LOCUS       sequence-484516-        7272 bp    DNA     circular SYN 27-AUG-2025
DEFINITION  Encodes human VAV1 and N-terminal EGFP tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7272)
  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 7272)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 27, 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-VAV1
FEATURES             Location/Qualifiers
     source          1..7272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1310..1319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1316..1318,1319..1321,1322..2032)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1316..1318,1319..1321,1322..2032)
                     /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(1361..1382)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1622..1641)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1969..1990
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4574..4639
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    4763..4884
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4800..4819)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4854..4873
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4891..5346)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5028..5049)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5240..5259
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5373..5477
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5479..5836
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5479..5670
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5502..5522)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5687..5822
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5871..6665
                     /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             5871..6665
                     /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(5925..5944)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6535..6554
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6853..6872)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6897..6944
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
     1321 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1381 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1441 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1501 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1561 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1621 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1681 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1741 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1801 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1861 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1921 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1981 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agatggagct
     2041 gtggcgccaa tgcacccact ggctcatcca gtgccgggtg ctgccgccca gccaccgcgt
     2101 gacctgggat ggggctcagg tgtgtgaact ggcccaggcc ctccgggatg gtgtccttct
     2161 gtgtcagctg cttaacaacc tgctacccca tgccatcaac ctgcgtgagg tcaacctgcg
     2221 cccccagatg tcccagttcc tgtgccttaa gaacattaga accttcctgt ccacctgctg
     2281 tgagaagttc ggcctcaagc ggagcgagct cttcgaagcc tttgacctct tcgatgtgca
     2341 ggattttggc aaggtcatct acaccctgtc tgctctgtcc tggaccccga tcgcccagaa
     2401 cagggggatc atgcccttcc ccaccgagga ggagagtgta ggtgatgaag acatctacag
     2461 tggcctgtcc gaccagatcg acgacacggt ggaggaggat gaggacctgt atgactgcgt
     2521 ggagaatgag gaggcggaag gcgacgagat ctatgaggac ctcatgcgct cggagcccgt
     2581 gtccatgccg cccaagatga cagagtatga caagcgctgc tgctgcctgc gggagatcca
     2641 gcagacggag gagaagtaca ctgacacgct gggctccatc cagcagcatt tcttgaagcc
     2701 cctgcaacgg ttcctgaaac ctcaagacat tgagatcatc tttatcaaca ttgaggacct
     2761 gcttcgtgtt catactcact tcctaaagga gatgaaggaa gccctgggca cccctggcgc
     2821 agccaatctc taccaggtct tcatcaaata caaggagagg ttcctcgtct atggccgcta
     2881 ctgcagccag gtggagtcag ccagcaaaca cctggaccgt gtggccgcag cccgggagga
     2941 cgtgcagatg aagctggagg aatgttctca gagagccaac aacgggaggt tcaccctgcg
     3001 ggacctgctg atggtgccta tgcagcgagt tctcaaatat cacctccttc tccaggagct
     3061 ggtgaaacac acgcaggagg cgatggagaa ggagaacctg cggctggccc tggatgccat
     3121 gagggacctg gctcagtgcg tgaacgaggt caagcgagac aacgagacac tgcgacagat
     3181 caccaatttc cagctgtcca ttgagaacct ggaccagtct ctggctcact atggccggcc
     3241 caagatcgac ggggaactca agatcacctc ggtggaacgg cgctccaaga tggacaggta
     3301 tgccttcctg ctcgacaaag ctctactcat ctgtaagcgc aggggagact cctatgacct
     3361 caaggacttt gtaaacctgc acagcttcca ggttcgggat gactcttcag gagaccgaga
     3421 caacaagaag tggagccaca tgttcctcct gatcgaggac caaggtgccc agggctatga
     3481 gctgttcttc aagacaagag aattgaagaa gaagtggatg gagcagtttg agatggccat
     3541 ctccaacatc tatccggaga atgccaccgc caacgggcat gacttccaga tgttctcctt
     3601 tgaggagacc acatcctgca aggcctgtca gatgctgctt agaggtacct tctatcaggg
     3661 ctaccgctgc catcggtgcc gggcatctgc acacaaggag tgtctgggga gggtccctcc
     3721 atgtggccga catgggcaag atttcccagg aactatgaag aaggacaaac tacatcgcag
     3781 ggctcaggac aaaaagagga atgagctggg tctgcccaag atggaggtgt ttcaggaata
     3841 ctacgggctt cctccacccc ctggagccat tggacccttt ctacggctca accctggaga
     3901 cattgtggag ctcacgaagg ctgaggctga acagaactgg tgggagggca gaaatacatc
     3961 tactaatgaa attggctggt ttccttgtaa cagggtgaag ccctatgtcc atggccctcc
     4021 tcaggacctg tctgttcatc tctggtacgc aggccccatg gagcgggcag gggcagagag
     4081 catcctggcc aaccgctcgg acgggacttt cttggtgcgg cagagggtga aggatgcagc
     4141 agaatttgcc atcagcatta aatataacgt cgaggtcaag cacattaaaa tcatgacagc
     4201 agaaggactg taccggatca cagagaaaaa ggctttccgg gggcttacgg agctggtgga
     4261 gttttaccag cagaactctc taaaggattg cttcaagtct ctggacacca ccttgcagtt
     4321 ccccttcaag gagcctgaaa agagaaccat cagcaggcca gcagtgggaa gcacaaagta
     4381 ttttggcaca gccaaagccc gctatgactt ctgcgcccga gaccgatcag agctgtcgct
     4441 caaggagggt gacatcatca agatccttaa caagaaggga cagcaaggct ggtggcgagg
     4501 ggagatctat ggccgggttg gctggttccc tgccaactac gtggaggaag attattctga
     4561 atactgcact agttccggac tcagatctcg agctcaagct tcgaattctg cagtcgacgg
     4621 taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt
     4681 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     4741 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     4801 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4861 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
     4921 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4981 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     5041 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     5101 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     5161 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     5221 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     5281 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     5341 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5401 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5461 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     5521 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5581 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5641 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     5701 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     5761 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5821 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     5881 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5941 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     6001 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     6061 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     6121 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     6181 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     6241 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     6301 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     6361 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     6421 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     6481 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     6541 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     6601 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     6661 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     6721 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6781 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     6841 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     6901 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     6961 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     7021 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     7081 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     7141 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     7201 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     7261 caaaggatct tc
//
