LOCUS       sequence-502701-        6267 bp    DNA     circular SYN 26-MAR-2026
DEFINITION  Neuronal overexpression of synaptophysin fused to mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6267)
  AUTHORS   Ramirez JJ, Hardin EJ, Sakers K, Kim J, Colon Ortiz C, Savage JT, 
            Hanamsagar R, Block CL, Singh SK, Bilbo SD, Eroglu C
  TITLE     Astrocyte-microglia crosstalk through Hevin and Toll-like receptor 
            signaling controls developmental thalamocortical synapse refinement.
  JOURNAL   Neuron. 2026 Feb 19:S0896-6273(25)00984-5. doi: 
            10.1016/j.neuron.2025.12.028.
  PUBMED    41720089
REFERENCE   2  (bases 1 to 6267)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 26, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Neuron. 
            2026 Feb 19:S0896-6273(25)00984-5. doi: 
            10.1016/j.neuron.2025.12.028."
COMMENT     Sequence Label: pAAV-hsyn1-synaptophysin-mCherry-V5
FEATURES             Location/Qualifiers
     source          1..6267
                     /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"
     promoter        825..1272
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1292..1301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2267..2974
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2412..2430)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2680..2699
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2874..2897
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2975..3016
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3044..3632
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3097..3117)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3515..3526)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3664..4140
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3863..3882)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4180..4320
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4180..4309
                     /label=AAV2 ITR
     rep_origin      4395..4850
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4482..4501)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4692..4713
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4867..4886)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4986..5008
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5046..5064)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5132..5236
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5237..5305,5306..6097)
                     /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(5455..5474)
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca agtgggtttt
      841 aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac
      901 aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac
      961 aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc
     1021 ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc
     1081 ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca
     1141 gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc
     1201 ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc
     1261 ctgagagcgc agtcgagaag gtaccggatc cgccaccatg gacgtggtga atcagctggt
     1321 ggctgggggt cagttccggg tggtcaagga gccccttggc ttcgtgaagg tgctgcagtg
     1381 ggtctttgcc atcttcgcct ttgctacgtg tggcagctac accggggagc ttcggctgag
     1441 cgtggagtgt gccaacaaga cggagagtgc cctcaacatc gaagttgaat tcgagtaccc
     1501 cttcaggctg caccaagtgt actttgatgc accctcctgc gtcaaagggg gcactaccaa
     1561 gatcttcctg gttggggact actcctcgtc ggctgaattc tttgtcaccg tggctgtgtt
     1621 tgccttcctc tactccatgg gggccctggc cacctacatc ttcctgcaga acaagtaccg
     1681 agagaacaac aaagggccta tgatggactt tctggctaca gccgtgttcg ctttcatgtg
     1741 gctagttagt tcatcagcct gggccaaagg cctgtccgat gtgaagatgg ccacggaccc
     1801 agagaacatt atcaaggaga tgcccatgtg ccgccagaca gggaacacat gcaaggaact
     1861 gagggaccct gtgacttcag gactcaacac ctcagtggtg tttggcttcc tgaacctggt
     1921 gctctgggtt ggcaacttat ggttcgtgtt caaggagaca ggctgggcag ccccattcat
     1981 gcgcgcacct ccaggcgccc cggaaaagca accagcacct ggcgatgcct acggcgatgc
     2041 gggctacggg cagggccccg gaggctatgg gccccaggac tcctacgggc ctcagggtgg
     2101 ttatcaaccc gattacgggc agccagccag cggtggcggt ggctacgggc ctcagggcga
     2161 ctatgggcag caaggctatg gccaacaggg tgcgcccacc tccttctcca atcagatggg
     2221 atccactagt aacggccgcc agtgtgctgg aattctgcag atatcgatgg tgagcaaggg
     2281 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     2341 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     2401 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
     2461 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     2521 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     2581 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     2641 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     2701 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     2761 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgccgagg tcaagaccac
     2821 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagctgga
     2881 catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgcg ccgagggccg
     2941 ccactccacc ggcggcatgg acgagctgta caagggtaag cctatcccta accctctcct
     3001 cggtctcgat tctacgtagg aattcgatat caagcttatc gataatcaac ctctggatta
     3061 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3121 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3181 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3241 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3301 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3361 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3421 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg ttgccacctg
     3481 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3541 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3601 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc
     3661 tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact
     3721 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     3781 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     3841 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct
     3901 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag
     3961 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     4021 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     4081 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     4141 ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     4201 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4261 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     4321 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     4381 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4441 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     4501 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4561 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4621 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4681 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     4741 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     4801 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     4861 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4921 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4981 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     5041 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     5101 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5161 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5221 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5281 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5341 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5401 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5461 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5521 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5581 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5641 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5701 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5761 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5821 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5881 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5941 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6001 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6061 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6121 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6181 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6241 ccccgtagaa aagatcaaag gatcttc
//
