LOCUS       sequence-337698-        9062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PIK3C2G in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9062)
  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 9062)
  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: EGFP-PIK3C2G
FEATURES             Location/Qualifiers
     source          1..9062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5850..5971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5887..5906)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5941..5960
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5978..6433)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6115..6136)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6327..6346
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6460..6564
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6566..6923
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6566..6757
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6589..6609)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6774..6909
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6958..7752
                     /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             6958..7752
                     /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(7012..7031)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7622..7641
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7940..7959)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7984..8031
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8360..8948
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8849..8868
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacatggca
     1381 tattcttggc aaacggatcc aaatcctaat gaatcacacg aaaagcagta tgaacaccaa
     1441 gaatttctct ttgtaaatca accccattct tctagccaag tcagtctggg ttttgatcag
     1501 atagtagatg agatcagtgg caaaattcca cactacgaga gtgaaattga tgaaaacacc
     1561 ttttttgtgc ccactgcacc aaaatgggac tcaacagggc attcattaaa tgaagcacac
     1621 caaatatcct tgaatgaatt cacttctaaa agccgtgaac tctcctggca tcaagttagc
     1681 aaagcaccag caattggttt tagtccttct gtgttaccaa aacctcaaaa tacgaataaa
     1741 gaatgctcct ggggaagcat aggaaaacat catggtgctg atgattccag attcagtatt
     1801 ttagctctat cattcacaag tttggataaa attaatctag agaaagaatt agaaaatgaa
     1861 aatcataact accatatagg atttgaaagt agcattcctc caacaaattc atccttctca
     1921 agtgacttca tgccgaaaga agagaataaa aggagtggac atgtgaacat tgtggaacca
     1981 tctttgatgc ttttgaaagg ctctcttcaa cccggaatgt gggaaagtac atggcagaag
     2041 aatatagagt caataggttg ttccattcag ctagtggaag tacctcaaag cagcaatacg
     2101 agtctggcct ctttttgcaa caaagtaaaa aaaatcagag aaagatatca tgcagctgat
     2161 gttaatttca attctgggaa gatctggagc actactacag catttccgta tcagctcttt
     2221 tctaagacca agtttaatat acatattttt attgataact caacacaacc tcttcatttt
     2281 atgccatgtg ctaattatct tgtcaaagat ctaattgcag aaattctgca tttttgcaca
     2341 aatgaccagc tactccccaa agatcatatt ctaagtgtat gtggctctga agaattttta
     2401 caaaacgacc actgtttggg gagccacaaa atgtttcaaa aagataaatc tgttattcag
     2461 ctccacctgc agaaaagtag ggaagctcca ggaaagctat ctcgaaagca tgaagaggac
     2521 cacagtcagt tttatctgaa tcaacttcta gaatttatgc atatttggaa agtatccaga
     2581 caatgtctct taacactcat cagaaaatat gacttccacc tgaaatacct attgaaaacc
     2641 caggaaaacg tgtataatat tattgaagaa gttaaaaaaa tatgcagtgt tctagggtgt
     2701 gtggaaacca aacaaattac agatgcagta aatgaactaa gtctaattct tcagagaaaa
     2761 ggagagaatt tttatcaaag ttcagagact tcagcaaaag gcttgataga gaaggtaaca
     2821 actgaactat ccacatccat ctaccagcta atcaatgtct actgtaacag cttttatgca
     2881 gattttcagc ctgtaaatgt acctagatgc acttcctatc taaatcccgg gcttccttcc
     2941 cacctcagct tcacagtgta tgcagcacac aacattccag aaacctgggt gcacaggatc
     3001 aattttcccc ttgaaataaa gtcacttcca agggaatcca tgctcactgt aaaactgttt
     3061 gggattgcct gtgcaaccaa caatgcaaat ttactggcgt ggacttgtct tccactgttt
     3121 ccaaaagaaa aatccattct cgggtctatg ctgttcagca tgacattaca gagtgagcct
     3181 cccgtagaaa tgataactcc aggagtgtgg gatgtaagtc agccatcccc ggtgaccctg
     3241 cagattgatt ttccagctac tgggtgggag tatatgaaac ctgattctga agagaataga
     3301 agtaatcttg aagagccact aaaggagtgt ataaaacata ttgccagact ttcacagaaa
     3361 cagactcccc tactactctc tgaagaaaag aaaagatatt tatggtttta tcgcttctac
     3421 tgcaataatg aaaactgctc ccttccttta gtcctgggta gtgcccctgg atgggatgaa
     3481 aggactgttt cagaaatgca taccattttg agaagatgga cattttctca acctttagag
     3541 gctcttgggc ttttgacttc cagttttcca gatcaagaaa ttcgtaaagt ggcagttcaa
     3601 caattagaca acctcttgaa tgatgaacta ctggaatatc tcccacagct agttcaggct
     3661 gtcaagtttg aatggaacct tgagagtcct ttagtgcaac ttctactcca ccgctccttg
     3721 cagagcatcc aggttgccca tcgtctttac tggctgctaa aaaatgcaga aaatgaagct
     3781 tattttaaaa gctggtatca gaagctacta gctgctctcc aattctgtgc aggtaaagcc
     3841 ttgaatgatg agttttccaa ggagcagaaa cttatcaaaa ttctgggaga tattggggaa
     3901 agagtcaagt ctgccagtga ccatcaaaga caggaggtac tgaagaaaga aattggcaga
     3961 ctagaagagt tctttcaaga tgtaaatact tgtcatcttc ctctgaaccc tgccctatgt
     4021 ataaaaggga ttgatcacga tgcatgttca tattttacat ctaatgcttt gccattgaag
     4081 attactttca tcaatgctaa tctgatgggc aaaaacatca gcattatttt taaggctgga
     4141 gatgatcttc gtcaggatat gcttgttctg cagcttattc aagtgatgga caatatttgg
     4201 ctgcaggaag gcttggatat gcaaatgatc atttatagat gtctatccac aggaaaagac
     4261 caaggattgg tgcagatggt acctgatgct gtgaccctag caaagattca tcgccattct
     4321 ggactgatag gaccattgaa agaaaataca attaaaaagt ggttcagtca gcacaaccac
     4381 ttaaaggcag attatgaaaa ggccttgagg aactttttct actcctgtgc tggctggtgt
     4441 gtggtaacat tcatcctggg agtatgtgac cgtcacaatg ataatatcat gctgacaaag
     4501 tcgggccaca tgtttcatat tgactttgga aaattcttag gtcatgcaca aacatttgga
     4561 gggataaaaa gggaccgagc tcctttcatt tttacttcag agatggaata ctttattaca
     4621 gagggtggga aaaacccaca gcattttcaa gattttgtgg aactttgctg tcgtgcttat
     4681 aatattatca gaaagcacag ccaactgctc ttgaacctgc tggaaatgat gctgtatgca
     4741 ggactgcctg agctaagtgg aattcaagac ctgaaatatg tgtataataa tcttcgtcca
     4801 caagacacag acctggaagc aacaagtcat tttaccaaga aaataaagga aagtctggag
     4861 tgtttccctg ttaaattgaa taacttgatc cacacacttg cacaaatgtc agccataagc
     4921 cctgccaaat ctacttcaca gacttttcct caggaatcct gtttgctgag tacaactagg
     4981 tcgattgaaa gagcaacaat tttagggttc agcaagaaat ccagtaatct gtatctgatc
     5041 caggtgacac acagcaacaa cgaaacaagc ctgacagaaa aatcatttga gcagttttca
     5101 aaacttcaca gccaacttca gaagcagttt gcatcactga ctctcccaga gtttcctcat
     5161 tggtggcacc taccttttac aaattcagat cacagaagat tcagagatct aaatcattac
     5221 atggaacaga tattaaatgt atcacatgaa gttacaaaca gtgattgtgt acttagcttt
     5281 ttcctctctg aggctgtgca acaaacagtt gaagaatcat cacctgtgta cctaggtgag
     5341 aagtttccag acaagaagcc taaggtgcag ttagtcatat cctacgagga tgtgaagctg
     5401 accatactag tgaaacacat gaaaaacatt catctcccag atggctctgc gcccagtgca
     5461 catgttgaat tttatctttt accatatccc agtgaagttc gtaggaggaa aacaaaatct
     5521 gttccaaaat gtacggaccc aacttacaat gaaattgtag tatatgatga agtcacagag
     5581 ctccaaggac atgtcttaat gcttattgtg aagagtaaaa ctgtatttgt gggagcaatt
     5641 aacatccgac tctgtagtgt cccactcgat aaagaaaaat ggtatccatt aggaaacagt
     5701 ataatttgac cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata
     5761 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     5821 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     5881 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5941 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     6001 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     6061 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     6121 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     6181 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     6241 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     6301 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     6361 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     6421 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6481 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6541 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     6601 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6661 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6721 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     6781 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6841 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     6901 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     6961 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     7021 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     7081 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     7141 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     7201 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     7261 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     7321 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     7381 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     7441 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     7501 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     7561 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     7621 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     7681 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     7741 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     7801 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     7861 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     7921 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     7981 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     8041 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     8101 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     8161 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     8221 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     8281 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     8341 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     8401 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     8461 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     8521 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     8581 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     8641 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     8701 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     8761 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     8821 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     8881 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8941 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     9001 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     9061 at
//
