LOCUS       sequence-458078-        6583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing HRP on the cell surface of mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6583)
  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 6583)
  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: pcDNA3.1-HRP-TM
FEATURES             Location/Qualifiers
     source          1..6583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     895..954
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             958..984
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1936..1965
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1969..2115
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     promoter        complement(2137..2155)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2138..2155)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2175..2192)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2181..2405
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2451..2879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2538..2557)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2748..2769
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2888..2908)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2893..3222
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3073..3208
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3135..3154
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3288..4082
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3342..3361)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3952..3971
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4256..4377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4293..4312)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4347..4366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4426..4442)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4426..4442)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4439..4461)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4450..4466
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4474..4480,4481..4498,4499..4504))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4519..4540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4657..4674)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4828..5416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4908..4927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5587..6378,6379..6447))
                     /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     6210..6229
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6448..6552)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttatggag
      901 acagacacac tcctgctatg ggtactgctg ctctgggttc caggttccac tggtgactat
      961 ccatatgatg ttccagatta tgctggggcc cagcagttaa cccctacatt ctacgacaat
     1021 agctgtccca acgtgtccaa catcgttcgc gacacaatcg tcaacgagct cagatccgat
     1081 cccaggatcg ctgcttcaat attacgtctg cacttccatg actgcttcgt gaatggttgc
     1141 gacgctagca tattactgga caacaccacc agtttccgca ctgaaaagga tgcattcggg
     1201 aacgctaaca gcgccagggg ctttccagtg atcgatcgca tgaaggctgc cgttgagtca
     1261 gcatgcccac gaacagtcag ttgtgcagac ctgctgacta tagctgcgca acagagcgtg
     1321 actcttgcag gcggaccgtc ctggagagtg ccgctcggtc gacgtgactc cctacaggca
     1381 ttcctagatc tggccaacgc caacttgcct gctccattct tcaccctgcc ccagctgaag
     1441 gatagcttta gaaacgtggg tctgaatcgc tcgagtgacc ttgtggctct gtccggagga
     1501 cacacatttg gaaagaacca gtgtaggttc atcatggata ggctctacaa tttcagcaac
     1561 actgggttac ctgaccccac gctgaacact acgtatctcc agacactgag aggcttgtgc
     1621 ccactgaatg gcaacctcag tgcactagtg gactttgatc tgcggacccc aaccatcttc
     1681 gataacaagt actatgtgaa tctagaggag cagaaaggcc tgatacagag tgatcaagaa
     1741 ctgtttagca gtccaaacgc cactgacacc atcccactgg tgagaagttt tgctaactct
     1801 actcaaacct tctttaacgc cttcgtggaa gccatggacc gtatgggtaa cattacccct
     1861 ctgacgggta cccaaggcca gattcgtctg aactgcagag tggtcaacag caactctccg
     1921 cggctgcagg tcgacgaaca aaaactcatc tcagaagagg atctgaatgc tgtgggccag
     1981 gacacgcagg aggtcatcgt ggtgccacac tccttgccct ttaaggtggt ggtgatctca
     2041 gccatcctgg ccctggtggt gctcaccatc atctccctta tcatcctcat catgctttgg
     2101 cagaagaagc cacgttagta gtctagaggg ccctattcta tagtgtcacc taaatgctag
     2161 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2221 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2281 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2341 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2401 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2461 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2521 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2581 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2641 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2701 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2761 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2821 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     2881 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2941 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3001 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3061 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3121 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3181 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc ggagcttgta
     3241 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3301 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3361 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3421 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     3481 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3541 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3601 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3661 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3721 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3781 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     3841 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     3901 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     3961 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4021 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4081 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4141 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4201 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4261 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4321 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4381 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     4441 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4501 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4561 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4621 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4681 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4741 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4801 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4861 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4921 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4981 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5041 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5101 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5161 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5221 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5281 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5341 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5401 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5461 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5521 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5581 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5641 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5701 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5761 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5821 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5881 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5941 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6001 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6061 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6121 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6181 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6241 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6301 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6361 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6421 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6481 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6541 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
