LOCUS       sequence-486255-        7278 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Encodes mouse VAV3 and N-terminal EGFP tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7278)
  AUTHORS   Qi Y, Yu CH
  TITLE     PI(3,4,5)P3-mediated Cdc42 activation regulates macrophage podosome 
            assembly.
  JOURNAL   Cell Mol Life Sci. 2025 Mar 24;82(1):127. doi: 
            10.1007/s00018-025-05664-2.
  PUBMED    40126693
REFERENCE   2  (bases 1 to 7278)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Mol 
            Life Sci."; date: "2025-03-24"; volume: "82(1):127. doi"; pages: " 
            10.1007/s00018-025-05664-2"
COMMENT     Sequence Label: EGFP-VAV3
FEATURES             Location/Qualifiers
     source          1..7278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     491..510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        765..1068
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1069..1272
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1223..1243
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1311..1320
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1317..1319,1320..1322,1323..2033)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1317..1319,1320..1322,1323..2033)
                     /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(1362..1383)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1623..1642)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1970..1991
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4581..4646
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    4770..4891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4807..4826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4861..4880
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4898..5353)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5035..5056)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5247..5266
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5380..5484
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5486..5843
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5486..5677
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5509..5529)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5694..5829
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5878..6672
                     /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             5878..6672
                     /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(5932..5951)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6542..6561
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6860..6879)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6904..6951
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
       61 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      121 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
      181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      661 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
      721 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      781 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      841 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      901 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      961 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     1021 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     1081 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     1141 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     1201 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     1261 ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg
     1321 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1381 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1441 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1501 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1561 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1621 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1681 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1741 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1801 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1861 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1921 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1981 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagatggagc
     2041 cgtggaagca gtgcgctcag tggctcatcc acagcaaggt gctgcccccc aaccatcggg
     2101 tgacctggga ctcggcgcag gtgttcgacc tggcgcagac ccttcgcgat ggagtcctgc
     2161 tctgccagct gctcaacaac ctgcggccac actccatcaa tctgaaggag atcaacctga
     2221 ggccccagat gtcccagttt ctctgtttga agaacatccg gacattcctg gccgcctgct
     2281 gcgatacctt tggaatgagg aaaagtgaac tttttgaggc atttgaccta tttgatgttc
     2341 gagactttgg aaaggttata gaaacattat ctcggctatc ccgaacacca atagcattgg
     2401 ccacaggaat caggccgttc cccactgaag aaagtataaa tgatgaagac atctacaaag
     2461 gccttcccga cttaatagat gaaacccgcg tggaagatga agaagacctc tatgactgtg
     2521 tgtacggcga ggatgaaggc ggagaagtct atgaagactt gatgaaggcg gaggaagccc
     2581 agcaacctaa atcacaagaa aatgatatac ggagttgttg cctagcagaa attaggcaga
     2641 cagaagaaaa atacacagaa acgttggagt caatagaaaa atatttcatg gcaccattga
     2701 aaagatttct gacggcggcg gaatttgatt cagtatttat taacattcct gaccttgtaa
     2761 aagttcatcg gagcttaatg caagagattc atgattccat tgtaaataaa gatgaccaga
     2821 acttgtatca agtttttatt aactacaagg aaagattggt tatttatggg cagtactgca
     2881 gtggagtgga gtcagccatc tctaacctag actacatctc taagacaaaa gaagatgtca
     2941 aactgaaatt agaggaatgt tcaaaaagag ccaataacgg gaagtttact ctgcgcgatt
     3001 tgctggtcgt tccaatgcag cgcgtgctga agtatcacct cctccttcag gagctggtaa
     3061 aacacaccca tgaccctatg gagaaggcga acctgaagct ggctctggat gcaatgaagg
     3121 acttggcaca atatgtgaat gaagtaaaaa gagataatga gacccttcgt gaaattaagc
     3181 agtttcagct atctatagag aatttgaacc agccagtttt gctctttgga cgaccacagg
     3241 gagacggtga aatccgaata accactctgg acaagcatac aaaacaagaa aggcatatct
     3301 tcttattcga cttggcagtg attgtctgta aaaggaaagg cgataactac gagatgaagg
     3361 aaataataga tctccagcag tacaagatag ccaacaaccc taccactgac aaagagaata
     3421 aaaagtggtc ttatggcttc tacctcatcc atacccaagg acaaaatggg ttagaatttt
     3481 attgcaaaac aaaagattta aagaaaaaat ggctggaaca gtttgaaatg gctctatcca
     3541 acataaggcc agactatgca gactccaatt tccatgattt caagatgcac accttcactc
     3601 gggtgacatc ctgcagggtc tgccagatgc tcctgagggg aacattttat caaggctact
     3661 tatgttttaa gtgtggagca aaagcacata aagaatgttt gggaagagta gacaattgtg
     3721 gcagagttaa ttccgttgaa caagggccat tcaaaccacc agagaaacgg accaatggac
     3781 ttcggagagc ttccagacag gtggacccag gtttgccgaa gatgcaggtc attaggaact
     3841 acactggcac cccggcccca gggcttcatg aaggaccacc attgcatatc caggcaggcg
     3901 acactgttga acttctgaga ggagatgcac acagtgtgtt ctggcaggga cgaaacttag
     3961 cttcaggaga ggttggattt ttcccaagtg atgctgtcaa accctctcca tgtgtaccta
     4021 aaccagtaga ttattcttgc caaccctggt atgctggccc aatggaaaga ttgcaggcag
     4081 aaactgaact tattaacagg gtgaatagta cttaccttgt aaggcacagg accaaagaat
     4141 ctggagaata tgcaattagc attaagtaca ataatgaagc aaaacacatc aagattttaa
     4201 caagagatgg cttttttcac attgcagaaa atagaaaatt taaaagctta atggaacttg
     4261 tggagtacta caagcaccat tctctcaaag aagggttcag gaccttagat acaacgctac
     4321 agtttccata caaggagcca gaacagccag ctggacagag gggtaataga acaggcaaca
     4381 gcttgttgag tcccaaagta ctgggtattg ccatcgctcg gtacgacttc tgtgcaagag
     4441 atatgcggga gctctcccta ttgaaaggag acatggtgaa gatttacacg aagatgagtg
     4501 caaatggctg gtggagaggt gaagtaaatg gcagggtggg ctggtttcca tccacatatg
     4561 tggaagaaga tgaaactagt tccggactca gatctcgagc tcaagcttcg aattctgcag
     4621 tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata
     4681 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     4741 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     4801 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4861 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     4921 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4981 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     5041 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5101 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5161 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5221 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5281 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5341 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5401 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5461 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     5521 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5581 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5641 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5701 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5761 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5821 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     5881 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5941 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     6001 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     6061 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     6121 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     6181 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     6241 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     6301 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     6361 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     6421 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     6481 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     6541 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     6601 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6661 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     6721 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     6781 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     6841 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     6901 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     6961 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     7021 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     7081 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     7141 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7201 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7261 aaaagatcaa aggatctt
//
