LOCUS       sequence-337577-        6803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses INPP5E in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6803)
  AUTHORS   Feng Z, Yu CH
  TITLE     PI(3,4)P2-mediated membrane tubulation promotes integrin trafficking
            and invasive cell migration.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 May 11;118(19). pii: 2017645118. doi:
            10.1073/pnas.2017645118.
  PUBMED    33947811
REFERENCE   2  (bases 1 to 6803)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-05-11"; volume: "118(19). pii: 
            2017645118. doi"; pages: " 10.1073/pnas.2017645118"
COMMENT     Sequence Label: mCherry-INPP5E
FEATURES             Location/Qualifiers
     source          1..6803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      109..118
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             115..822
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGREFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(260..278)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     528..547
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     722..745
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    835..859
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2792..2816)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2855..2989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2879..2898)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2933..2952
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3102..3122)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3102..3120)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3141..3157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3141..3157)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3154..3176)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3165..3181
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3189..3195,3196..3213,3214..3219))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3234..3255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3372..3389)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3543..4131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3623..3642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4302..5093,5094..5162))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4925..4944
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5163..5267)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5293..5748)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5430..5451)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5642..5661
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5819..6803
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggctagcgc caccatggtg
      121 agcaagggcg aggaggacaa catggccatc atcaaggagt tcatgcgctt caaggtgcac
      181 atggagggct ccgtgaacgg ccgcgagttc gagatcgagg gcgagggcga gggccgcccc
      241 tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc
      301 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
      361 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
      421 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
      481 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
      541 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
      601 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgccgaggtc
      661 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
      721 aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgcgcc
      781 gagggccgcc actccaccgg cggcatggac gagctgtaca agagatctcg aatcacaagt
      841 ttgtacaaaa aagttggcat gccgtccaag gcggagaatc tgcggccctc cgagccggcc
      901 ccgcagccgc cggaagggag gacgctccaa ggacagcttc ccggcgctcc gccggcccag
      961 cgcgcggggt ccccacccga tgctccgggc tccgagagcc ccgcgcttgc ctgcagcact
     1021 ccggccacgc ccagcggcga ggacccgcca gcccgagcag cacccatcgc cccgcggccc
     1081 cccgccaggc ctcgactgga gcgagccctg tccctggacg acaagggctg gaggaggagg
     1141 cgttttcgag gcagccagga ggacctggaa gcccggaatg ggaccagtcc ctccaggggc
     1201 tcagtgcaga gcgagggccc cggggccccc gcccacagct gctccccgcc ctgcctgagc
     1261 acctccttgc aggaaatccc caagtcccgc ggggtcctga gcagtgagag agggagcccg
     1321 tcctcggggg gtaaccctct ctctggggtg gccagcagct ccccgaacct cccgcacaga
     1381 gacgccgccg tggcggggag ctcgcccagg ctgcccagcc tgctgccccc gcgcccaccg
     1441 cctgccctga gcctggacat cgcctccgac tccctgagga cagcaaacaa ggtcgactcg
     1501 gatcttgcag actacaagct ccgcgcgcag ccgctcctgg tgcgggccca cagcagcctg
     1561 ggccccggcc ggccgcggag ccccctggcc tgcgacgact gttcccttcg ctcggccaaa
     1621 tcctccttca gcctcctggc gcccatccgc agcaaggacg tccgcagcag gagttacctg
     1681 gagggcagcc tcctggccag cggggccctg ttgggggcgg atgagctggc ccgctacttc
     1741 ccagaccgga acgtggcact cttcgtggcc acctggaaca tgcagggcca gaaggagctc
     1801 ccgcccagcc tggacgagtt cctgctccca gccgaggccg actatgccca ggacctgtat
     1861 gtcatcgggg tccaggaggg ctgttctgac aggcgggagt gggagactcg tctgcaggag
     1921 acgctgggcc cgcactatgt gctgctgtcc tcggcggccc acggcgtgct ctacatgtcg
     1981 ctcttcatcc gcagggacct catctggttc tgctcagagg tggagtgctc cacggtgacc
     2041 acacgcatcg tgtctcagat caagaccaag ggggccttgg gcatcagctt cacctttttt
     2101 ggcacttcct tcctcttcat cacgtcccac ttcacctcag gtgacgggaa ggtggcggag
     2161 cggctgctgg actacaccag gactgtacaa gccctggtcc tgcccagaaa tgtgcccgac
     2221 accaacccct atcgctccag cgcagcggac gtcaccaccc gcttcgatga ggtgttctgg
     2281 tttggagact tcaacttccg cctgagtggc gggcgcacag tcgtggacgc cctcctgtgc
     2341 cagggcctgg tggtggacgt gccggcgctg ctgcagcacg accagctcat ccgggagatg
     2401 cggaaagggt ccatcttcaa gggcttccag gagccggaca tccacttcct cccatcatac
     2461 aagtttgaca tcgggaagga cacgtacgac agcacctcca agcagaggac gccctcatac
     2521 acggaccgcg tcttgtacag aagccgccac aagggtgaca tctgtcctgt gagctactct
     2581 tcctgccccg ggatcaagac gtccgaccac cgccctgtgt atggcctctt ccgggtgaaa
     2641 gtgaggccgg ggcgagacaa cattccgttg gcagctggca aatttgatag agaactgtac
     2701 ttactaggaa ttaaaagacg gatttcgaag gagattcaga ggcagcaagc actacagagt
     2761 cagaactcca gcaccatctg ctccgtttct tgcccaactt tcttgtacaa agtggtgatc
     2821 gagcctctag aactatagtg agtcgtatta cgtagatcca gacatgataa gatacattga
     2881 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2941 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3001 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc
     3061 cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg agggttaatt
     3121 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3181 attccacaca acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg
     3241 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3301 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3361 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3421 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3481 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3541 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3601 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3661 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3721 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3781 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3841 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3901 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3961 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4021 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4081 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4141 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4201 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4261 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4321 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4381 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4441 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4501 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4561 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4621 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4681 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4741 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4801 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4861 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4921 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4981 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5041 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5101 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5161 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5221 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5281 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     5341 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     5401 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     5461 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     5521 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     5581 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5641 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     5701 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
     5761 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt
     5821 cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg tggatctgga
     5881 cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga
     5941 ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg cggacctggc
     6001 actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga ggggtcttgg
     6061 ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg ccatattggc
     6121 tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa tatggctatt
     6181 ggccaggttc aatactatgt attggcccta tgccatatag tattccatat atgggttttc
     6241 ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct
     6301 aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta
     6361 ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg acgtcaatta
     6421 cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc caccattgac
     6481 gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat tgacgtcaat
     6541 gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact tggcaagtac
     6601 attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt
     6661 aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga
     6721 cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata
     6781 taagcaatgc tcgtttaggg aac
//
