LOCUS       sequence-343420-        6261 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 6261)
  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 6261)
  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: EGFP-PIK3R1
FEATURES             Location/Qualifiers
     source          1..6261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        88..1072
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1107..1125
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1107..1124
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1191..1200
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1197..1199,1200..1202,1203..1913)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1197..1199,1200..1202,1203..1913)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1242..1263)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1503..1522)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1850..1871
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1935..1959
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3322..3346)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3385..3519
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3409..3428)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3463..3482
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3632..3652)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3632..3650)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3671..3687)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3671..3687)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3684..3706)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3695..3711
                     /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(3719..3725,3726..3743,3744..3749))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3764..3785
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3902..3919)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4073..4661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4153..4172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4832..5623,5624..5692))
                     /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     5455..5474
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5693..5797)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5823..6261,1..17))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5960..5981)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6172..6191
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
       61 ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat tcaatatggc gtatatggac
      121 tcatgccaat tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac
      181 tcgtgccaat tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca
      241 ccatgccaat tcaatatggc ggacctggca ctgtgccaac tggggagggg tctacttggc
      301 acggtgccaa gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc
      361 atggtgccaa taatggcggc catattggct atatgccagg atcaatatat aggcaatatc
      421 caatatggcc ctatgccaat atggctattg gccaggttca atactatgta ttggccctat
      481 gccatatagt attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc
      541 ggtcccatat accatatatg gggcttccta ataccgccca tagccactcc cccattgacg
      601 tcaatggtct ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta
      661 tatggcgcct cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat
      721 tgacgtcaat aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat
      781 ccgttctcac gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca
      841 atatctatta atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg
      901 gcagtactcc cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc
      961 ccattgacgt caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg
     1021 cggtaggcgt gcctaatggg aggtctatat aagcaatgct cgtttaggga accgccattc
     1081 tgcctgggga cgtcggagca agcttgattt aggtgacact atagaataca agctacttgt
     1141 tctttttgca ggatcccatc gattcgaatt caaggctagc gctaccggtc gccaccatgg
     1201 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1261 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1321 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1381 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1441 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1501 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1561 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1621 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1681 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1741 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1801 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1861 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac
     1921 tcagatctcg aatcacaagt ttgtacaaaa aagttggcat gtacaatact gtttggaata
     1981 tggaagacct ggatttagaa tatgccaaga cagatataaa ttgtggcaca gacttgatgt
     2041 tttatataga aatggaccca ccagcactgc ctcctaaacc accaaaacct actactgtag
     2101 ccaacaacgg tatgaataac aatatgtcct tacaagatgc tgaatggtac tggggagata
     2161 tctcgaggga agaagtgaat gaaaaacttc gagatacagc agacgggacc tttttggtac
     2221 gagatgcgtc tactaaaatg catggtgatt atactcttac actaaggaaa gggggaaata
     2281 acaaattaat caaaatattt catcgagatg ggaaatatgg cttctctgac ccattaacct
     2341 tcagttctgt ggttgaatta ataaaccact accggaatga atctctagct cagtataatc
     2401 ccaaattgga tgtgaaatta ctttatccag tatccaaata ccaacaggat caagttgtca
     2461 aagaagataa tattgaagct gtagggaaaa aattacataa atataacact cagtttcaag
     2521 aaaaaagtcg agaatatgat agattatatg aagaatatac ccgcacatcc caggaaatcc
     2581 aaatgaaaag gacagctatt gaagcattta atgaaaccat aaaaatattt gaagaacagt
     2641 gccagaccca agagcggtac agcaaagaat acatagaaaa gtttaaacgt gaaggcaatg
     2701 agaaagaaat acaaaggatt atgcataatt atgataagtt gaagtctcga atcagtgaaa
     2761 ttattgacag tagaagaaga ttggaagaag acttgaagaa gcaggcagct gagtatcgag
     2821 aaattgacaa acgtatgaac agcattaaac cagaccttat ccagctgaga aagacgagag
     2881 accaatactt gatgtggttg actcaaaaag gtgttcggca aaagaagttg aacgagtggt
     2941 tgggcaatga aaacactgaa gaccaatatt cactggtgga agatgatgaa gatttgcccc
     3001 atcatgatga gaagacatgg aatgttggaa gcagcaaccg aaacaaagct gaaaacctgt
     3061 tgcgagggaa gcgagatggc acttttcttg tccgggagag cagtaaacag ggctgctatg
     3121 cctgctctgt agtggtggac ggcgaagtaa agcattgtgt cataaacaaa acagcaactg
     3181 gctatggctt tgccgagccc tataacttgt acagctctct gaaagaactg gtgctacatt
     3241 accaacacac ctcccttgtg cagcacaacg actccctcaa tgtcacacta gcctacccag
     3301 tatatgcaca gcagaggcga tgcccaactt tcttgtacaa agtggtgatc gagcctctag
     3361 aactatagtg agtcgtatta cgtagatcca gacatgataa gatacattga tgagtttgga
     3421 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3481 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3541 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca
     3601 atgcattggg cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     3661 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3721 acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3781 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3841 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3901 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3961 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4021 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4081 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4141 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4201 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4261 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4321 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4381 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4441 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4501 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4561 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4621 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4681 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4741 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4801 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4861 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4921 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4981 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5041 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5101 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5161 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5221 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5281 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5341 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5401 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5461 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5521 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5581 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5641 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5701 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5761 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5821 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     5881 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     5941 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     6001 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     6061 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     6121 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     6181 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     6241 cacacccgcc gcgcttaatg c
//
