LOCUS       sequence-486254-        6600 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Encodes human ARHGEF6 and N-terminal EGFP tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6600)
  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 6600)
  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-ARHGEF6
FEATURES             Location/Qualifiers
     source          1..6600
                     /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"
     polyA_signal    4091..4212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4128..4147)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4182..4201
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4219..4674)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4356..4377)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4568..4587
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4701..4805
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4807..5164
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4807..4998
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4830..4850)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5015..5150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5199..5993
                     /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             5199..5993
                     /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(5253..5272)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5863..5882
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6181..6200)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6225..6272
                     /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 agtccggact
     2041 cagatctcga gctcaagctt ccatgacgga aaatggaagt catcagttga tagtaaaagc
     2101 aagattcaac tttaagcaga ctaatgagga tgaactgtca gtttgtaagg gggacatcat
     2161 ttacgtcaca cgagttgaag aaggaggctg gtgggaaggc acattaaatg ggagaacagg
     2221 ctggttcccc agtaattatg tccgtgaaat taaatccagt gagagacctc tctccccaaa
     2281 agccgtcaaa ggatttgaaa ctgctccact taccaagaat tattatactg tggtgttaca
     2341 gaacatcctg gacactgaaa aagaatatgc taaagaactt cagtctcttc ttgttactta
     2401 cttaagaccc ctgcagtcca ataacaatct gagtactgtg gaggttacat ctttactggg
     2461 aaacttcgag gaagtatgca catttcaaca gacactctgc caagccttgg aagaatgttc
     2521 aaagtttcca gaaaaccagc acaaagtagg aggttgtcta ctgagtctca tgcctcattt
     2581 taaatctatg tatctggctt actgtgcaaa ccatccttca gctgtaaatg tgctcactca
     2641 gcacagtgat gagttggaac aattcatgga aaatcaaggt gcatcgagcc caggtatcct
     2701 cattttaaca acaaacctca gcaaaccatt catgcgactg gagaaatatg ttactctctt
     2761 gcaagagtta gaacggcata tggaggatac tcatccagat catcaggata ttctgaaagc
     2821 aatcgtagca ttcaaaactc tcatggggca atgtcaagat ctgaggaaga gaaaacagct
     2881 ggagttacag atactgtccg aacctattca ggcatgggaa ggagaagata ttaaaaactt
     2941 gggaaatgtg atttttatgt cacaagtaat ggtgcagtat ggagcatgtg aggaaaaaga
     3001 ggagcggtac cttatgttat tttcaaatgt cctgataatg ttatctgcaa gtcctcggat
     3061 gagtggcttt atctatcagg gaaaaatacc aatagcagga acggtggtga ctagattaga
     3121 tgaaattgaa gggaatgact gcacatttga aatcactggt aacacagtgg agagaattgt
     3181 ggtccattgt aacaacaacc aggacttcca ggaatggttg gagcagctga acagactgat
     3241 cagaggacct gcctcttgca gttcattatc caaaacctca tcgtcatcat gtagtgctca
     3301 ttcttctttt agctctaccg gacagccccg aggacccttg gagcctcctc aaattataaa
     3361 accgtggagt ttaagttgtc tacgacctgc acctccactt agaccatcag cagcactagg
     3421 ttataaagag aggatgtctt atatcttaaa ggagtctagt aaaagcccta aaacgatgaa
     3481 gaaatttctt cataaaagga agactgagag aaaaccatcg gaggaggaat atgtgattag
     3541 gaaaagtaca gctgctctgg aagaggatgc tcaaatcctt aaagtgatcg aagcctactg
     3601 caccagcgca aattttcaac aaggccatgg ctcaagtact cgaaaagatt ccattccaca
     3661 agtcctactc cctgaggaag agaaactcat cattgaagaa accagaagca acggccagac
     3721 catcatggaa gaaaagagcc ttgttgatac tgtttacgcc ttgaaggacg aggtcagaga
     3781 actgaagcag gaaaataaaa gaatgaagca atgcctggaa gaagaactga aatcaagaag
     3841 ggacctagaa aagctggtgc ggaggctttt gaagcaaaca gatgagtgta ttcgaggcga
     3901 gtccagtagc aagacctcaa ttcttccata gaattctgca gtcgacggta ccgcgggccc
     3961 gggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     4021 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     4081 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4141 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4201 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4261 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4321 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4381 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4441 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4501 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4561 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4621 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4681 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4741 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4801 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4861 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4921 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4981 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5041 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5101 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5161 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5221 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5281 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5341 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     5401 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5461 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5521 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5581 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5641 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5701 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     5761 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5821 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5881 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5941 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6001 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6061 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6121 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     6181 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     6241 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     6301 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     6361 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     6421 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6481 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6541 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
