LOCUS       sequence-477968-        5425 bp    DNA     circular SYN 09-JUL-2025
DEFINITION  Cre Brain Endothelial Cell Enhancer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5425)
  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 5425)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 9, 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-Cre
FEATURES             Location/Qualifiers
     source          1..5425
                     /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"
     CDS             1417..1437
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1438..2466
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     primer_bind     complement(2556..2576)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2822..3298
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3021..3040)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3338..3478
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3338..3467
                     /label=AAV2 ITR
     rep_origin      3553..4008
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3640..3659)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3850..3871
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4025..4044)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4144..4166
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4204..4222)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4290..4394
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4395..4463,4464..5255)
                     /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(4613..4632)
                     /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 agtagggagc ctggaattcc ctggtgaaac
      841 tgggctttaa agacaatgta ggttatcctg gtgggatggt atacaaggat aaccccagca
      901 ctgggatggg aggattacca cacattcaag gctaagctgg tttcatagtg agattctgtc
      961 tccaaaacac acagaggtag ctcagttatt agagagcttg cccagcctgt gttaaaccct
     1021 gtgttaaagc cttggcactc caggaggtgg aggcaggaag atcagaagtt caaagacatc
     1081 ctcagctatg caacaaggga aaccccaccc atctcaaaag caaacaggca agaaactgaa
     1141 gttttattcc tcacattcct gagactagaa actaaagcac acatgcctgc cctgtgagac
     1201 aggcatttat ctccttcaaa tagatcctgt gaccctcttc ccttacacag gctttgcacc
     1261 tgcccctcct tacccacaag gctctgggac atccacccac agtacctcta ccttgagctc
     1321 gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca
     1381 gatctttcga agctagcgct accggtcgcc accatgccca agaagaagag gaaggtgtcc
     1441 aatttactga ccgtacacca aaatttgcct gcattaccgg tcgatgcaac gagtgatgag
     1501 gttcgcaaga acctgatgga catgttcagg gatcgccagg cgttttctga gcatacctgg
     1561 aaaatgcttc tgtccgtttg ccggtcgtgg gcggcatggt gcaagttgaa taaccggaaa
     1621 tggtttcccg cagaacctga agatgttcgc gattatcttc tatatcttca ggcgcgcggt
     1681 ctggcagtaa aaactatcca gcaacatttg ggccagctaa acatgcttca tcgtcggtcc
     1741 gggctgccac gaccaagtga cagcaatgct gtttcactgg ttatgcggcg gatccgaaaa
     1801 gaaaacgttg atgccggtga acgtgcaaaa caggctctag cgttcgaacg cactgatttc
     1861 gaccaggttc gttcactcat ggaaaatagc gatcgctgcc aggatatacg taatctggca
     1921 tttctgggga ttgcttataa caccctgtta cgtatagccg aaattgccag gatcagggtt
     1981 aaagatatct cacgtactga cggtgggaga atgttaatcc atattggcag aacgaaaacg
     2041 ctggttagca ccgcaggtgt agagaaggca cttagcctgg gggtaactaa actggtcgag
     2101 cgatggattt ccgtctctgg tgtagctgat gatccgaata actacctgtt ttgccgggtc
     2161 agaaaaaatg gtgttgccgc gccatctgcc accagccagc tatcaactcg cgccctggaa
     2221 gggatttttg aagcaactca tcgattgatt tacggcgcta aggatgactc tggtcagaga
     2281 tacctggcct ggtctggaca cagtgcccgt gtcggagccg cgcgagatat ggcccgcgct
     2341 ggagtttcaa taccggagat catgcaagct ggtggctgga ccaatgtaaa tattgtcatg
     2401 aactatatcc gtaacctgga tagtgaaaca ggggcaatgg tgcgcctgct ggaagatggc
     2461 gattaagtcg acggcgcgcc gcggccgcga attcgatatc ataatcaacc tctggattac
     2521 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     2581 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2641 tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc
     2701 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggcgcc tcccgggttc
     2761 aagcgattct cctgcctcag cctcccgaat cgggtcggcc agcgctgctc gagagatcta
     2821 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     2881 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     2941 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     3001 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     3061 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     3121 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     3181 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     3241 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     3301 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     3361 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3421 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     3481 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     3541 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3601 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     3661 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3721 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     3781 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3841 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     3901 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     3961 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     4021 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4081 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4141 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     4201 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4261 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4321 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4381 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4441 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4501 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4561 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4621 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4681 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4741 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4801 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4861 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4921 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4981 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5041 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5101 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5161 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5221 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5281 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5341 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5401 ccgtagaaaa gatcaaagga tcttc
//
