LOCUS       sequence-458082-        6333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing HRP on the cell surface of astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6333)
  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 6333)
  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: GfaABC1D HRP-TM
FEATURES             Location/Qualifiers
     source          1..6333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     intron          1239..1371
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1354..1373
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1416..1434
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     1461..1520
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1524..1550
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2502..2531
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2535..2681
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    2706..3294
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2759..2779)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3177..3188)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3358..3479
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3369..3388)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3423..3442
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3641..3770
                     /label=AAV2 ITR
     primer_bind     complement(3789..3806)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3789..3805)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3798..3820)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3947..4402
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4034..4053)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4244..4265
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4428..4532
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4533..4601,4602..5393)
                     /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"
     rep_origin      5564..6152
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6053..6072
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatctaa catatcctgg tgtggagtag gggacgctgc
      481 tctgacagag gctcgggggc ctgagctggc tctgtgagct ggggaggagg cagacagcca
      541 ggccttgtct gcaagcagac ctggcagcat tgggctggcc gccccccagg gcctcctctt
      601 catgcccagt gaatgactca ccttggcaca gacacaatgt tcggggtggg cacagtgcct
      661 gcttcccgcc gcaccccagc ccccctcaaa tgccttccga gaagcccatt gagcaggggg
      721 cttgcattgc accccagcct gacagcctgg catcttggga taaaagcagc acagccccct
      781 aggggctgcc cttgctgtgt ggcgccaccg gcggtggaga acaaggctct attcagcctg
      841 tgcccaggaa aggggatcag gggatgccca ggcatggaca gtgggtggca gggggggaga
      901 ggagggctgt ctgcttccca gaagtccaag gacacaaatg ggtgagggga gagctctccc
      961 catagctggg ctgcggccca accccacccc ctcaggctat gccagggggt gttgccaggg
     1021 gcacccgggc atcgccagtc tagcccactc cttcataaag ccctcgcatc ccaggagcga
     1081 gcagagccag agcaggttgg agaggagacg catcacctcc gctgctcgca agctttattg
     1141 cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca acagtctcga
     1201 acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
     1261 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     1321 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
     1381 cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta taggctagcc
     1441 tcgagctcgg atccaccacc atggagacag acacactcct gctatgggta ctgctgctct
     1501 gggttccagg ttccactggt gactatccat atgatgttcc agattatgct ggggcccagc
     1561 agttaacccc tacattctac gacaatagct gtcccaacgt gtccaacatc gttcgcgaca
     1621 caatcgtcaa cgagctcaga tccgatccca ggatcgctgc ttcaatatta cgtctgcact
     1681 tccatgactg cttcgtgaat ggttgcgacg ctagcatatt actggacaac accaccagtt
     1741 tccgcactga aaaggatgca ttcgggaacg ctaacagcgc caggggcttt ccagtgatcg
     1801 atcgcatgaa ggctgccgtt gagtcagcat gcccacgaac agtcagttgt gcagacctgc
     1861 tgactatagc tgcgcaacag agcgtgactc ttgcaggcgg accgtcctgg agagtgccgc
     1921 tcggtcgacg tgactcccta caggcattcc tagatctggc caacgccaac ttgcctgctc
     1981 cattcttcac cctgccccag ctgaaggata gctttagaaa cgtgggtctg aatcgctcga
     2041 gtgaccttgt ggctctgtcc ggaggacaca catttggaaa gaaccagtgt aggttcatca
     2101 tggataggct ctacaatttc agcaacactg ggttacctga ccccacgctg aacactacgt
     2161 atctccagac actgagaggc ttgtgcccac tgaatggcaa cctcagtgca ctagtggact
     2221 ttgatctgcg gaccccaacc atcttcgata acaagtacta tgtgaatcta gaggagcaga
     2281 aaggcctgat acagagtgat caagaactgt ttagcagtcc aaacgccact gacaccatcc
     2341 cactggtgag aagttttgct aactctactc aaaccttctt taacgccttc gtggaagcca
     2401 tggaccgtat gggtaacatt acccctctga cgggtaccca aggccagatt cgtctgaact
     2461 gcagagtggt caacagcaac tctccgcggc tgcaggtcga cgaacaaaaa ctcatctcag
     2521 aagaggatct gaatgctgtg ggccaggaca cgcaggaggt catcgtggtg ccacactcct
     2581 tgccctttaa ggtggtggtg atctcagcca tcctggccct ggtggtgctc accatcatct
     2641 cccttatcat cctcatcatg ctttggcaga agaagccacg ttagggatcc aattcgctta
     2701 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2761 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2821 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2881 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2941 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3001 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3061 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3121 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3181 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3241 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3301 accggaattc tgcagatatc tagagtcgac ccgggcggcc gcttcgagca gacatgataa
     3361 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     3421 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     3481 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     3541 aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc
     3601 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     3661 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3721 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ccttaattaa
     3781 cctaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     3841 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     3901 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     3961 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4021 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4081 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4141 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4201 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4261 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4321 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4381 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4441 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4501 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4561 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4621 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4681 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4741 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4801 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4861 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4921 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4981 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5041 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5101 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5161 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5221 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5281 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5341 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5401 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5461 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5521 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5581 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5641 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5701 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5761 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5821 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5881 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5941 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6001 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6061 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6121 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6181 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6241 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     6301 agcgcagcga gtcagtgagc gaggaagcgg aag
//
