LOCUS       sequence-486259-        6873 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Encodes human FGD2 and N-terminal EGFP tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6873)
  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 6873)
  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-FGD2
FEATURES             Location/Qualifiers
     source          1..6873
                     /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"
     primer_bind     744..761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    878..899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(914..919,920..937,938..944)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    952..968
                     /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     957..979
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     976..992
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     976..992
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1011..1031
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1013..1031
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1144..1278
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1181..1200)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1235..1254
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1317..1341
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3316..3340)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3362..4072,4073..4075,4076..4078))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3362..4072,4073..4075,4076..4078))
                     /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(3404..3425)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3753..3772
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4012..4033
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4075..4084
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4150..4168)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4151..4168)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(4203..5187)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      5258..5713
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5345..5364)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5555..5576
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5739..5843
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5844..5912,5913..6704)
                     /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(6062..6081)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
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 ttaccgcctt tgagtgagct gataccgctc
      721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
      781 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
      841 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
      901 aggcacccca ggctttacac tttatgctcc cggctcgtat gttgtgtgga attgtgagcg
      961 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
     1021 ctcactaaag ggaacaaaag ctgggtaccg ggcccaatgc attggcgccg cggccgcgaa
     1081 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     1141 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1201 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1261 tcttatcatg tctggatcta cgtaatacga ctcactatag ttctagaggc tcgatcacca
     1321 ctttgtacaa gaaagctggg tccaagtcgg acaggtcccc atcgttgggc cagctggggc
     1381 tccagccact ggccgcccgc tccatggcct tcacccagcg gcccttcagc tcctccgtct
     1441 cggccttgaa ggtgtagagc tggcctgact gctgtagctg gaagacccga gggtccccct
     1501 ggggcccaac agtcacctgg tagcccagca gggggatgga ggtgtgagcc ctcatgtcct
     1561 gaggggcagc atagacatag agcacgaggg ggtcatcccg agggatcaca caccagcccc
     1621 gggggccgct cttgccccac ttgtccccga tgagctgcag gaagctgcac atcaggctct
     1681 ggtcaggcgt ggctgaggac cctttctcca ggatgccccg cctcttgtcc tccttggcct
     1741 caggcagcac atttccagtg aggaatgcgt agcagtggag gcagactcgg ttgggcctgt
     1801 tgtcgtcgta tttcagttcg gcccggtagt cggagcacct ggcacacacc acatagccgc
     1861 aggcccggca gtggtggcga cggcgcgtca gagcgttgaa gggctcctgg cagcgcatgc
     1921 acatggtcac catcttgtcc cggacccact gcggtgcccg gaggcccagc tcctcagact
     1981 gcagctcctg ctcctggatg tctccctcag gcccctgggc cgcagccttg aaggtttcat
     2041 tccgcttctc gatttggtca atggctgctt ggaaggcctg catccaggaa atcatttcct
     2101 cctgggaccg ggcttgcagc tccagggtgc gctgcttccc ggacaccagg aaggagtggg
     2161 gaaactcagc atccatcagc tcccgcacct tcatcccggc cacatcgatg cgggtcctca
     2221 cctggaactg ggcgcccacc tggatcaccc tgggcacaca gtagagcagc atgttgttga
     2281 acaagaaaag gtagcgctcc atggggtcgt tgcggcggaa ggagatcttg aggaccgggc
     2341 cctcacggag cagggtgtta gaggggtcta ctatgtcgtc ctcgaggccc aggcgctggt
     2401 acacctccca caggtcctgc agccgctcca tctcagtgat ggctgcattg gagtgctggg
     2461 cagctgagaa gatcatgtcc agggctttct gggcatcggc ctggtctggg gcctgggctg
     2521 gcagcttctg gatgtactcc ttgagcagca gctcgtaacg tggaattctc tgcactggtt
     2581 ccagcatgtg gtgctgcagg gtcaggctgc ccgaagcctc gctgctctgg atgcgagtga
     2641 gaacctcctg gaagagtgga gacttgtcgg tccaggtggc cagcagctca gccgctcgct
     2701 caaagttctt gacatactca ctgtacatct tcaggaaggg ggccagcttc tggatcacgt
     2761 caccgatgcg ggggttagct gtccagtcgt ccaggcgccg ctgcagctct gggaggaaga
     2821 actgagaatg gaactgatag atggaggaga tgttggagaa gatgaccctg accacatcct
     2881 ctgggaaggc cttgctgctg cgggctgtct tcagcagctc ctggaaaaac acctggtcta
     2941 gcaggtggag gcgcgccaca taggcctgct ctgtctccag cagctcctgg acgatcttct
     3001 tctctggctc ctgcgtcccc gatcctgaga gggtcacagg ctggcaagat ttcctccagg
     3061 cttcgctgga cagcttgttc ttcagggagt tcaggtacct cctgctgact gtcctgggct
     3121 cagaccccac ggcctccccg acattcgtct tctcccgtgg tcctggggac tcgggaggcc
     3181 tgcactcagg gcggtgatgc acgtcctcta gcctctggcc tctgggtgct gcttctgggg
     3241 tcctgctatt ctcaaacaca gtgaccaggt tggacacaga tgccagcttc tcctcacttg
     3301 cccccttcat ggtggagcct gcttttttgt acaaacttgt gattcgagat ctgagtccgg
     3361 acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     3421 ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt
     3481 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     3541 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
     3601 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     3661 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     3721 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     3781 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     3841 tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg
     3901 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     3961 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     4021 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     4081 tggcgaccgg tagcgctagc cttgaattcg aatcgatggg atcctgcaaa aagaacaagt
     4141 agcttgtatt ctatagtgtc acctaaatca agcttgctcc gacgtcccca ggcagaatgg
     4201 cggttcccta aacgagcatt gcttatatag acctcccatt aggcacgcct accgcccatt
     4261 tacgtcaatg gaacgcccat ttgcgtcatt gcccctcccc attgacgtca atggggatgt
     4321 acttggcagc catcgcgggc catttaccgc cattgacgtc aatgggagta ctgccaatgt
     4381 accctggcgt acttccaata gtaatgtact tgccaagtta ctattaatag atattgatgt
     4441 actgccaagt gggccattta ccgtcattga cgtcaatagg gggcgtgaga acggatatga
     4501 atgggcaatg agccatccca ttgacgtcaa tggtgggtgg tcctattgac gtcaatgggc
     4561 attgagccag gcgggccatt taccgtaatt gacgtcaatg ggggaggcgc catatacgtc
     4621 aataggaccg cccatatgac gtcaatagga aagaccatat atagagacca ttgacgtcaa
     4681 tgggggagtg gctatgggcg gtattaggaa gccccatata tggtatatgg gaccgcccat
     4741 tgggaggggc tatctacgtc aataggaaaa cccatatatg gaatactata tggcataggg
     4801 ccaatacata gtattgaacc tggccaatag ccatattggc atagggccat attggatatt
     4861 gcctatatat tgatcctggc atatagccaa tatggccgcc attattggca ccatgccaat
     4921 tcaatatggc ggacttggca cagggccaag acccctcctc aaacttggca ccgtgccaag
     4981 tagacccctc cccagttggc acagtgccag gtccgccata ttgaattggc atggtgccaa
     5041 gtccgccata ttgaattggc tggggtccag atccaccata ttgaattggc acgagtccat
     5101 atacgccata ttgaattggc acaggtccag atccaccata ttgaattggc atgagtccat
     5161 atacgccata ttgaattggc ggtcgactgg cgtaatagcg aagaggcccg caccgatcgc
     5221 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     5281 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5341 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5401 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5461 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5521 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5581 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5641 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5701 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5761 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5821 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5881 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5941 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6001 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6061 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6121 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6181 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6241 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6301 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6361 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6421 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6481 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6541 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6601 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6661 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6721 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6781 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6841 cgtcagaccc cgtagaaaag atcaaaggat ctt
//
