LOCUS       sequence-318744-        6493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector containing a PIP-binding defective 
            mutant of SNX9. Also contains mCherry tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6493)
  AUTHORS   Roman-Fernandez A, Roignot J, Sandilands E, Nacke M, Mansour MA, 
            McGarry L, Shanks E, Mostov KE, Bryant DM
  TITLE     The phospholipid PI(3,4)P2 is an apical identity determinant.
  JOURNAL   Nat Commun. 2018 Nov 28;9(1):5041. doi: 10.1038/s41467-018-07464-8.
  PUBMED    30487552
REFERENCE   2  (bases 1 to 6493)
  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: "Nat 
            Commun."; date: "2018-11-28"; volume: "9(1):5041. doi"; pages: " 
            10.1038/s41467-018-07464-8"
COMMENT     Sequence Label: mCherry-C1 SNX9 RYK mutant R286A Y287A K288A
FEATURES             Location/Qualifiers
     source          1..6493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        88..391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        392..595
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     546..566
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             627..1334
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(772..790)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1040..1059
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3308..3429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3345..3364)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3399..3418
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3436..3891)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3573..3594)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3785..3804
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3918..4022
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4024..4381
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4024..4215
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4047..4067)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4232..4367
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4416..5210
                     /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             4416..5210
                     /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(4470..4489)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5080..5099
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5398..5417)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5442..5489
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5818..6406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6307..6326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
       61 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      121 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      181 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      241 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      301 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta ctttgcagta
      361 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      421 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      481 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      541 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
      601 agtgaaccgt cagatccgct agcaccatgg tgagcaaggg cgaggaggat aacatggcca
      661 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
      721 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
      781 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
      841 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
      901 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
      961 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     1021 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     1081 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     1141 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     1201 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     1261 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     1321 acgagctgta caagtccgga ctcagatcta ccatggccac caaggctcgg gtcatgtacg
     1381 actttgctgc tgagcctgga aacaatgagc tgaccgttac tgaaggagaa atcatcacag
     1441 ttacaaaccc gaatgttggt ggaggatggc tggaaggaaa gaacaacaaa ggagagcagg
     1501 ggcttgttcc tactgactac gtggaaattt taccaaacga tggaaaagat ccgttttctt
     1561 gtgggaattc agtagccgac caagctttcc tcgattccct gacagctagc acagctcaaa
     1621 ccaactcatc ttctgccaac agcaataacc aggttggtgg tggcaatgac ccctggacag
     1681 cctggaatgc tcccaaacct gggaactggg acagctccga tgcctggggc tccaggacag
     1741 acggaactag cgctcagagg aacagcagcg ccaacaactg ggacacgggc tttggccacc
     1801 cacaggccta ccaaggacca gcaactggtg atgatgacga atgggatgaa gactgggacg
     1861 accccaagtc ctcctcacct tacttcaagg attcagaacc agctgaagca gggggcatcc
     1921 agcggggaaa cagtcgtgcg ggtgcctcct ccatgaagct gccccttaac aaatttcctg
     1981 gatttgcaaa acctggcatg gagcagtact tgttggccaa gcagctagca aaacccaaag
     2041 agaaaatcgc catcatcgtt ggagattatg gccctatgtg ggtttatcct acctctacgt
     2101 ttgactgtgt ggtagctgat ccccggaaag gttccaaaat gtacggtctg aagagctaca
     2161 ttgagtatca gctaacaccc actaacacta accgatctgt aaaccacgca gctgcgcact
     2221 ttgattggtt atatgagcgt ctcctggtta agtttgggtc tgccattccg atcccttctc
     2281 ttccagacaa gcaggtcaca ggtcgctttg aagaggaatt tatcaaaatg cggatggagc
     2341 ggctccaggc ctggatgacc aggatgtgcc ggcacccggt ggtgtcagaa agtgaagttt
     2401 tccagcagtt cctgaatttc cgagatgaga aggaatggaa gactggaaag aggaaggctg
     2461 agaaagacga actggtggga gtgatgatat tttccaccat ggaaccagag gctcctgact
     2521 tggatttgat agaaatagaa cagaagtgtg acgctgtggg gaagttcacc aaggccatgg
     2581 atgacggcgt gaaggagctg ctgacggtgg ggcaggagca ctggaagcgc tgcacagggc
     2641 ccttacccaa ggagtatcag aagataggga aggctctgca gagcttagcg gcagtgttca
     2701 gctccagcgg ttatcaagga gagaccgacc ttaacgatgc aatcacagaa gctgggaaga
     2761 cgtatgagga gatcgccagc cttgtggcag agcagccaaa gaaggacctc cacttcctga
     2821 tggagtgcaa ccatgagtac aaaggctttc tcggctgctt cccggacatc attggtgccc
     2881 acaagggagc aatagaaaaa gtgaaagaaa gtgataagct ggttgcaact agtaaaatta
     2941 ctccccaaga caagcagacc atggtgaaga gagttggcac catgtcttac gctctgcaag
     3001 ctgagatgaa tcactttcac agtaaccgga tctatgatta caacagtgtc atccgcctct
     3061 acctggagca gcaagtgcag ttctatgaaa caatcgcaga aaagctgagg caggccctca
     3121 gccgcttccc ggttatgggt tcttcaggat catccgggtc gacggtaccg cgggcccggg
     3181 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     3241 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3301 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3361 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3421 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3481 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3541 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3601 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3661 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3721 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3781 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3841 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3901 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3961 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4021 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4081 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4141 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4201 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4261 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4321 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4381 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4441 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4501 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4561 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4621 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4681 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4741 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4801 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4861 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4921 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4981 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5041 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5101 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5161 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5221 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5281 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5341 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5401 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     5461 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5521 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5581 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5641 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5701 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5761 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5821 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5881 ggtggtttgt ttgtcggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5941 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6001 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6061 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6121 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6181 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6241 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6301 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6361 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6421 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6481 atcccctgat tct
//
