LOCUS       sequence-486261-        7086 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Encodes human FGD3 and N-terminal EGFP tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7086)
  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 7086)
  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-FGD3
FEATURES             Location/Qualifiers
     source          1..7086
                     /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"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1143..1277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1180..1199)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1234..1253
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1316..1340
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3528..3552)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3574..4284,4285..4287,4288..4290))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3574..4284,4285..4287,4288..4290))
                     /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(3616..3637)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3965..3984
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4224..4245
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4287..4296
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4362..4380)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4363..4380)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(4415..5399)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      5470..5925
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5557..5576)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5767..5788
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5951..6055
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6056..6124,6125..6916)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6274..6293)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
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 taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgctccc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tgggtaccgg gcccaatgca ttggcgccgc ggccgcgaat
     1081 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1141 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1201 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1261 cttatcatgt ctggatctac gtaatacgac tcactatagt tctagaggct cgatcaccac
     1321 tttgtacaag aaagctgggt ggattcacgg agcagctgcg cccataggga cctgaagctg
     1381 cagggccccc gggctgtcct gggccgtgtc cccatgggca gcagtgctta gggtttccag
     1441 ccactgctgc tgcagctctg cggaggaggc gctcaggtac caggactgct tggcccactg
     1501 cagcttccac acatgccccg agtccagcct ctcctcaggg tccggcacac tcagtttgca
     1561 gctggggaga gggatggtgc ggggcagccg gccgtcctgg ctgcctccct gcaggtgcag
     1621 cacctgggga tctgacatgg ggatggcggc ccacacctcg ctccaggtct caccgctctc
     1681 tgacagccgc agggggccgc agagcaggct gggctggggg tctgcagtgg gtgtcttctc
     1741 tgtgctctca ggggccactg gctgtgtcag gaaacaatct ctgcagacac ggctctgccg
     1801 gctgttctcg gccttgaact cggagcactt cccacagatg accgccccac acagcttgca
     1861 gtgatgcctc ctcttggtga tggagttgaa ggtctcacca cagctcttac agctctgctt
     1921 ctccttgtca cgtctggtct tagaggatag ttttctgggc tcgagccctg ctgcacccga
     1981 actgccttcg gtggtcacca caggctccac agggctggtg ctcgtgatag gcatgtctgg
     2041 agagaggctg gggtcctcat cctggctaaa ggcgccacca aaagccttga aggtttcgct
     2101 gttctgtttg tgcttctcga tggtggcctg gatgatctga atccattctt tcttctcttc
     2161 ctctgtccgc gtctgcagct ccagggacct ttttcttccc gttatgatga atgtatgtgc
     2221 tgtgtttggc ttgacgatat cctgcacctg gaggcctgag atgtccatct tctcccggac
     2281 gctgaacttc tggcccatga gccgcagctt gggcacacag taaaggatca tgctgttgaa
     2341 caggaagagg tggcggtcct ggggggtgcc gttcttggct gacagtttct ggatttggcc
     2401 ctccttgatc agttcattgg ccgggttgac aatgtcttct tccccaccca gctgctcgta
     2461 cacctccaag agcttgtgca ttttctccac tttccgaatg gcagcattgg agtggttggc
     2521 ggctgtggag atgagctcca aggacctctc cgcatccttc cggtctgggg cgtcctgcgg
     2581 gagcctcttc agatagtcct tgagcagcag ctcgtaccgg gggaccctct gcacgggctc
     2641 cagcatgtgg tgctgcagcg tcaggttccc gcatacctcc tgcttctgga tgctgtggat
     2701 gacgtcttta aacagtgggg agcgctgggt ccacgtgctc accagcccta cggctcggtc
     2761 aaagttcttg acatactcgc cgtacatctt caggaatggg gccagcttct gcaggatgtc
     2821 cccgagccgt gggtttgtgt cccactcctc cgtgatccgc gtcttcagct ccggcagcag
     2881 gaactgcccg tggaagcggt ggatggagga gatgttagag aatatgccca tgatgacttc
     2941 tggagggatc cccgcatccg tcagcctggt gcagaaaacc tggtccagca ggtgcagccg
     3001 cttcacatag gtctcctcgg tgtgcaggag ctcctgggca atgtggagaa gcttctgggg
     3061 gccagagtgc tgggccaggc cggcatcctt gtcagccttc tggggggtgt tctcagagtc
     3121 aggttcctca cccacgtcgc tgtccgcctc ctcctggggg gtgaccttcg ggacctggct
     3181 gtccagggcc atctctgcgt gcgcccccag tacagtgggg cttgggccag cctccaagcc
     3241 ctcctcgcag ggaaagttct ctccagccac actggaggat ggactgtcga tcccgctgtc
     3301 ccggttgggg atctttaagc taccactcag ctctcccgtg gggcctatgt ctggagagcc
     3361 gtcccctgct gcagccaggc tgacagggtc cccacagtgg gctctgggcc caacagggag
     3421 cgcctgaagc ttccctaggg aggaactgcc aggcccagtg tctggcatcc ctagggcagc
     3481 aatgggtcct ggaggggttg aggaccccct gcctgactcc atggtggagc ctgctttttt
     3541 gtacaaactt gtgattcgag atctgagtcc ggacttgtac agctcgtcca tgccgagagt
     3601 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     3661 tttgctcagg gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc
     3721 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt
     3781 gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac
     3841 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     3901 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     3961 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     4021 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca
     4081 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     4141 ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     4201 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     4261 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta gccttgaatt
     4321 cgaatcgatg ggatcctgca aaaagaacaa gtagcttgta ttctatagtg tcacctaaat
     4381 caagcttgct ccgacgtccc caggcagaat ggcggttccc taaacgagca ttgcttatat
     4441 agacctccca ttaggcacgc ctaccgccca tttacgtcaa tggaacgccc atttgcgtca
     4501 ttgcccctcc ccattgacgt caatggggat gtacttggca gccatcgcgg gccatttacc
     4561 gccattgacg tcaatgggag tactgccaat gtaccctggc gtacttccaa tagtaatgta
     4621 cttgccaagt tactattaat agatattgat gtactgccaa gtgggccatt taccgtcatt
     4681 gacgtcaata gggggcgtga gaacggatat gaatgggcaa tgagccatcc cattgacgtc
     4741 aatggtgggt ggtcctattg acgtcaatgg gcattgagcc aggcgggcca tttaccgtaa
     4801 ttgacgtcaa tgggggaggc gccatatacg tcaataggac cgcccatatg acgtcaatag
     4861 gaaagaccat atatagagac cattgacgtc aatgggggag tggctatggg cggtattagg
     4921 aagccccata tatggtatat gggaccgccc attgggaggg gctatctacg tcaataggaa
     4981 aacccatata tggaatacta tatggcatag ggccaataca tagtattgaa cctggccaat
     5041 agccatattg gcatagggcc atattggata ttgcctatat attgatcctg gcatatagcc
     5101 aatatggccg ccattattgg caccatgcca attcaatatg gcggacttgg cacagggcca
     5161 agacccctcc tcaaacttgg caccgtgcca agtagacccc tccccagttg gcacagtgcc
     5221 aggtccgcca tattgaattg gcatggtgcc aagtccgcca tattgaattg gctggggtcc
     5281 agatccacca tattgaattg gcacgagtcc atatacgcca tattgaattg gcacaggtcc
     5341 agatccacca tattgaattg gcatgagtcc atatacgcca tattgaattg gcggtcgact
     5401 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     5461 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5521 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5581 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5641 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5701 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5761 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5821 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5881 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     5941 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6001 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6061 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6121 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6181 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6241 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6301 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6361 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6421 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6481 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6541 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6601 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6661 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6721 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6781 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6841 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6901 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6961 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7021 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7081 atcttc
//
