LOCUS       sequence-318748-        7479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector containing INPP4B. Also contains mCherry
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7479)
  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 7479)
  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: pmCherry-C1-INPP4B WT
FEATURES             Location/Qualifiers
     source          1..7479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        302..505
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     456..476
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             534..1241
                     /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(679..697)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     947..966
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4204..4325
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4241..4260)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4295..4314
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4332..4787)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4469..4490)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4681..4700
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4814..4918
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4920..5277
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4920..5111
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4943..4963)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5128..5263
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5312..6106
                     /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             5312..6106
                     /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(5366..5385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5976..5995
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6294..6313)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6338..6385
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6714..7302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7203..7222
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(7477..7479,1..301)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
       61 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      121 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      181 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      241 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      301 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      361 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      421 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      481 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcatggtga
      541 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
      601 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
      661 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
      721 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
      781 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
      841 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
      901 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
      961 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     1021 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     1081 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     1141 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     1201 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtccggactc agatctcgag
     1261 ctcaagcttc gaattctgca gtcgacggta ccatggaaat taaagaggaa ggggcatcag
     1321 aagaagggca gcactttctt cctacagccc aggccaatga tcccggggac tgtcagttca
     1381 caagtatcca gaagactcca aatgaaccgc agttggaatt catccttgca tgcaaggatc
     1441 tcgtggctcc tgtccgtgat cgtaaactga atacactggt gcagatctcc gtaatccacc
     1501 ccgtggagca gagtctgaca agatactcca gcaccgaaat tgtggaggga acaagggacc
     1561 cactgttttt gactggtgtc acattcccat ctgagtatcc catctatgag gagaccaaaa
     1621 taaaactaac agtctatgat gtcaaggata agtctcatga caccgttcga accagtgtcc
     1681 taccagaaca taaggatccc ccgccagaag ttgggcgaag tttcttgggc tatgccagtt
     1741 ttaaagtggg agagctgctg aagtcaaagg agcaattgct ggtcctgagc ctgagaactt
     1801 cagatggtgg caaagtggtt ggcaccatag aagtcagtgt cgtgaagatg ggggagattg
     1861 aggatgggga agccgaccac atcaccacag atgtacaggg acaaaagtgt gccctggtat
     1921 gtgaatgtac agccccggaa agtgtgagcg gaaaagataa cttacctttt ttgaattcag
     1981 tgttaaagaa cccagtatgt aaattatata gatttcccac atctgacaat aagtggatgc
     2041 gaattcgaga gcagatgtca gagagcattc tttcctttca tattcctaag gaattgattt
     2101 cccttcacat taaagaagat ttgtgcagaa accaggagat aaaagaactt ggtgagcttt
     2161 ctccacattg ggacaatctg cgaaaaaatg tccttactca ctgtgatcaa atggtgaata
     2221 tgtaccaaga cattctgaca gaacttagca aggaaacagg gtcctctttc aaatcaagca
     2281 gcagcaaagg agagaaaaca ttagaatttg ttccaataaa tctacatctg caaagaatgc
     2341 aggtacacag ccctcacttg aaagatgctc tctacgatgt catcactgtg ggagccccag
     2401 ctgcccattt tcagggattt aagaatggtg gtcttcggaa gctactccat agatttgaaa
     2461 cagaaagaag aaataccgga taccagttta tttactattc acctgaaaac acagccaaag
     2521 caaaggaagt tctcagcaac atcaatcaac tacaacctct tatagcaacc catgcagacc
     2581 tactgcttaa ttctgcaagc cagcattctc cagacagctt gaagaattct ttaaagatgc
     2641 tttcagaaaa aacagagctt tttgtacatg ccttcaagga tcaacttgtc aggagtgctc
     2701 ttttagcact ctacactgca aggccaggag gcattcttaa gaagccaccc tctcctaaga
     2761 gcagcacaga ggagagcagt ccccaagacc aacccccagt gatgagaggg caggactcca
     2821 taccacatca ttcagactat gatgaggaag agtgggacag ggtgtgggcc aatgtgggga
     2881 agagcctgaa ctgcattatt gctatggtgg acaaactgat tgaaagagat ggtggcagtg
     2941 aaggcagtgg cggcaacaat gatggagaaa aggaaccttc attaacagat gccattccct
     3001 ctcacccaag agaggactgg tatgaacagt tgtatcccct catccttacc ctgaaggact
     3061 gcatgggaga agtggtgaac cgagccaagc agtccctgac atttgtgctc cttcaggaac
     3121 ttgcgtacag cttgccccag tgtctgatgc tgacgctaag aagagacatc gtcttcagcc
     3181 aagcacttgc tggattggtt tgtggtttta tcatcaaatt acagacaagt ctgtatgacc
     3241 caggcttcct acagcagctt cacacagtgg ggttgatagt acaatatgaa ggactgctaa
     3301 gtacatacag cgatgaaatt ggaatgctag aggacatggc cgttggcatt tccgatttaa
     3361 agaaagttgc atttaaaata attgaagcca aatccaatga tgtgttgcca gttataacag
     3421 gaagacgaga acattacgtg gtagaggtca agcttccagc cagaatgttt gagtcactac
     3481 ctctacagat taaagaagga cagttgcttc atgtgtatcc agtacttttt aatgttggaa
     3541 tcaatgaaca gcaaactctg gctgaaaggt ttggagatgt ctctttgcaa gaaagtatta
     3601 atcaggaaaa cttcgaactt ctacaagaat attacaagat atttatggaa aagatgcctc
     3661 ctgattatat ttcacatttt caggaacaaa atgatttaaa agcattgcta gaaaatctcc
     3721 ttcaaaatat ccaatccaaa aaaagaaaga atgtagaaat tatgtggctg gctgcaacga
     3781 tttgccgcaa actgaatggt attcgtttca cctgttgtaa aagtgccaaa gacaggacat
     3841 cgatgtcagt gacacttgaa caatgctcaa tcttgagaga tgagcaccag ttacacaagg
     3901 acttctttat ccgagcgctg gattgcatga gaagagaagg atgccgcata gagaatgtac
     3961 tgaagaatat caaatgcaga aagtatgctt tcaacatgct acagctgatg gctttcccca
     4021 agtactacag acctccagag gggacttatg gaaaagctga cacctaaggg cccgggatcc
     4081 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     4141 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     4201 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4261 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4321 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     4381 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     4441 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     4501 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     4561 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     4621 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     4681 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4741 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     4801 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4861 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     4921 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4981 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5041 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5101 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5161 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     5221 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     5281 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     5341 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5401 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5461 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     5521 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5581 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5641 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5701 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5761 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5821 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5881 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5941 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     6001 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6061 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     6121 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     6181 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     6241 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     6301 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     6361 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     6421 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     6481 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     6541 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     6601 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6661 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6721 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6781 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6841 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6901 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6961 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7021 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7081 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7141 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7201 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7261 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7321 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7381 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     7441 ggggtcatta gttcatagcc catatatgga gttccgcgt
//
