LOCUS       sequence-343427-        6233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PIK3R1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6233)
  AUTHORS   Zhou Y, Feng Z, Cao F, Liu X, Xia X, Yu CH
  TITLE     Abl-mediated PI3K activation regulates macrophage podosome 
            formation.
  JOURNAL   J Cell Sci. 2020 Jun 8;133(11). pii: jcs.234385. doi: 
            10.1242/jcs.234385.
  PUBMED    32393599
REFERENCE   2  (bases 1 to 6233)
  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: "J Cell 
            Sci."; date: "2020-06-8"; volume: "133(11). pii: jcs.234385. doi"; 
            pages: " 10.1242/jcs.234385"
COMMENT     Sequence Label: mCherry-PIK3R1
FEATURES             Location/Qualifiers
     source          1..6233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        91..1075
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1110..1128
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1110..1127
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1184..1193
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1190..1897
                     /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(1335..1353)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1603..1622
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1797..1820
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    1910..1934
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3297..3321)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3360..3494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3384..3403)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3438..3457
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3607..3627)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3607..3625)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3646..3662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3646..3662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3659..3681)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3670..3686
                     /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(3694..3700,3701..3718,3719..3724))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3739..3760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3877..3894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4048..4636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4128..4147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4807..5598,5599..5667))
                     /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     5430..5449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5668..5772)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5798..6233,1..20))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5935..5956)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6147..6166
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
       61 atcggtgcgg gcctcttcgc tattacgcca gtcgaccgcc aattcaatat ggcgtatatg
      121 gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat ggcgtatatg
      181 gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat ggcggacttg
      241 gcaccatgcc aattcaatat ggcggacctg gcactgtgcc aactggggag gggtctactt
      301 ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc atattgaatt
      361 ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata tataggcaat
      421 atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat gtattggccc
      481 tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc cctcccaatg
      541 ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac tcccccattg
      601 acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg tcctattgac
      661 gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc
      721 cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat tgcccattca
      781 tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact tggcagtaca
      841 tcaatatcta ttaatagtaa cttggcaagt acattactat tggaagtacg ccagggtaca
      901 ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca
      961 tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat
     1021 gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag ggaaccgcca
     1081 ttctgcctgg ggacgtcgga gcaagcttga tttaggtgac actatagaat acaagctact
     1141 tgttcttttt gcaggatccc atcgattcga attcaaggct agcgccacca tggtgagcaa
     1201 gggcgaggag gacaacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     1261 gggctccgtg aacggccgcg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     1321 gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga
     1381 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
     1441 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     1501 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     1561 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     1621 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     1681 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgccg aggtcaagac
     1741 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagct
     1801 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc gcgccgaggg
     1861 ccgccactcc accggcggca tggacgagct gtacaagaga tctcgaatca caagtttgta
     1921 caaaaaagtt ggcatgtaca atactgtttg gaatatggaa gacctggatt tagaatatgc
     1981 caagacagat ataaattgtg gcacagactt gatgttttat atagaaatgg acccaccagc
     2041 actgcctcct aaaccaccaa aacctactac tgtagccaac aacggtatga ataacaatat
     2101 gtccttacaa gatgctgaat ggtactgggg agatatctcg agggaagaag tgaatgaaaa
     2161 acttcgagat acagcagacg ggaccttttt ggtacgagat gcgtctacta aaatgcatgg
     2221 tgattatact cttacactaa ggaaaggggg aaataacaaa ttaatcaaaa tatttcatcg
     2281 agatgggaaa tatggcttct ctgacccatt aaccttcagt tctgtggttg aattaataaa
     2341 ccactaccgg aatgaatctc tagctcagta taatcccaaa ttggatgtga aattacttta
     2401 tccagtatcc aaataccaac aggatcaagt tgtcaaagaa gataatattg aagctgtagg
     2461 gaaaaaatta cataaatata acactcagtt tcaagaaaaa agtcgagaat atgatagatt
     2521 atatgaagaa tatacccgca catcccagga aatccaaatg aaaaggacag ctattgaagc
     2581 atttaatgaa accataaaaa tatttgaaga acagtgccag acccaagagc ggtacagcaa
     2641 agaatacata gaaaagttta aacgtgaagg caatgagaaa gaaatacaaa ggattatgca
     2701 taattatgat aagttgaagt ctcgaatcag tgaaattatt gacagtagaa gaagattgga
     2761 agaagacttg aagaagcagg cagctgagta tcgagaaatt gacaaacgta tgaacagcat
     2821 taaaccagac cttatccagc tgagaaagac gagagaccaa tacttgatgt ggttgactca
     2881 aaaaggtgtt cggcaaaaga agttgaacga gtggttgggc aatgaaaaca ctgaagacca
     2941 atattcactg gtggaagatg atgaagattt gccccatcat gatgagaaga catggaatgt
     3001 tggaagcagc aaccgaaaca aagctgaaaa cctgttgcga gggaagcgag atggcacttt
     3061 tcttgtccgg gagagcagta aacagggctg ctatgcctgc tctgtagtgg tggacggcga
     3121 agtaaagcat tgtgtcataa acaaaacagc aactggctat ggctttgccg agccctataa
     3181 cttgtacagc tctctgaaag aactggtgct acattaccaa cacacctccc ttgtgcagca
     3241 caacgactcc ctcaatgtca cactagccta cccagtatat gcacagcaga ggcgatgccc
     3301 aactttcttg tacaaagtgg tgatcgagcc tctagaacta tagtgagtcg tattacgtag
     3361 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     3421 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     3481 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     3541 tgtgggaggt tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt
     3601 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     3661 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg
     3721 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3781 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3841 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3901 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3961 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4021 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4081 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4141 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4201 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4261 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4321 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4381 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4441 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4501 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4561 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4621 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4681 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4741 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4801 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4861 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4921 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4981 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5041 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5101 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5161 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5221 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5281 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5341 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5401 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5461 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5521 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5581 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5641 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5701 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5761 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     5821 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     5881 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     5941 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     6001 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     6061 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     6121 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     6181 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taa
//
