LOCUS       sequence-321957-        6314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mouse SNX27 cDNA with point mutation to resist siRNA against SNX27 
            and with a point mutation H112A C-terminally tagged with GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6314)
  AUTHORS   Halff EF, Szulc BR, Lesept F, Kittler JT
  TITLE     SNX27-Mediated Recycling of Neuroligin-2 Regulates Inhibitory 
            Signaling.
  JOURNAL   Cell Rep. 2019 Nov 26;29(9):2599-2607.e6. doi: 
            10.1016/j.celrep.2019.10.096.
  PUBMED    31775031
REFERENCE   2  (bases 1 to 6314)
  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"; doi: 
            "10.1016/j.celrep.2019.10"; journalName: "Cell Rep"; date: 
            "2019-11-26- 26"; volume: "29"; issue: "9"; pages: "2599-2607.e6"
COMMENT     Sequence Label: pEGFP-N1-mSNX27-H112A RNAi-resistant
FEATURES             Location/Qualifiers
     source          1..6314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        37..340
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        341..544
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     495..515
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2230..2239
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2236..2238,2239..2241,2242..2955)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2236..2238,2239..2241,2242..2952)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2281..2302)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2542..2561)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2889..2910
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3078..3199
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3115..3134)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3169..3188
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3206..3661)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3343..3364)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3555..3574
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3688..3792
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3794..4151
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3794..3985
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3817..3837)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4002..4137
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4186..4980
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4186..4980
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4240..4259)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4850..4869
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5168..5187)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5212..5259
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5588..6176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6077..6096
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
       61 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      121 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      181 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      241 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      301 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      361 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      421 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      481 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      541 agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct cgagctcaag
      601 cttatggcgg acgaggacgg ggaaggtatt cacccctcag ccccgcaccg gaatggaggc
      661 ggcggcgggg ggtctgggct ccactgcgcc gggaacggtg gcgggggagg cggcggcccg
      721 cgggtcgtgc gcattgtcaa gtccgaatcc ggctacggct tcaacgtgcg gggccaagtg
      781 agcgagggcg ggcaactgcg gagcatcaac ggggagctgt acgcgccgct gcagcatgtg
      841 agcgccgtgc tgcccggggg ggcggccgac cgggccgggg tgcgcaaagg ggaccgcatc
      901 ctggaggtga acggcgtgaa tgttgagggg gcgacagcca agcaggtggt ggacctgatc
      961 cgagcaggcg agaaggaatt gatcttgaca gtgttgtctg tacctcctca tgaggctgat
     1021 aacctagatc ccagtgacga ctccttggga caatcatttt atgattacac agaaaagcaa
     1081 gcagtgccca tatcggtccc cacatacaaa catgtggagc agaatggtga gaagtttgtg
     1141 gtatacaatg tttacatggc tgggaggcag ctgtgttcta agcggtaccg ggagtttgcc
     1201 atactgcacc aaaacctgaa gagagaattt gccaacttta catttcctcg acttccagga
     1261 aagtggccat tctcattatc agaacaacaa ttggatgccc ggcgtcgtgg gttggaagaa
     1321 tatctagaaa aggtgtgttc aatccgagtc attggtgaga gtgacatcat gcaggaattc
     1381 ctgtctgagt ctgatgagaa ttacaatggt gtgtctgatg tagaactgag agtagcacta
     1441 ccggatggaa caacggttac tgtcagagtt aagaagaaca gtactacaga ccaagtgtac
     1501 caggctattg cagcaaaggt tggcatggac agtacaacag tgaattactt tgccttattt
     1561 gaagtgatca atcattcctt tgtacgtaag ctggcaccta atgaatttcc tcataaactc
     1621 tacgtccaga actatacatc agctgtgcca ggcacttgcc tgaccattcg caagtggctc
     1681 tttacaacag aagaagaagt cctcttaaat gacaatgacc ttgctgttac gtacttcttt
     1741 catcaggctg ttgatgatgt aaagaaaggt tacattaaag cagaagagaa atcttaccag
     1801 ttacagaagc tccatgaaca aaggaagatg gtcatgtacc tcaacatgct aaggacctgc
     1861 gagggctaca atgaaatcat cttccctcac tgtgcctgtg actcgaggag gaagggacat
     1921 gttatcacag ccatcagcat cacgcacttt aaacttcatg catgcactga agaagggcag
     1981 ctggagaacc aagtgatcgc attcgaatgg gatgagatgc agcgatggga cacagatgaa
     2041 gaaggaatgg ccttctgttt tgaatatgcg cgaggagaga agaagcctcg atgggttaaa
     2101 atcttcacac catatttcaa ttacatgcat gaatgctttg agagggtgtt ctgcgagctc
     2161 aagtggagaa aagagaatat tttccagatg gcgaggtcac agcagaggga tgtggccacc
     2221 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2281 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     2341 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     2401 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     2461 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     2521 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2581 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2641 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     2701 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     2761 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2821 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     2881 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2941 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     3001 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3061 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3121 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3181 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3241 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3301 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3361 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3421 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3481 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3541 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3601 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3661 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3721 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3781 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3841 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3901 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3961 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4021 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4081 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4141 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4201 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4261 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4321 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4381 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4441 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4501 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4561 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4621 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4681 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4741 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4801 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4861 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4921 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4981 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5041 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5101 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     5161 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5221 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5281 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5341 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     5401 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     5461 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5521 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5581 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5641 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5701 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5761 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5821 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5881 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5941 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6001 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6061 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6121 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6181 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6241 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     6301 tagtaatcaa ttac
//
