LOCUS       sequence-349799-        8066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PIK3CG in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8066)
  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 8066)
  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-PIK3CG
FEATURES             Location/Qualifiers
     source          1..8066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             32..826
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             32..826
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(86..105)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     696..715
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1014..1033)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1058..1105
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1434..2022
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1923..1942
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2197..2500
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2501..2704
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2655..2675
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2743..2752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2749..2751,2752..2754,2755..3465)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2749..2751,2752..2754,2755..3465)
                     /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(2794..2815)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3055..3074)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3402..3423
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6420..6431
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    6990..7111
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7027..7046)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7081..7100
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(7118..7573)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7255..7276)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7467..7486
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7600..7704
                     /gene="bla"
                     /label=AmpR promoter
     promoter        7706..8063
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7706..7897
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7729..7749)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7914..8049
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
       61 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
      121 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
      181 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
      241 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
      301 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
      361 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
      421 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
      481 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
      541 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
      601 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
      661 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
      721 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
      781 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
      841 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
      901 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
      961 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     1021 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     1081 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     1141 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     1201 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     1261 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     1321 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     1381 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1441 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1501 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1561 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     1621 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1681 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1741 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1801 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1861 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1921 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1981 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2041 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2101 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     2161 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     2221 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     2281 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     2341 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     2401 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     2461 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     2521 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     2581 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     2641 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     2701 agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggtgagcaag
     2761 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2821 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     2881 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2941 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     3001 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     3061 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     3121 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     3181 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     3241 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     3301 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     3361 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     3421 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct
     3481 cgagctcaag cttcgaattc tgcagtcgac accatggagc tggagaacta taaacagccc
     3541 gtggtgctga gagaggacaa ctgccgaagg cgccggagga tgaagccgcg cagtgctgcg
     3601 gccagcctgt cctccatgga gctcatcccc atcgagttcg tgctgcccac cagccagcgc
     3661 aaatgcaaga gccccgaaac ggcgctgctg cacgtggccg gccacggcaa cgtggagcag
     3721 atgaaggccc aggtgtggct gcgagcgctg gagaccagcg tggcggcgga cttctaccac
     3781 cggctgggac cgcatcactt cctcctgctc tatcagaaga aggggcagtg gtacgagatc
     3841 tacgacaagt accaggtggt gcagactctg gactgcctgc gctactggaa ggccacgcac
     3901 cggagcccgg gccagatcca cctggtgcag cggcacccgc cctccgagga gtcccaagcc
     3961 ttccagcggc agctcacggc gctgattggc tatgacgtca ctgacgtcag caacgtgcac
     4021 gacgatgagc tggagttcac gcgccgtggc ttggtgaccc cgcgcatggc ggaggtggcc
     4081 agccgcgacc ccaagctcta cgccatgcac ccgtgggtga cgtccaagcc cctcccggag
     4141 tacctgtgga agaagattgc caacaactgc atcttcatcg tcattcaccg cagcaccacc
     4201 agccagacca ttaaggtctc acccgacgac acccccggcg ccatcctgca gagcttcttc
     4261 accaagatgg ccaagaagaa atctctgatg gatattcccg aaagccaaag cgaacaggat
     4321 tttgtgctgc gcgtctgtgg ccgggatgag tacctggtgg gcgaaacgcc catcaaaaac
     4381 ttccagtggg tgaggcactg cctcaagaac ggagaagaga ttcacgtggt actggacacg
     4441 cctccagacc cggccctaga cgaggtgagg aaggaagagt ggccgctggt ggacgactgc
     4501 acgggagtca ccggctacca tgagcagctt accatccacg gcaaggacca cgagagtgtg
     4561 ttcaccgtgt ccctgtggga ctgcgaccgc aagttcaggg tcaagatcag aggcattgat
     4621 atccccgtcc tgcctcggaa caccgacctc acagtttttg tagaggcaaa catccagcat
     4681 gggcaacaag tcctttgcca aaggagaacc agccccaaac ccttcacaga ggaggtgctg
     4741 tggaatgtgt ggcttgagtt cagtatcaaa atcaaagact tgcccaaagg ggctctactg
     4801 aacctccaga tctactgcgg taaagctcca gcactgtcca gcaaggcctc tgcagagtcc
     4861 cccagttctg agtccaaggg caaagttcag cttctctatt atgtgaacct gctgctgata
     4921 gaccaccgtt tcctcctgcg ccgtggagaa tacgtcctcc acatgtggca gatatctggg
     4981 aagggagaag accaaggaag cttcaatgct gacaaactca cgtctgcaac taacccagac
     5041 aaggagaact caatgtccat ctccattctt ctggacaatt actgccaccc gatagccctg
     5101 cctaagcatc agcccacccc tgacccggaa ggggaccggg ttcgagcaga aatgcccaac
     5161 cagcttcgca agcaattgga ggcgatcata gccactgatc cacttaaccc tctcacagca
     5221 gaggacaaag aattgctctg gcattttaga tacgaaagcc ttaagcaccc aaaagcatat
     5281 cctaagctat ttagttcagt gaaatgggga cagcaagaaa ttgtggccaa aacataccaa
     5341 ttgttggcca gaagggaagt ctgggatcaa agtgctttgg atgttgggtt aacaatgcag
     5401 ctcctggact gcaacttctc agatgaaaat gtaagagcca ttgcagttca gaaactggag
     5461 agcttggagg acgatgatgt tctgcattac cttctacaat tggtccaggc tgtgaaattt
     5521 gaaccatacc atgatagtgc ccttgccaga tttctgctga agcgtggttt aagaaacaaa
     5581 agaattggtc actttttgtt ttggttcttg agaagtgaga tagcccagtc cagacactat
     5641 cagcagaggt tcgctgtgat tctggaagcc tatctgaggg gctgtggcac agccatgctg
     5701 cacgacttta cccaacaagt ccaagtaatc gagatgttac aaaaagtcac ccttgatatt
     5761 aaatcgctct ctgctgaaaa gtatgacgtc agttcccaag ttatttcaca acttaaacaa
     5821 aagcttgaaa acctgcagaa ttctcaactc cccgaaagct ttagagttcc atatgatcct
     5881 ggactgaaag caggagcgct ggcaattgaa aaatgtaaag taatggcctc caagaaaaaa
     5941 ccactatggc ttgagtttaa atgtgccgat cctacagccc tatcaaatga aacaattgga
     6001 attatcttta aacatggtga tgatctgcgc caagacatgc ttattttaca gattctacga
     6061 atcatggagt ctatttggga gactgaatct ttggatctat gcctcctgcc atatggttgc
     6121 atttcaactg gtgacaaaat aggaatgatc gagattgtga aagacgccac gacaattgcc
     6181 aaaattcagc aaagcacagt gggcaacacg ggagcattta aagatgaagt cctgaatcac
     6241 tggctcaaag aaaaatcccc tactgaagaa aagtttcagg cagcagtgga gagatttgtt
     6301 tattcctgtg caggctactg tgtggcaacc tttgttcttg gaataggcga cagacacaat
     6361 gacaatatta tgatcaccga gacaggaaac ctatttcata ttgacttcgg gcacattctt
     6421 gggaattaca aaagtttcct gggcattaat aaagagagag tgccatttgt gctaacccct
     6481 gacttcctct ttgtgatggg aacttctgga aagaagacaa gcccacactt ccagaaattt
     6541 caggacatct gtgttaaggc ttatctagcc cttcgtcatc acacaaacct actgatcatc
     6601 ctgttctcca tgatgctgat gacaggaatg ccccagttaa caagcaaaga agacattgaa
     6661 tatatccggg atgccctcac agtggggaaa aatgaggagg atgctaaaaa gtattttctt
     6721 gatcagatcg aagtttgcag agacaaagga tggactgtgc agtttaattg gtttctacat
     6781 cttgttcttg gcatcaaaca aggagagaaa cattcagcct tgggaagctt tctagaccat
     6841 tcgtttggcg cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata
     6901 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     6961 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     7021 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     7081 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     7141 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     7201 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     7261 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     7321 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     7381 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     7441 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     7501 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     7561 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     7621 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     7681 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     7741 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     7801 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     7861 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     7921 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     7981 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     8041 ttggaggcct aggcttttgc aaagat
//
