LOCUS       hSyn-PreSynBar-h        7462 bp DNA     circular SYN 01-OCT-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7462)
  AUTHORS   Stanford University
  TITLE     Direct Submission
  JOURNAL   Exported Oct 1, 2025 from SnapGene 7.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..130
                     /label=ITR
                     /label=ITR(1)
                     /note="color: #00ffff"
     misc_feature    151..628
                     /label=synapsin promoter
                     /note="color: #4eff51"
     misc_feature    642..646
                     /label=Kozak consensus
                     /note="color: #ffd92b"
     CDS             648..692
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /note="color: #993366"
                     /translation="MSALLILALVGAAVA"
     misc_feature    693..734
                     /label=V5 tag
                     /note="color: #ff80ff"
     CDS             747..2516
                     /codon_start=1
                     /label=Nrxn1b-splitGFP-ORF
                     /note="color: #993366"
                     /translation="ASSLGAHHIHHFHGSSKHHSVPIAIYRSPASLRGGHAGTTYIFSK
                     GGGQITYKWPPNDRPSTRADRLAIGFSTVQKEAVLVRVDSSSGLGDYLELHIHQGKIGV
                     KFNVGTDDIAIEESNAIINDGKYHVVRFTRSGGNATLQVDSWPVIERYPASGGGSGGGS
                     GGGMGGTSMSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLKFICTTG
                     KLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGKYKTRA
                     VVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFTVRH
                     NVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQTVLSKDPNEKGGGSGGGSGGSGGR
                     QLTIFNSQATIIIGGKEQGQPFQGQLSGLYYNGLKVLNMAAENDANIAIVGNVRLVGEV
                     PSSMTTESTATAMQSEMSTSIMETTTTLATSTARRGKPPTKEPISQTTDDILVASAECP
                     SDDEDIDPCEPSSGGLANPTRVGGREPYPGSAEVIRESSSTTGMVVGIVAAAALCILIL
                     LYAMYKYRNRDEGS"
     misc_feature    1209..1937
                     /label=GFP/1-10/+/L
                     /note="color: #a6acb3"
     CDS             2514..2516
                     /codon_start=1
                     /label=insertion site
                     /note="color: #993366"
                     /translation="S"
     misc_feature    2517..2585
                     /label=linker1
                     /note="color: #a6acb3"
     regulatory      2580..2589
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     misc_feature    2586..2651
                     /label=lambda peptide
                     /note="color: #a6acb3"
     misc_feature    2652..2731
                     /label=linker2
                     /note="color: #a6acb3"
     CDS             2739..2870
                     /codon_start=1
                     /label=Nrxn1b-splitGFP-ORF
                     /note="color: #993366"
                     /translation="VDESRNYISNSAQSNGAVVKEKQPSSAKSANKNKKNKDKEYYV"
     misc_feature    2880..3488
                     /label=WPRE
                     /note="color: #a0ff99"
     misc_feature    3502..3726
                     /label=PolyA
                     /note="color: #ff2549"
     misc_feature    3734..4211
                     /label=synapsin promoter
                     /note="color: #4eff51"
     misc_feature    4224..4398
                     /label=PPIB
                     /note="color: #ff0000"
     misc_feature    4411..4431
                     /label=RPL7-5'
                     /note="color: #a6acb3"
     misc_feature    4432..4542
                     /label=2BoxB
                     /note="color: #a6acb3"
     protein_bind    4479..4497
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
                     /note="color: #31849b"
     protein_bind    4503..4521
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
                     /note="color: #31849b"
     misc_feature    4543..4564
                     /label=10X capture seq1
                     /note="color: #3366ff"
     polyA_signal    4565..4644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #ff0000"
     polyA_signal    4645..4686
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #ff0000"
     misc_feature    4687..4697
                     /label=PolyA
                     /note="color: #ff2549"
     misc_feature    4726..4866
                     /label=ITR
                     /note="color: #00ffff"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaca gtgtctagac tgcagagggc cctgcgtatg
      181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
      241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      601 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggatc cgccaccatg tctgcacttc
      661 tgatcctagc tcttgttgga gctgcagttg ctggcaagcc gatccctaac cctctgctgg
      721 gcctggacag cactcttgcg gccgcggcat ccagtttggg agcgcaccac atccaccatt
      781 tccatggcag cagcaagcat cattcagtgc ctattgcaat ctacaggtca ccagcatcct
      841 tgcgaggcgg acacgctggg acgacgtata tctttagcaa aggtggtgga caaattacat
      901 ataagtggcc tcctaatgac cgacccagta cacgagcaga caggctggcc atcgggttta
      961 gcactgttca gaaggaagca gtgttggtgc gtgtggacag ttcctcgggc ctgggtgact
     1021 accttgagct gcacatacac caaggaaaaa ttggagttaa atttaatgtt gggacagatg
     1081 acatcgccat tgaggagtcc aatgcaatca ttaatgatgg aaaataccat gtagtgcgtt
     1141 tcacgaggag tggtggcaat gccacgttac aggtggacag ctggccagtc atcgaacgat
     1201 accctgcatc cggaggaggt tcaggtggag gttcaggagg aggaatgggt ggaacaagca
     1261 tgagcaaagg tgaggaactg ttcaccggcg tcgtacctat tcttgtggag ctggacgggg
     1321 acgtcaatgg acacaaattc tcagttcgcg gcgagggcga gggggatgct accatcggta
     1381 agctgaccct caaattcatc tgcactacgg gcaagctgcc ggttccctgg cctactctgg
     1441 tcactacact gacttacggg gtccaatgct tctccagata tcccgaccac atgaagcggc
     1501 acgacttctt taaaagcgcc atgccggagg ggtacgtgca ggaacgaacc atttccttca
     1561 aagatgatgg taagtataag accagagctg tcgtcaagtt tgagggcgac accctcgtga
     1621 accgcatcga acttaaggga accgatttta aggaagacgg taacatcctg ggccacaagc
     1681 tggagtacaa tttcaactca cataatgtgt acattacggc agacaagcag aaaaacggga
     1741 tcaaagccaa ttttacagta cggcacaatg tcgaggacgg cagcgtccag ctggcagatc
     1801 attaccagca aaacacgcct atcggagatg ggccggtgct gcttccagat aaccattacc
     1861 tttccacaca gaccgtgctg agcaaggacc ctaacgagaa gggtggaggt tctggaggtg
     1921 gttctggagg ttccggaggg cgtcagctca caatcttcaa tagccaagca accataataa
     1981 ttggcgggaa agagcagggc cagcccttcc agggccagct ctctgggctt tactacaatg
     2041 gcttgaaagt tctgaatatg gcggcagaga acgatgccaa catcgccata gtgggaaatg
     2101 tgaggctggt cggtgaagtg ccttcctcta tgacaactga gtcgacagcc actgccatgc
     2161 agtccgagat gtccacctca atcatggaga caaccacaac cctggctacc agcacagctc
     2221 gacgaggaaa gccccccaca aaggagccta tcagccagac cacggatgat atccttgtgg
     2281 cctcggcaga gtgtcccagc gacgatgagg acattgaccc ctgtgagccg agctcaggtg
     2341 ggttagccaa ccccacccga gtgggcggac gcgaaccata cccaggctcg gcagaggtga
     2401 tccgggagtc tagcagtacc actggcatgg tggtggggat tgtcgcagca gcagctctgt
     2461 gcatcctcat cctcctctat gccatgtaca agtacaggaa ccgggatgaa ggttcaagcg
     2521 ctccaccgct cgacggagcc ggagctggcg ctggagctgg agccggagct ggcgggctag
     2581 ccaccatgga cgcacaaaca cgacgacgtg agcgtcgcgc tgagaaacaa gctcaatgga
     2641 aagctgcaaa cctcgagcca ccgctcgacg gagccggagc tggcgctgga gctggagccg
     2701 gagctggcgg tctagccacc ccaccgagcg cttatcacgt ggatgagagt cgaaactaca
     2761 tcagtaactc agcacagtcc aatggggctg tggtcaagga gaagcagccc agcagtgcga
     2821 aaagcgccaa caaaaacaag aagaacaagg ataaggagta ttatgtctga cgatatcaag
     2881 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2941 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3001 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3061 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3121 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3181 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3241 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     3301 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3361 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3421 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3481 cgataccgag cgctgctcga gctagagctc gctgatcagc ctcgactgtg ccttctagtt
     3541 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3601 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3661 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3721 ggcatgctgg ggagtgtcta gactgcagag ggccctgcgt atgagtgcaa gtgggtttta
     3781 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
     3841 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
     3901 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
     3961 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
     4021 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
     4081 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
     4141 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
     4201 tgagagcgca gcaattggag accggagaag ccctttgcca tcgccaagga gtagggcaca
     4261 gggacatctt tctttgagtg accgtctgtg caggccctgt agtccgccac agggctctga
     4321 gctgcactgg ccccggtgct ggcatctggt ggagcggacc cactcccctc acattccaca
     4381 ggcccatgga ctcactttgg tctcatgcat cctctttttc cggctggaac cttccctaag
     4441 tccaactact aaactgggct aagtccaact actaaactgg gccctgaaga agggcccata
     4501 tagggccctg aagaagggcc ctatcgagga tattatctcg aggctttaag gccggtccta
     4561 gcaaaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4621 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4681 atcttacatg ctggggaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     4741 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4801 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4861 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt cacaccgcat
     4921 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4981 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5041 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5101 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     5161 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5221 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5281 ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5341 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     5401 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     5461 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     5521 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5581 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5641 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5701 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5761 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5821 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5881 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5941 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6001 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6061 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6121 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6181 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6241 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6301 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6361 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6421 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6481 ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc
     6541 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6601 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6661 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6721 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6781 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6841 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6901 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     6961 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7021 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7081 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7141 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7201 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7261 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7321 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7381 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7441 tgctggcctt ttgctcacat gt
//
