LOCUS       sequence-318752-        7479 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression vector containing a phosphatase dead mutant of 
            INPP4B (C842A). 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 Apr 14, 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 C842A mutant
FEATURES             Location/Qualifiers
     source          1..7479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        175..478
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        479..682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     633..653
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             711..1418
                     /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(856..874)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1124..1143
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4381..4502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4418..4437)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4472..4491
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4509..4964)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4646..4667)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4858..4877
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4991..5095
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5097..5454
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5097..5288
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5120..5140)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5305..5440
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5489..6283
                     /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             5489..6283
                     /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(5543..5562)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6153..6172
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6471..6490)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6515..6562
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6891..7479
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7380..7399
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
      121 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      181 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      241 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      301 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      361 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      421 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      481 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      541 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      601 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      661 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc atggtgagca
      721 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
      781 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
      841 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
      901 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
      961 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     1021 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
     1081 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
     1141 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
     1201 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     1261 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
     1321 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     1381 gccgccactc caccggcggc atggacgagc tgtacaagtc cggactcaga tctcgagctc
     1441 aagcttcgaa ttctgcagtc gacggtacca tggaaattaa agaggaaggg gcatcagaag
     1501 aagggcagca ctttcttcct acagcccagg ccaatgatcc cggggactgt cagttcacaa
     1561 gtatccagaa gactccaaat gaaccgcagt tggaattcat ccttgcatgc aaggatctcg
     1621 tggctcctgt ccgtgatcgt aaactgaata cactggtgca gatctccgta atccaccccg
     1681 tggagcagag tctgacaaga tactccagca ccgaaattgt ggagggaaca agggacccac
     1741 tgtttttgac tggtgtcaca ttcccatctg agtatcccat ctatgaggag accaaaataa
     1801 aactaacagt ctatgatgtc aaggataagt ctcatgacac cgttcgaacc agtgtcctac
     1861 cagaacataa ggatcccccg ccagaagttg ggcgaagttt cttgggctat gccagtttta
     1921 aagtgggaga gctgctgaag tcaaaggagc aattgctggt cctgagcctg agaacttcag
     1981 atggtggcaa agtggttggc accatagaag tcagtgtcgt gaagatgggg gagattgagg
     2041 atggggaagc cgaccacatc accacagatg tacagggaca aaagtgtgcc ctggtatgtg
     2101 aatgtacagc cccggaaagt gtgagcggaa aagataactt accttttttg aattcagtgt
     2161 taaagaaccc agtatgtaaa ttatatagat ttcccacatc tgacaataag tggatgcgaa
     2221 ttcgagagca gatgtcagag agcattcttt cctttcatat tcctaaggaa ttgatttccc
     2281 ttcacattaa agaagatttg tgcagaaacc aggagataaa agaacttggt gagctttctc
     2341 cacattggga caatctgcga aaaaatgtcc ttactcactg tgatcaaatg gtgaatatgt
     2401 accaagacat tctgacagaa cttagcaagg aaacagggtc ctctttcaaa tcaagcagca
     2461 gcaaaggaga gaaaacatta gaatttgttc caataaatct acatctgcaa agaatgcagg
     2521 tacacagccc tcacttgaaa gatgctctct acgatgtcat cactgtggga gccccagctg
     2581 cccattttca gggatttaag aatggtggtc ttcggaagct actccataga tttgaaacag
     2641 aaagaagaaa taccggatac cagtttattt actattcacc tgaaaacaca gccaaagcaa
     2701 aggaagttct cagcaacatc aatcaactac aacctcttat agcaacccat gcagacctac
     2761 tgcttaattc tgcaagccag cattctccag acagcttgaa gaattcttta aagatgcttt
     2821 cagaaaaaac agagcttttt gtacatgcct tcaaggatca acttgtcagg agtgctcttt
     2881 tagcactcta cactgcaagg ccaggaggca ttcttaagaa gccaccctct cctaagagca
     2941 gcacagagga gagcagtccc caagaccaac ccccagtgat gagagggcag gactccatac
     3001 cacatcattc agactatgat gaggaagagt gggacagggt gtgggccaat gtggggaaga
     3061 gcctgaactg cattattgct atggtggaca aactgattga aagagatggt ggcagtgaag
     3121 gcagtggcgg caacaatgat ggagaaaagg aaccttcatt aacagatgcc attccctctc
     3181 acccaagaga ggactggtat gaacagttgt atcccctcat ccttaccctg aaggactgca
     3241 tgggagaagt ggtgaaccga gccaagcagt ccctgacatt tgtgctcctt caggaacttg
     3301 cgtacagctt gccccagtgt ctgatgctga cgctaagaag agacatcgtc ttcagccaag
     3361 cacttgctgg attggtttgt ggttttatca tcaaattaca gacaagtctg tatgacccag
     3421 gcttcctaca gcagcttcac acagtggggt tgatagtaca atatgaagga ctgctaagta
     3481 catacagcga tgaaattgga atgctagagg acatggccgt tggcatttcc gatttaaaga
     3541 aagttgcatt taaaataatt gaagccaaat ccaatgatgt gttgccagtt ataacaggaa
     3601 gacgagaaca ttacgtggta gaggtcaagc ttccagccag aatgtttgag tcactacctc
     3661 tacagattaa agaaggacag ttgcttcatg tgtatccagt actttttaat gttggaatca
     3721 atgaacagca aactctggct gaaaggtttg gagatgtctc tttgcaagaa agtattaatc
     3781 aggaaaactt cgaacttcta caagaatatt acaagatatt tatggaaaag atgcctcctg
     3841 attatatttc acattttcag gaacaaaatg atttaaaagc attgctagaa aatctccttc
     3901 aaaatatcca atccaaaaaa agaaagaatg tagaaattat gtggctggct gcaacgattt
     3961 gccgcaaact gaatggtatt cgtttcacct gtgctaaaag tgccaaagac aggacatcga
     4021 tgtcagtgac acttgaacaa tgctcaatct tgagagatga gcaccagtta cacaaggact
     4081 tctttatccg agcgctggat tgcatgagaa gagaaggatg ccgcatagag aatgtactga
     4141 agaatatcaa atgcagaaag tatgctttca acatgctaca gctgatggct ttccccaagt
     4201 actacagacc tccagagggg acttatggaa aagctgacac ctaagggccc gggatccacc
     4261 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     4321 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     4381 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4441 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4501 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     4561 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     4621 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     4681 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     4741 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     4801 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     4861 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     4921 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     4981 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5041 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     5101 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5161 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5221 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5281 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5341 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     5401 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     5461 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     5521 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     5581 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     5641 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     5701 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     5761 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     5821 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     5881 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     5941 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     6001 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     6061 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     6121 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     6181 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     6241 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     6301 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     6361 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     6421 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     6481 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     6541 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     6601 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     6661 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     6721 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     6781 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6841 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6901 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6961 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7021 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7081 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7141 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7201 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7261 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7321 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7381 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7441 tttttgtgat gctcgtcagg ggggcggagc ctatggaaa
//
