238)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1566..1587
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1722..1742)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1988..2464
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2187..2206)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2504..2644
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2504..2633
                     /label=AAV2 ITR
     rep_origin      2719..3174
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2806..2825)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3016..3037
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3191..3210)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3310..3332
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3370..3388)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3456..3560
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3561..3629,3630..4421)
                     /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(3779..3798)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtacattg attattgact agtgagctcg ggctgggcat aaaagtcagg
      841 gcagagccat ctattgctta catttgcttc tgggatccag atctttcgaa gctagcgcta
      901 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      961 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1021 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     1081 ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc
     1141 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1201 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1261 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1321 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     1381 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     1441 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1501 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     1561 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1621 ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg
     1681 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     1741 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     1801 ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc
     1861 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggcgcctccc
     1921 gggttcaagc gattctcctg cctcagcctc ccgagggacc gatatgagcg ctgctcgaga
     1981 gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     2041 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     2101 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     2161 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     2221 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     2281 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     2341 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     2401 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     2461 ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     2521 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     2581 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     2641 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2701 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2761 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     2821 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2881 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     2941 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3001 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     3061 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     3121 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     3181 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     3241 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3301 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3361 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     3421 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3481 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3541 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3601 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3661 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3721 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3781 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3841 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3901 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3961 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4021 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4081 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4141 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4201 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4261 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4321 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4381 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4441 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4501 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4561 cagaccccgt agaaaagatc aaaggatctt c
//
