LOCUS       sequence-458080-        5953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing HRP on the cell surface under CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5953)
  AUTHORS   Wu L, Pandey V, Casha VH, Qu Z, Jami-Alahmadi Y, Gradinaru V, 
            Wohlschlegel JA, Khakh BS
  TITLE     The cell-surface shared proteome of astrocytes and neurons and the 
            molecular foundations of their multicellular interactions.
  JOURNAL   Neuron. 2025 Jun 5:S0896-6273(25)00366-6. doi: 
            10.1016/j.neuron.2025.05.019.
  PUBMED    40499536
REFERENCE   2  (bases 1 to 5953)
  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: "Neuron. 
            2025 Jun 5:S0896-6273(25)00366-6. doi: 
            10.1016/j.neuron.2025.05.019."
COMMENT     Sequence Label: CAG HRP-TM
FEATURES             Location/Qualifiers
     source          1..5953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        441..722
                     /label=chicken beta-actin promoter
     primer_bind     1001..1023
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1085..1104
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1143..1152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1149..1208
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1212..1238
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2190..2219
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2223..2369
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    2397..2985
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2450..2470)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2868..2879)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3028..3149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3039..3058)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3093..3112
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3311..3440
                     /label=AAV2 ITR
     rep_origin      complement(3677..4265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3757..3776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4436..5227,5228..5296))
                     /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     5059..5078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5297..5401)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5427..5882)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5564..5585)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5776..5795
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg ggggggcgcg cgccrggsgg ggcggggcgg ggcgaggggc
      601 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      661 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      721 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      781 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      841 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      901 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggctgtccg
      961 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1021 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1081 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt ggatccggta
     1141 ccgccaccat ggagacagac acactcctgc tatgggtact gctgctctgg gttccaggtt
     1201 ccactggtga ctatccatat gatgttccag attatgctgg ggcccagcag ttaaccccta
     1261 cattctacga caatagctgt cccaacgtgt ccaacatcgt tcgcgacaca atcgtcaacg
     1321 agctcagatc cgatcccagg atcgctgctt caatattacg tctgcacttc catgactgct
     1381 tcgtgaatgg ttgcgacgct agcatattac tggacaacac caccagtttc cgcactgaaa
     1441 aggatgcatt cgggaacgct aacagcgcca ggggctttcc agtgatcgat cgcatgaagg
     1501 ctgccgttga gtcagcatgc ccacgaacag tcagttgtgc agacctgctg actatagctg
     1561 cgcaacagag cgtgactctt gcaggcggac cgtcctggag agtgccgctc ggtcgacgtg
     1621 actccctaca ggcattccta gatctggcca acgccaactt gcctgctcca ttcttcaccc
     1681 tgccccagct gaaggatagc tttagaaacg tgggtctgaa tcgctcgagt gaccttgtgg
     1741 ctctgtccgg aggacacaca tttggaaaga accagtgtag gttcatcatg gataggctct
     1801 acaatttcag caacactggg ttacctgacc ccacgctgaa cactacgtat ctccagacac
     1861 tgagaggctt gtgcccactg aatggcaacc tcagtgcact agtggacttt gatctgcgga
     1921 ccccaaccat cttcgataac aagtactatg tgaatctaga ggagcagaaa ggcctgatac
     1981 agagtgatca agaactgttt agcagtccaa acgccactga caccatccca ctggtgagaa
     2041 gttttgctaa ctctactcaa accttcttta acgccttcgt ggaagccatg gaccgtatgg
     2101 gtaacattac ccctctgacg ggtacccaag gccagattcg tctgaactgc agagtggtca
     2161 acagcaactc tccgcggctg caggtcgacg aacaaaaact catctcagaa gaggatctga
     2221 atgctgtggg ccaggacacg caggaggtca tcgtggtgcc acactccttg ccctttaagg
     2281 tggtggtgat ctcagccatc ctggccctgg tggtgctcac catcatctcc cttatcatcc
     2341 tcatcatgct ttggcagaag aagccacgtt aggaattcga tatcaagctt atcgataatc
     2401 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2461 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2521 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2581 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2641 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2701 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2761 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     2821 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     2881 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     2941 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac
     3001 ccgggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3061 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3121 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3181 aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3241 aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc
     3301 attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3361 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3421 gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3481 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3541 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3601 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3661 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3721 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3781 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3841 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3901 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3961 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4021 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4081 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4141 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4201 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4261 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4321 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4381 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4441 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4501 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4561 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4621 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4681 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4741 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4801 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4861 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4921 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4981 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5041 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5101 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5161 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5221 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5281 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5341 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5401 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     5461 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5521 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5581 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5641 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5701 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5761 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5821 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5881 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5941 gctattacgc cag
//
