LOCUS       sequence-468722-        6441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expression of eGFP and wild-type Tau.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6441)
  TITLE     Nolan lab unpublished
REFERENCE   2  (bases 1 to 6441)
  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"
COMMENT     Sequence Label: pAAV FLEX eGFP 2A Tau WT WPRE
FEATURES             Location/Qualifiers
     source          1..6441
                     /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"
     primer_bind     complement(830..850)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        835..1164
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1015..1150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1374..1598
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1587..1604
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    1631..2219
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1737..1748
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2146..2166
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    2258..2291
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3220..3253)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3316..3349)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(3384..3403)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3465..3487)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3766..4044)
                     /label=chicken beta-actin promoter
     repeat_region   4350..4479
                     /label=AAV2 ITR
     promoter        complement(4500..4518)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4501..4518)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4808..4827)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5041..5060)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5160..5182
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5220..5238)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5306..5410
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5411..5479,5480..6271)
                     /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(5629..5648)
                     /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 tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgtg gaatgtgtgt
      841 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
      901 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      961 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     1021 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1081 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     1141 tttggaggcc taggcttttg caaaaagctc tgcgcgctcg ctcgctcact gaggccgccc
     1201 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
     1261 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
     1321 tctacgtagc catgctctag tcgagcccca gctggttctt tccgcctcag aagccataga
     1381 gcccaccgca tccccagcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg
     1441 ccccacccca ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc
     1501 attttattag gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga
     1561 ggggcaaaca acagatggct ggcaactaga aggcacagtc gaggctgatc agcgagtcga
     1621 cggtatcgat gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga
     1681 agacgcggaa gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg
     1741 agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacacaggcg
     1801 agcagccaag gaaaggacga tgatttcccc gacaacacca cggaattgtc agtgcccaac
     1861 agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata
     1921 gggaggggga aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca
     1981 gtgggggttg cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac
     2041 aactcctcat aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata
     2101 cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga
     2161 gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta
     2221 tcgataagct gttaaagctt gaacctcttc gagggacata acttcgtata atgtatgcta
     2281 tacgaagtta tgggtcgatg gtgagatcgc cacaccgcgg ggagcagccc ctccaggcca
     2341 gaagggccag gccaacgcca ccaggattcc agcaaaaacc ccgcccgctc caaagacacc
     2401 acccagctct ggtgaacctc caaaatcagg ggatcgcagc ggctacagca gccccggctc
     2461 cccaggcact cccggcagcc gctcccgcac cccgtccctt ccaaccccac ccacccggga
     2521 gcccaagaag gtggcagtgg tccgtactcc acccaagtcg ccgtcttccg ccaagagccg
     2581 cctgcagaca gcccccgtgc ccatgccaga cctgaagaat gtcaagtcca agatcggctc
     2641 cactgagaac ctgaagcacc agccgggagg cgggaaggtg cagataatta ataagaagct
     2701 ggatcttagc aacgtccagt ccaagtgtgg ctcaaaggat aatatcaaac acgtcccggg
     2761 aggcggcagt gtgcaaatag tctacaaacc agttgacctg agcaaggtga cctccaagtg
     2821 tggctcatta ggcaacatcc atcataaacc aggaggtggc caggtggaag taaaatctga
     2881 gaagcttgac ttcaaggaca gagtccagtc gaagattggg tccctggaca atatcaccca
     2941 cgtccctggc ggaggaaata aaaagattga aacccacaag ctgaccttcc gcgagaacgc
     3001 caaagccaag acagaccacg gggcggagat cgtgtacaag tcgccagtgg tgtctgggga
     3061 cacgtctcca cggcatctca gcaatgtctc ctccaccggc agcatcgaca tggtagactc
     3121 gccccagctc gccacgctag ctgacgaggt gtctgcctcc ctggccaagc agggtttgta
     3181 gactagcgcg gccgcactag tgagctcctg caggaattca taacttcgta tagcatacat
     3241 tatacgaagt tatgggtcga tggtgatgct tggcaattcg ggtcgatggt gaagcattgg
     3301 tcttcctatt ttgatataac ttcgtatagg atactttata cgaagttatg gggatccaat
     3361 tctttgccaa aatgatgaga cagcacaata accagcacgt tgcccaggag ctgtaggaaa
     3421 aagaagaagg catgaacatg gttagcagag gctctagagc cgccggtcac acgccagaag
     3481 ccgaaccccg ccctgccccg tcccccccga aggcagccgt ccccccgcgg acagccccga
     3541 gccgctcccc ccgcacaaag ggccctcccg gagcccctca aggctttcac gcagccacag
     3601 aaaagaaaca agccgtcatt aaaccaagcg ctaattacag cccggaggag aagggccgtc
     3661 ccgcccgctc acctgtggga gtaacgcggt cagtcagagc cggggcgggc ggcgcgaggc
     3721 ggcggcggag cggggcacgg ggcgaaggca gcgcgcagcg actcccgccc gccgcgcgct
     3781 tcgcttttta tagggccgcc gccgccgccg cctcgccata aaaggaaact ttcggagcgc
     3841 gccgctctga ttggctgccg ccgcacctct ccgcctcgcc ccgccccgcc cctcgccccg
     3901 ccccgccccg cctggcgcgc gccccccccc ccccccccgc ccccatcgct gcacaaaata
     3961 attaaaaaat aaataaatac aaaattgggg gtggggaggg gggggagatg gggagagtga
     4021 agcagaacgt ggggctcacc tcgaccatgg taatagcgat gactaatacg tagatgtact
     4081 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     4141 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     4201 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     4261 ctatgggaac atacgtcatt attgacgtca atggtaccta gagcatggct acgtagataa
     4321 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     4381 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     4441 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tagaattaat tccgtgtatt
     4501 ctatagtgtc acctaaatcg tatgtgtatg atacataagg ttatgtatta attgtagccg
     4561 cgttctaacg acaatatgta caagcctaat tgtgtagcat ctggcttact gaagcagacc
     4621 ctatcatctc tctcgtaaac tgccgtcaga gtcggtttgg ttggacgaac cttctgagtt
     4681 tctggtaacg ccgtcccgca cccggaaatg gtcagcgaac caatcagcag ggtcatcgct
     4741 agccagatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
     4801 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     4861 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
     4921 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
     4981 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgaatggt gcactctcag
     5041 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5101 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5161 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     5221 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     5281 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5341 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5401 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5461 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5521 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5581 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5641 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5701 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5761 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5821 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5881 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5941 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6001 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6061 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6121 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6181 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6241 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6301 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6361 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6421 agaaaagatc aaaggatctt c
//
