LOCUS       sequence-321951-        6314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mouse SNX27 cDNA with point mutation to resist siRNA against SNX27 
            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 RNAi-resistant
FEATURES             Location/Qualifiers
     source          1..6314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        366..569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     520..540
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2255..2264
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2261..2263,2264..2266,2267..2980)
                     /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(2261..2263,2264..2266,2267..2977)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2306..2327)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2567..2586)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2914..2935
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3103..3224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3140..3159)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3194..3213
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3231..3686)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3368..3389)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3580..3599
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3713..3817
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3819..4176
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3819..4010
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3842..3862)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4027..4162
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4211..5005
                     /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             4211..5005
                     /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(4265..4284)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4875..4894
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5193..5212)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5237..5284
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5613..6201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6102..6121
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
       61 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      241 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      301 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      361 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      421 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      481 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      541 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      601 accggactca gatctcgagc tcaagcttat ggcggacgag gacggggaag gtattcaccc
      661 ctcagccccg caccggaatg gaggcggcgg cggggggtct gggctccact gcgccgggaa
      721 cggtggcggg ggaggcggcg gcccgcgggt cgtgcgcatt gtcaagtccg aatccggcta
      781 cggcttcaac gtgcggggcc aagtgagcga gggcgggcaa ctgcggagca tcaacgggga
      841 gctgtacgcg ccgctgcagc atgtgagcgc cgtgctgccc gggggggcgg ccgaccgggc
      901 cggggtgcgc aaaggggacc gcatcctgga ggtgaacggc gtgaatgttg agggggcgac
      961 acacaagcag gtggtggacc tgatccgagc aggcgagaag gaattgatct tgacagtgtt
     1021 gtctgtacct cctcatgagg ctgataacct agatcccagt gacgactcct tgggacaatc
     1081 attttatgat tacacagaaa agcaagcagt gcccatatcg gtccccacat acaaacatgt
     1141 ggagcagaat ggtgagaagt ttgtggtata caatgtttac atggctggga ggcagctgtg
     1201 ttctaagcgg taccgggagt ttgccatact gcaccaaaac ctgaagagag aatttgccaa
     1261 ctttacattt cctcgacttc caggaaagtg gccattctca ttatcagaac aacaattgga
     1321 tgcccggcgt cgtgggttgg aagaatatct agaaaaggtg tgttcaatcc gagtcattgg
     1381 tgagagtgac atcatgcagg aattcctgtc tgagtctgat gagaattaca atggtgtgtc
     1441 tgatgtagaa ctgagagtag cactaccgga tggaacaacg gttactgtca gagttaagaa
     1501 gaacagtact acagaccaag tgtaccaggc tattgcagca aaggttggca tggacagtac
     1561 aacagtgaat tactttgcct tatttgaagt gatcaatcat tcctttgtac gtaagctggc
     1621 acctaatgaa tttcctcata aactctacgt ccagaactat acatcagctg tgccaggcac
     1681 ttgcctgacc attcgcaagt ggctctttac aacagaagaa gaagtcctct taaatgacaa
     1741 tgaccttgct gttacgtact tctttcatca ggctgttgat gatgtaaaga aaggttacat
     1801 taaagcagaa gagaaatctt accagttaca gaagctccat gaacaaagga agatggtcat
     1861 gtacctcaac atgctaagga cctgcgaggg ctacaatgaa atcatcttcc ctcactgtgc
     1921 ctgtgactcg aggaggaagg gacatgttat cacagccatc agcatcacgc actttaaact
     1981 tcatgcatgc actgaagaag ggcagctgga gaaccaagtg atcgcattcg aatgggatga
     2041 gatgcagcga tgggacacag atgaagaagg aatggccttc tgttttgaat atgcgcgagg
     2101 agagaagaag cctcgatggg ttaaaatctt cacaccatat ttcaattaca tgcatgaatg
     2161 ctttgagagg gtgttctgcg agctcaagtg gagaaaagag aatattttcc agatggcgag
     2221 gtcacagcag agggatgtgg ccaccccacc ggtcgccacc atggtgagca agggcgagga
     2281 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2341 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     2401 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     2461 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2521 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2581 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2641 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2701 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2761 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2821 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     2881 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2941 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg actctagatc
     3001 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3061 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3121 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3181 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     3241 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3301 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     3361 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     3421 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     3481 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     3541 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     3601 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3661 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     3721 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3781 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     3841 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3901 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3961 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4021 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4081 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4141 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     4201 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4261 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4321 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4381 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4441 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4501 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4561 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4621 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4681 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     4741 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4801 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4861 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4921 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4981 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5041 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5101 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5161 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     5221 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     5281 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     5341 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     5401 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     5461 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5521 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5581 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5641 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5701 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5761 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5821 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5881 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5941 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6001 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6061 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6121 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6181 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6241 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6301 attaccgcca tgca
//
