LOCUS       sequence-477969-        6208 bp    DNA     circular SYN 15-DEC-2025
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 6208)
  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 6208)
  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"; 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..6208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     323..344
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             396..452
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             456..476
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             477..1505
                     /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(1595..1615)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1861..2337
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2060..2079)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2377..2517
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2377..2506
                     /label=AAV2 ITR
     rep_origin      2592..3047
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2679..2698)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2889..2910
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3064..3083)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3183..3205
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3243..3261)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3329..3433
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3434..3502,3503..4294)
                     /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(3652..3671)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4465..5053
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4954..4973
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5115..5244
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      5872..5881
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5878..5880,5881..5883,5884..6208,1..386)
                     /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(6184..6203)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
       61 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
      121 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca
      181 acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
      241 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag
      301 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
      361 ctctcggcat ggacgagctg tacaagggat ccggagctac taacttcagc ctgctgaagc
      421 aggctggaga cgtggaggag aaccctggac ccatgcccaa gaagaagagg aaggtgtcca
      481 atttactgac cgtacaccaa aatttgcctg cattaccggt cgatgcaacg agtgatgagg
      541 ttcgcaagaa cctgatggac atgttcaggg atcgccaggc gttttctgag catacctgga
      601 aaatgcttct gtccgtttgc cggtcgtggg cggcatggtg caagttgaat aaccggaaat
      661 ggtttcccgc agaacctgaa gatgttcgcg attatcttct atatcttcag gcgcgcggtc
      721 tggcagtaaa aactatccag caacatttgg gccagctaaa catgcttcat cgtcggtccg
      781 ggctgccacg accaagtgac agcaatgctg tttcactggt tatgcggcgg atccgaaaag
      841 aaaacgttga tgccggtgaa cgtgcaaaac aggctctagc gttcgaacgc actgatttcg
      901 accaggttcg ttcactcatg gaaaatagcg atcgctgcca ggatatacgt aatctggcat
      961 ttctggggat tgcttataac accctgttac gtatagccga aattgccagg atcagggtta
     1021 aagatatctc acgtactgac ggtgggagaa tgttaatcca tattggcaga acgaaaacgc
     1081 tggttagcac cgcaggtgta gagaaggcac ttagcctggg ggtaactaaa ctggtcgagc
     1141 gatggatttc cgtctctggt gtagctgatg atccgaataa ctacctgttt tgccgggtca
     1201 gaaaaaatgg tgttgccgcg ccatctgcca ccagccagct atcaactcgc gccctggaag
     1261 ggatttttga agcaactcat cgattgattt acggcgctaa ggatgactct ggtcagagat
     1321 acctggcctg gtctggacac agtgcccgtg tcggagccgc gcgagatatg gcccgcgctg
     1381 gagtttcaat accggagatc atgcaagctg gtggctggac caatgtaaat attgtcatga
     1441 actatatccg taacctggat agtgaaacag gggcaatggt gcgcctgctg gaagatggcg
     1501 attaagtcga cggcgcgccg cggccgcgaa ttcgatatca taatcaacct ctggattaca
     1561 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     1621 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     1681 ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc
     1741 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggcgcct cccgggttca
     1801 agcgattctc ctgcctcagc ctcccgaatc gggtcggcca gcgctgctcg agagatctac
     1861 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     1921 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     1981 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     2041 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     2101 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     2161 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     2221 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     2281 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     2341 attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     2401 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     2461 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     2521 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     2581 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2641 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     2701 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2761 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     2821 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2881 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     2941 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     3001 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3061 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3121 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3181 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3241 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3301 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3361 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3421 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3481 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3541 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3601 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3661 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3721 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3781 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3841 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3901 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3961 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4021 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4081 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4141 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4201 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4261 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4321 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4381 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4441 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4501 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4561 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4621 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4681 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4741 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4801 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4861 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4921 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4981 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5041 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5101 ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
     5161 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     5221 ccaactccat cactaggggt tcctgcggcc gcacgcgtac attgattatt gactagtagg
     5281 gagcctggaa ttccctggtg aaactgggct ttaaagacaa tgtaggttat cctggtggga
     5341 tggtatacaa ggataacccc agcactggga tgggaggatt accacacatt caaggctaag
     5401 ctggtttcat agtgagattc tgtctccaaa acacacagag gtagctcagt tattagagag
     5461 cttgcccagc ctgtgttaaa ccctgtgtta aagccttggc actccaggag gtggaggcag
     5521 gaagatcaga agttcaaaga catcctcagc tatgcaacaa gggaaacccc acccatctca
     5581 aaagcaaaca ggcaagaaac tgaagtttta ttcctcacat tcctgagact agaaactaaa
     5641 gcacacatgc ctgccctgtg agacaggcat ttatctcctt caaatagatc ctgtgaccct
     5701 cttcccttac acaggctttg cacctgcccc tccttaccca caaggctctg ggacatccac
     5761 ccacagtacc tctaccttga gctcgggctg ggcataaaag tcagggcaga gccatctatt
     5821 gcttacattt gcttctggga tccagatctt tcgaagctag cgctaccggt cgccaccatg
     5881 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     5941 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     6001 aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     6061 gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
     6121 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     6181 aaggacgacg gcaactacaa gacccgcg
//
