LOCUS       sequence-337580-        7619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses INPP5B in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7619)
  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 7619)
  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-INPP5B
FEATURES             Location/Qualifiers
     source          1..7619
                     /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(3608..3632)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3671..3805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3695..3714)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3749..3768
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3918..3938)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3918..3936)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3957..3973)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3957..3973)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3970..3992)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3981..3997
                     /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(4005..4011,4012..4029,4030..4035))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4050..4071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4188..4205)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4359..4947)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4439..4458)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5118..5909,5910..5978))
                     /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     5741..5760
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5979..6083)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6109..6564)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6246..6267)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6458..6477
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6635..7619
                     /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 aagcaggctc caccatggac cagtctgtgg caatccagga gacgctggct
      901 gagggggaat actgcgtcat cgcggtgcaa ggtgtgctgt gtgaggggga cagccggcag
      961 agccgcctcc tgggactcgt gcgctaccgc ctggagcacg gcggccagga acacgctctc
     1021 ttcctctata cgcaccggag gatggccatt accggggacg atgtctctct ggaccagata
     1081 gtgccagtct cgcgggattt tacgctggaa gaagtgtccc cagatggtga actctacatc
     1141 cttggctcag atgtgaccgt ccagctggac acagcagagc ttagcctcgt attccaactg
     1201 ccctttggtt cacaaaccag gatgttcctc cacgaagttg ccagggcctg tccaggcttc
     1261 gattctgcga cccgggatcc tgaattcctg tggctgtctc ggtataggtg cgcagagctg
     1321 gagctggaga tgccaacgcc gcgcggttgt aactcggccc tagttacctg gccagggtac
     1381 gcgacaattg gcggaggtgg ttctaacttt gatggtttgc gaccaaatgg gaagggagtg
     1441 cctatggacc aaagctccag gggtcaagat aaaccagaaa gcttgcaacc aagacagaat
     1501 aaatccaagt ccgaaattac tgacatggtt cgctcctcca ctatcacagt gtcggacaag
     1561 gctcatattt tatccatgca gaagtttgga ctgcgagata caattgtgaa atcacatcta
     1621 ctacagaaag aagaggatta cacctatatc cagaacttca ggttttttgc gggaacatac
     1681 aatgtaaatg ggcagtcccc caaagaatgc ctccggctgt ggctgagcaa tggtatccag
     1741 gcaccagatg tctattgtgt agggttccag gagcttgatc tgagtaagga agcttttttc
     1801 tttcacgata ccccaaagga ggaagagtgg ttcaaagctg tgtcagaggg tcttcatcca
     1861 gatgccaaat atgcaaaggt gaagcttatc cgactggttg ggattatgct gctgttatat
     1921 gtcaaacagg agcatgcagc ttatatctca gaagtggaag ccgagactgt ggggacagga
     1981 atcatgggga ggatgggcaa caagggaggc gtggcgatca ggttccagtt ccacaacacc
     2041 agcatctgcg ttgtgaattc tcacttggca gcccacattg aagagtatga gaggaggaac
     2101 caggactata aggacatttg ttctcgaatg cagttttgtc agcctgaccc aagccttccc
     2161 cctctcacca tcagcaacca tgatgtgatc ttgtggctgg gggacctcaa ctacaggata
     2221 gaagagctgg atgtggaaaa agtgaaaaag ctcatcgaag agaaggactt tcaaatgctg
     2281 tatgcatatg atcagctgaa aattcaggtg gccgcaaaga ctgtctttga aggcttcaca
     2341 gagggtgagc tcacattcca gcctacttac aagtatgata cgggctctga cgactgggat
     2401 accagtgaga agtgccgtgc tcctgcctgg tgtgatcgga ttctctggaa agggaagaac
     2461 atcactcagc tgagttacca gagccacatg gccctgaaga ccagtgacca caagcctgtc
     2521 agctcagtgt ttgacatcgg ggtgagggtc gtaaatgacg agctttaccg gaagacactg
     2581 gaggaaattg ttcgatccct ggataagatg gaaaatgcca acattccttc tgtgtccctg
     2641 tccaagcgag agttctgttt tcagaatgtg aagtacatgc aattgaaagt agaatccttt
     2701 acaattcata atggacaagt accctgtcat tttgaattca tcaacaagcc tgatgaagag
     2761 tcttactgta agcagtggct gaatgccaac cccagcagag gcttcctcct gccagattct
     2821 gatgttgaga ttgacttgga gctcttcgta aataagatga cagctacaaa gctcaactcg
     2881 ggtgaagaca aaattgagga cattctggtt ctgcacttgg acaggggaaa ggattacttt
     2941 ttgtctgtgt ctgggaacta cctgcccagc tgttttggtt ctcccattca tacactgtgt
     3001 tacatgagag agccaatctt ggacctacca cttgaaacca ttagtgagct gactctgatg
     3061 ccagtatgga ctggagatga tgggagccag ttggatagcc ccatggaaat ccccaaagag
     3121 ctctggatga tggttgatta cctgtaccga aatgctgtcc agcaggaaga tctgtttcag
     3181 caaccaggcc tgaggtcaga atttgaacat atcagggact gcttggatac tggaatgatt
     3241 gataacctct ctgccagcaa tcattctgta gccgaagcct tgctgctttt cctggagagc
     3301 cttccagagc ctgtcatctg ttacagcacc taccataact gcttggagtg ttctggcaac
     3361 tacacagcaa gcaaacaggt catttctact ctccccatat tccacaaaaa tgtcttccac
     3421 tacttgatgg cgtttttgcg agaactgctg aaaaattcag caaaaaatca tttggatgag
     3481 aatattctag ctagcatatt tggcagctta ttgcttcgaa acccagctgg tcaccaaaag
     3541 cttgatatga cagagaagaa gaaggctcaa gaatttattc accagttcct ctgcaaccca
     3601 ctcttggacc cagctttctt gtacaaagtg gtgatcgagc ctctagaact atagtgagtc
     3661 gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3721 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3781 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3841 tcagggggag gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc attgggcccg
     3901 gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     3961 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccggg
     4021 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4081 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4141 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4201 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4261 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4321 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4381 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4441 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4501 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4561 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4621 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4681 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4741 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4801 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4861 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4921 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4981 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5041 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5101 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5161 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5221 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5281 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5341 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5401 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5461 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5521 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5581 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5641 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5701 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5761 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5821 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5881 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5941 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6001 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6061 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     6121 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     6181 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     6241 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     6301 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     6361 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     6421 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6481 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6541 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     6601 ggcgatcggt gcgggcctct tcgctattac gccagtcgac cgccaattca atatggcgta
     6661 tatggactca tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta
     6721 tatggactcg tgccaattca atatggtgga tctggacccc agccaattca atatggcgga
     6781 cttggcacca tgccaattca atatggcgga cctggcactg tgccaactgg ggaggggtct
     6841 acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg
     6901 aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg
     6961 caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg
     7021 gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc
     7081 aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc
     7141 attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat
     7201 tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa
     7261 tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca
     7321 ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag
     7381 tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag tacgccaggg
     7441 tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag
     7501 tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt
     7561 aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaac
//
