LOCUS       sequence-318809-        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 KR mutant K267N R327N
FEATURES             Location/Qualifiers
     source          1..6493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        89..392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        393..596
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     547..567
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             628..1335
                     /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(773..791)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1041..1060
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3309..3430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3346..3365)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3400..3419
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3437..3892)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3574..3595)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3786..3805
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3919..4023
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4025..4382
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4025..4216
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4048..4068)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4233..4368
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4417..5211
                     /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             4417..5211
                     /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(4471..4490)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5081..5100
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5399..5418)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5443..5490
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5819..6407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6308..6327
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
       61 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      121 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      181 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      241 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      301 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      361 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      421 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      481 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      541 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
      601 tagtgaaccg tcagatccgc tagcaccatg gtgagcaagg gcgaggagga taacatggcc
      661 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
      721 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      781 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
      841 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
      901 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
      961 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     1021 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     1081 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     1141 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     1201 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     1261 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     1321 gacgagctgt acaagtccgg actcagatct accatggcca ccaaggctcg ggtcatgtac
     1381 gactttgctg ctgagcctgg aaacaatgag ctgaccgtta ctgaaggaga aatcatcaca
     1441 gttacaaacc cgaatgttgg tggaggatgg ctggaaggaa agaacaacaa aggagagcag
     1501 gggcttgttc ctactgacta cgtggaaatt ttaccaaacg atggaaaaga tccgttttct
     1561 tgtgggaatt cagtagccga ccaagctttc ctcgattccc tgacagctag cacagctcaa
     1621 accaactcat cttctgccaa cagcaataac caggttggtg gtggcaatga cccctggaca
     1681 gcctggaatg ctcccaaacc tgggaactgg gacagctccg atgcctgggg ctccaggaca
     1741 gacggaacta gcgctcagag gaacagcagc gccaacaact gggacacggg ctttggccac
     1801 ccacaggcct accaaggacc agcaactggt gatgatgacg aatgggatga agactgggac
     1861 gaccccaagt cctcctcacc ttacttcaag gattcagaac cagctgaagc agggggcatc
     1921 cagcggggaa acagtcgtgc gggtgcctcc tccatgaagc tgccccttaa caaatttcct
     1981 ggatttgcaa aacctggcat ggagcagtac ttgttggcca agcagctagc aaaacccaaa
     2041 gagaaaatcg ccatcatcgt tggagattat ggccctatgt gggtttatcc tacctctacg
     2101 tttgactgtg tggtagctga tccccggaaa ggttccaaaa tgtacggtct gaatagctac
     2161 attgagtatc agctaacacc cactaacact aaccgatctg taaaccacag atataagcac
     2221 tttgattggt tatatgagcg tctcctggtt aagtttgggt ctgccattcc gatcccttct
     2281 cttccagaca agcaggtcac aggtcgcttt gaagaggaat ttatcaaaat gaatatggag
     2341 cggctccagg cctggatgac caggatgtgc cggcacccgg tggtgtcaga aagtgaagtt
     2401 ttccagcagt tcctgaattt ccgagatgag aaggaatgga agactggaaa gaggaaggct
     2461 gagaaagacg aactggtggg agtgatgata ttttccacca tggaaccaga ggctcctgac
     2521 ttggatttga tagaaataga acagaagtgt gacgctgtgg ggaagttcac caaggccatg
     2581 gatgacggcg tgaaggagct gctgacggtg gggcaggagc actggaagcg ctgcacaggg
     2641 cccttaccca aggagtatca gaagataggg aaggctctgc agagcttagc ggcagtgttc
     2701 agctccagcg gttatcaagg agagaccgac cttaacgatg caatcacaga agctgggaag
     2761 acgtatgagg agatcgccag ccttgtggca gagcagccaa agaaggacct ccacttcctg
     2821 atggagtgca accatgagta caaaggcttt ctcggctgct tcccggacat cattggtgcc
     2881 cacaagggag caatagaaaa agtgaaagaa agtgataagc tggttgcaac tagtaaaatt
     2941 actccccaag acaagcagac catggtgaag agagttggca ccatgtctta cgctctgcaa
     3001 gctgagatga atcactttca cagtaaccgg atctatgatt acaacagtgt catccgcctc
     3061 tacctggagc agcaagtgca gttctatgaa acaatcgcag aaaagctgag gcaggccctc
     3121 agccgcttcc cggttatggg ttcttcagga tcatccgggt cgacggtacc gcgggcccgg
     3181 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     3241 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3301 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3361 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3421 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3481 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3541 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3601 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3661 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3721 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3781 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3841 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3901 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3961 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4021 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4081 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4141 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4201 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4261 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4321 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4381 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4441 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4501 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4561 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4621 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4681 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4741 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4801 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4861 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4921 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4981 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5041 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5101 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5161 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5221 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5281 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5341 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     5401 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5461 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     5521 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5581 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     5641 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5701 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5761 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5821 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5881 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5941 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6001 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6061 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6121 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6181 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6241 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6301 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6361 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6421 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6481 tatcccctga ttc
//
