LOCUS       sequence-386316-        6044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ratiometric Myr/Palm_mCherry control probe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6044)
  AUTHORS   Czernecki C, Dixit S, Riezman I, Innocenti S, Bornmann C, Pfrieger 
            FW, Riezman H, Scheiffele P
  TITLE     Cell type-specific assessment of cholesterol distribution in models 
            of neurodevelopmental disorders
  JOURNAL   bioRxiv 2022.11.16.516849
REFERENCE   2  (bases 1 to 6044)
  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"; journalName: "bioRxiv 
            2022.11.16.516849"
COMMENT     Sequence Label: pAAV-Syn-Myr_mCherry-2A-eGFP
FEATURES             Location/Qualifiers
     source          1..6044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    96..144
                     /note="synthetic polyadenylation signal"
     primer_bind     198..217
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        219..666
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             699..1400
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELY"
     primer_bind     complement(841..859)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1109..1128
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1404..1457
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(1473..1475,1476..1478,1479..2192)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1473..1475,1476..1478,1479..2189)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1518..1539)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1779..1798)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2126..2147
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2214..2802
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2267..2287)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2685..2696)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2835..2852)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2841..3048
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3057..3186
                     /label=AAV2 ITR
     rep_origin      3292..3745
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3379..3398)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3589..3610
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3812..3831)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3931..3953
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3991..4009)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4077..4181
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4182..4250,4251..5042)
                     /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(4400..4419)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5213..5801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5702..5721
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agagggagtg gccaactcca tcactagggg ttcctcagat ctaggatcac gcgttctaga
       61 aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac
      121 atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca
      181 aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtggg ttttaggacc
      241 aggatgaggc ggggtggggg tgcctacctg acgaccgacc ccgacccact ggacaagcac
      301 ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg aaacaggatg
      361 cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc gcccccgcct
      421 ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact cgccggtccc
      481 ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga
      541 ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg ctgcggcgcc
      601 ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga
      661 gcgcagtcga attcatgggc tgcaccctga gcgcaagcgt gagcaagggc gaggaggata
      721 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
      781 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
      841 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
      901 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
      961 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     1021 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     1081 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     1141 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     1201 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     1261 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     1321 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     1381 gcggcatgga cgagctgtac ctcgagggca gaggaagtct tctaacatgc ggtgacgtgg
     1441 aggagaatcc cggccctgca ccgggatcca ccatggtgag caagggcgag gagctgttca
     1501 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1561 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1621 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1681 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1741 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1801 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1861 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1921 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1981 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2041 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     2101 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     2161 tcactctcgg catggacgag ctgtacaagt aaagcggccg caagcttatc gataatcaac
     2221 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2281 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2341 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     2401 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     2461 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2521 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2581 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     2641 ttgccacctg gattctgcgc gggacgccct tctgctacgt cccttcggcc ctcaatccag
     2701 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     2761 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgactag
     2821 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2881 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2941 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3001 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggaga gatctgagga
     3061 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3121 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     3181 gcgcagagag ggagtggccc cccccccccc ccccctgcag cctggcgtaa tagcgaagag
     3241 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     3301 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3361 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3421 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3481 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3541 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3601 tccaaactgg aacaacactc aaccatctcg gtctattctt ttgatttata agggattttg
     3661 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3721 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     3781 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3841 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3901 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3961 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4021 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4081 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4141 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4201 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4261 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4321 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4381 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4441 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4501 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4561 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4621 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4681 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4741 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4801 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4861 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4921 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4981 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5041 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5101 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5161 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5221 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5281 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5341 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5401 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5461 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5521 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5581 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5641 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5701 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5761 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5821 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5881 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5941 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6001 cgcctctccc cgcgcgttgg ccgattcatt aatgcaggct gcag
//
