LOCUS       sequence-337576-        6791 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses INPP5J in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6791)
  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 6791)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-INPP5J
FEATURES             Location/Qualifiers
     source          1..6791
                     /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(2780..2804)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2843..2977
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2867..2886)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2921..2940
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3090..3110)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3090..3108)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3129..3145)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3129..3145)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3142..3164)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3153..3169
                     /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(3177..3183,3184..3201,3202..3207))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3222..3243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3360..3377)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3531..4119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3611..3630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4290..5081,5082..5150))
                     /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     4913..4932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5151..5255)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5281..5736)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5418..5439)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5630..5649
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5807..6791
                     /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 aagttggcac catggccctc ccaaggcttg gcacccagag tacagggcct
      901 ggcaggtgcc tgagccccaa ccttcaggcc caagaagccc cagccccagt caccacctcc
      961 tcttctacat ccaccctgtc atcctcccct tggtcagctc agcctacctg gaagagcgac
     1021 cccggcttcc ggatcactgt ggtcacatgg aacgtgggca ctgccatgcc cccagacgat
     1081 gtcacatccc tcctccacct gggcggtggt gacgacagcg acggcgcaga catgatcgcc
     1141 atagggttgc aggaagtgaa ctccatgctc aacaagcgac tcaaggacgc cctcttcacg
     1201 gaccagtgga gtgagctgtt catggatgcg ctagggccct tcaacttcgt gctggtgagt
     1261 tcggtgagga tgcagggtgt catcctgctg ctgttcgcca agtactacca cctgcccttc
     1321 ctgcgagacg tgcagaccga ctgcacgcgc actggcctgg gcggctactg gggtaacaag
     1381 ggtggcgtga gcgtgcgcct ggcggccttc gggcacatgc tctgcttcct gaactgccac
     1441 ttgcctgcgc atatggacaa ggcggagcag cgcaaagaca acttccagac catcctcagc
     1501 ctccagcagt tccaagggcc gggcgcacag ggcatcctgg atcatgacct cgtgttctgg
     1561 ttcggggacc tgaacttccg cattgagagc tatgacctgc actttgtcaa gtttgccatc
     1621 gacagtgacc agctccatca gctctgggag aaggaccagc tcaacatggc caagaacacc
     1681 tggcccattc tgaagggctt tcaggagggg cccctcaact tcgctcccac cttcaagttt
     1741 gatgtgggta ccaacaaata cgataccagt gccaagaaac ggaagccagc ttggacagac
     1801 cgtatcctat ggaaggtcaa ggctccaggt gggggtccca gcccctcagg acggaagagc
     1861 caccgactcc aggtgacgca gcacagctac cgcagccaca tggaatacac agtcagcgac
     1921 cacaagcctg tggctgccca gttcctcctg cagtttgcct tcagggacga catgccactg
     1981 gtgcggctgg aggtggcaga tgagtgggtg cggcccgagc aggcggtggt gaggtaccgc
     2041 atggaaacag tgttcgcccg cagctcctgg gactggatcg gcttataccg ggtgggtttc
     2101 cgccattgca aggactatgt ggcttatgtc cgggccaaac atgaagatgt ggatgggaat
     2161 acataccagg taacattcag tgaggaatca ctgcccaagg gccatggaga cttcatcctg
     2221 ggctactata gtcacaacca cagcatcctc atcggcatca ctgaaccctt ccagatctcg
     2281 ctgccttcct cggagttggc cagcagcagc acagacagct caggcaccag ctcagaggga
     2341 gaggatgaca gcacactgga gctccttgca cccaagtccc gcagccccag tcctggcaag
     2401 tccaagcgac accgcagccg cagcccggga ctggccaggt tccctgggct tgccctacgg
     2461 ccctcatccc gtgaacgccg tggtgccagc cgtagcccct caccccagag ccgccgcctg
     2521 tcccgagtgg ctcctgacag gagcagtaat ggcagcagcc ggggcagtag tgaagagggg
     2581 ccctctgggt tgcctggccc ctgggccttc ccaccagctg tgcctcgaag cctgggcctg
     2641 ttgcccgcct tgcgcctaga gactgtagac cctggtggtg gtggctcctg gggacctgat
     2701 cgggaggccc tggcgcccaa cagcctgtct cctagtcccc agggccatcg ggggctggag
     2761 gaagggggcc tggggccctt gccaactttc ttgtacaaag tggtgatcga gcctctagaa
     2821 ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca
     2881 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2941 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3001 tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg cggcgccaat
     3061 gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     3121 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3181 atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3241 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3301 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3361 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3421 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3481 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3541 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3601 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3661 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3721 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3781 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3841 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3901 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3961 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4021 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4081 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4141 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4201 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4261 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4321 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4381 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4441 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4501 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4561 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4621 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4681 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4741 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4801 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4861 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4921 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4981 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5041 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5101 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5161 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5221 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5281 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     5341 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     5401 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     5461 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     5521 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     5581 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     5641 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     5701 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     5761 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg accgccaatt
     5821 caatatggcg tatatggact catgccaatt caatatggtg gatctggacc tgtgccaatt
     5881 caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc ccagccaatt
     5941 caatatggcg gacttggcac catgccaatt caatatggcg gacctggcac tgtgccaact
     6001 ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag
     6061 tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta tatgccagga
     6121 tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa
     6181 tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag
     6241 atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat
     6301 agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat
     6361 gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc
     6421 cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg
     6481 gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg
     6541 cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga
     6601 agtacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg
     6661 atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg
     6721 ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc
     6781 gtttagggaa c
//
