LOCUS       sequence-480280-        5640 bp    DNA     circular SYN 24-JUL-2025
DEFINITION  Brain Endothelial Cell Enhancer with 4x6T miRNA targeting cassette 
            For use in combination with Cre-dependent transgenic mouse lines or 
            co-infection with Cre dependent viral vectors..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5640)
  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 5640)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 24, 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-iCre(R279T)-4x6T
FEATURES             Location/Qualifiers
     source          1..5640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..131
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      747..756
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             759..779
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2388..2864
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2587..2606)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2904..3044
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2904..3033
                     /label=AAV2 ITR
     rep_origin      3119..3574
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3206..3225)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3416..3437
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3591..3610)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3710..3732
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3770..3788)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3856..3960
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3961..4029,4030..4821)
                     /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(4179..4198)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4992..5580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5481..5500
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc tgcggccgca cgcgtacatt gattattgac tagtagggag cctggaattc
      181 cctggtgaaa ctgggcttta aagacaatgt aggttatcct ggtgggatgg tatacaagga
      241 taaccccagc actgggatgg gaggattacc acacattcaa ggctaagctg gtttcatagt
      301 gagattctgt ctccaaaaca cacagaggta gctcagttat tagagagctt gcccagcctg
      361 tgttaaaccc tgtgttaaag ccttggcact ccaggaggtg gaggcaggaa gatcagaagt
      421 tcaaagacat cctcagctat gcaacaaggg aaaccccacc catctcaaaa gcaaacaggc
      481 aagaaactga agttttattc ctcacattcc tgagactaga aactaaagca cacatgcctg
      541 ccctgtgaga caggcattta tctccttcaa atagatcctg tgaccctctt cccttacaca
      601 ggctttgcac ctgcccctcc ttacccacaa ggctctggga catccaccca cagtacctct
      661 accttgagct cgggctgggc ataaaagtca gggcagagcc atctattgct tacatttgct
      721 tctgggatcc agatctttcg aagctagcca ccatggtgcc caagaagaag aggaaagtct
      781 ccaacctgct gactgtgcac caaaacctgc ctgccctccc tgtggatgcc acctctgatg
      841 aagtcaggaa gaacctgatg gacatgttca gggacaggca ggccttctct gaacacacct
      901 ggaagatgct cctgtctgtg tgcagatcct gggctgcctg gtgcaagctg aacaacagga
      961 aatggttccc tgctgaacct gaggatgtga gggactacct cctgtacctg caagccagag
     1021 gcctggctgt gaagaccatc caacagcacc tgggccagct caacatgctg cacaggagat
     1081 ctggcctgcc tcgcccttct gactccaatg ctgtgtccct ggtgatgagg agaatcagaa
     1141 aggagaatgt ggatgctggg gagagagcca agcaggccct ggcctttgaa cgcactgact
     1201 ttgaccaagt cagatccctg atggagaact ctgacagatg ccaggacatc aggaacctgg
     1261 ccttcctggg cattgcctac aacaccctgc tgcgcattgc cgaaattgcc agaatcagag
     1321 tgaaggacat ctcccgcacc gatggtggga gaatgctgat ccacattggc aggaccaaga
     1381 ccctggtgtc cacagctggt gtggagaagg ccctgtccct gggggttacc aagctggtgg
     1441 agagatggat ctctgtgtct ggtgtggctg atgaccccaa caactacctg ttctgccggg
     1501 tcagaaagaa tggtgtggct gccccttctg ccacctccca actgtccacc cgggccctgg
     1561 aagggatctt tgaggccacc caccgcctga tctatggtgc caaggatgac tctgggcaga
     1621 gatacctggc ctggtctggc cactctgcca gagtgggtgc tgccaccgac atggccaggg
     1681 ctggtgtgtc catccctgaa atcatgcagg ctggtggctg gaccaatgtg aacattgtga
     1741 tgaactacat cagaaacctg gactctgaga ctggggccat ggtgaggctg ctcgaagatg
     1801 gggactgagg cgcgccgtga tgacatatgg gcattcaccg cgtgccttac tacgcgtatt
     1861 cttaagcaat aagcgaatat aacacggtcg atctaaaaag gttagctggg tgtgttagaa
     1921 gccctgcaag acgacctgcg cattattact cacggtacga cgcattatta ctcacggtac
     1981 gaggcattca ccgcgtgcct tactacgcgt attcttaagc aataagcgaa tataacacgg
     2041 tcgatctaaa aaggttagct gggtgtgtta gaagccctgc aagacgacct gggcattcac
     2101 cgcgtgcctt actacgcgta ttcttaagca ataagcgaat ataacacggt cgatctaaaa
     2161 aggttagctg ggtgtgttag aagccctgca agacgacctg ggtaccgcat tattactcac
     2221 ggtacgacgc attattactc acggtacgag gcattcaccg cgtgccttac tacgcgtatt
     2281 cttaagcaat aagcgaatat aacacggtcg atctaaaaag gttagctggg tgtgttagaa
     2341 gccctgcaag acgacctggg taccaggtaa ccgctcgaga gatctacggg tggcatccct
     2401 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     2461 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     2521 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     2581 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     2641 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     2701 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     2761 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     2821 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa
     2881 ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     2941 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     3001 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     3061 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     3121 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3181 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3241 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3301 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     3361 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3421 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     3481 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3541 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     3601 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     3661 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3721 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     3781 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     3841 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3901 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3961 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4021 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4081 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4141 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4201 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4261 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4321 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4381 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4441 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4501 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4561 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4621 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4681 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4741 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4801 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4861 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4921 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4981 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5041 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5101 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5161 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5221 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5281 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5341 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5401 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5461 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5521 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5581 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
//
