LOCUS       sequence-391928-        7751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  visualization of clathrin-coaed vesicle formation in living 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7751)
  AUTHORS   Sakaushi S, Inoue K, Zushi H, Senda-Murata K, Fukada T, Oka S, 
            Sugimoto K
  TITLE     Dynamic behavior of FCHO1 revealed by live-cell imaging microscopy: 
            its possible involvement in clathrin-coated vesicle formation.
  JOURNAL   Biosci Biotechnol Biochem. 2007 Jul;71(7):1764-8. doi: 
            10.1271/bbb.60720. Epub 2007 Jul 7.
  PUBMED    17617719
REFERENCE   2  (bases 1 to 7751)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1271/bbb.60720";
            journalName: "Biosci Biotechnol Biochem"; date: "2007-07"; volume: 
            "71"; issue: "7"; pages: "1764-8"
COMMENT     Sequence Label: pEGFP-FCHO1
FEATURES             Location/Qualifiers
     source          1..7751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4539..4660
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4576..4595)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4630..4649
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4667..5122)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4804..4825)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5016..5035
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5149..5253
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5255..5612
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5255..5446
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5278..5298)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5463..5598
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5647..6441
                     /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             5647..6441
                     /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(5701..5720)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6311..6330
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6629..6648)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6673..6720
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7049..7637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7538..7557
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgactt cgaattcccg ggtcgaccca cgcgtccgat
     1381 cttgacaggc actggacggg gcctgcaggg gtctccacag agaccatcag gatgtcgtat
     1441 tttggggagc atttttgggg cgagaaaaat catggctttg aggtcctgta ccacagcgtg
     1501 aagcaggggc ccatctccac caaggagctg gcggacttca tccgggagag ggccaccatc
     1561 gaggagacct actcgaaggc gatggcgaaa ctctccaagc tggccagcaa cgggaccccc
     1621 atggggacct tcgccccgct ctgggaggtc ttccgcgtct cctcggacaa gctggcgctg
     1681 tgccacctgg aactgacacg gaagttacag gatctcatca aggacgttct ccgctacggc
     1741 gaggaacagc tcaagaccca caagaagtgc aaggaggaag tggtgagcac cttggatgct
     1801 gtgcaggtac tctcgggcgt cagccagctc ctgcccaagt cccgcgagaa ctacctgaac
     1861 cgttgcatgg accaggagcg gctgcggagg gagagtacca gccagaagga gatggacaag
     1921 gcggagacta aaaccaagaa ggcggcagag agcctgcggc gctcagtgga aaaatacaac
     1981 tcagcccgag ctgactttga gcagaagatg ctggactcag ccctgcgctt ccaagccatg
     2041 gaggagacac acctgaggca catgaaggca ctgctgggct catatgctca ctcggtggag
     2101 gacacgcacg tgcagattgg gcaggtgcat gaggaattta agcagaacat cgagaacgtc
     2161 agcgtggaga tgctactcag gaagtttgca gagagtaagg gcacaggccg ggagaagcct
     2221 ggacctctgg acttcgaggc atacagtgcg gctgccctgc aggaagcgat gaaacgtttg
     2281 cggggagcca aggcctttcg ccttccagga ctaagccggc gggagcggga gccagagcca
     2341 cctgcagctg tagatttcct ggagcctgat tcagggacat gtccagaggt ggatgaagaa
     2401 ggtttcactg tccggcctga tgtgacccag aacagcacgg ccgagccctc ccgtttctcg
     2461 tccagcgact ccgacttcga cgatgaagag ccccgcaagt tctatgtgca catcaagcct
     2521 gccccggccc gggctccagc ctgcagcccc gaggcagcag cggcacagct cagggccacc
     2581 gcgggcagcc tcatccttcc tcctggccca gggggcacca tgaaacgcca ttcttcacgg
     2641 gacgctgctg ggaaacccca gagacctcgg tctgccccca gaaccagcag ctgtgcagag
     2701 agattgcagt cagaggagca ggtgtccaag aacctctttg ggccgcccct ggagtcagcc
     2761 tttgaccacg aagattttac aggctctagc agcctgggct tcacctccag cccctcccct
     2821 ttctcctcct cgtcgcccga aaacgtggag gattccggcc tggactctcc gtcccacgcg
     2881 gcacctggcc cctccccaga ttcctgggtc ccccgcccag gcaccccgca gagcccgccc
     2941 agctgtaggg cgccaccccc agaggccagg ggtatccggg caccgcctct gccagactcg
     3001 ccgcagcccc tcgcctcgtc tccaggcccc tgggggctgg aggccttggc cggaggagac
     3061 ctgatgcctg cacctgctga ccccacagcc agggagggcc tggcagcccc acccaggaga
     3121 cttcgctcta ggaaggtgtc ctgccctctc acacgtagca atggggacct gtctcgttcc
     3181 ctgagcccct ccccactggg ctcttcagcc gccagcactg ccttggaacg gcccagcttc
     3241 ttatcccaga caggacacgg agtctcccgg ggtccgagcc ctgtggtcct gggctcccag
     3301 gatgccctgc ccatagccac agccttcacg gagtatgtcc acgcctactt ccgtggccac
     3361 agccccagct gcctggctcg agtaactggg gagctgacca tgaccttccc tgctggcatc
     3421 gtgcgtgtgt tcagcgggac cccaccacca cctgtcctca gcttccggct tgtacacaca
     3481 accgctattg agcacttcca gcccaacgcc gatctgctgt tcagtgaccc ctcccagagt
     3541 gaccctgaga ccaaagactt ctggctcaac atggcagctc tgaccgaagc cctgcagcgc
     3601 caggcagagc agaaccccac tgcctcctac tacaacgtgg tgctgctgcg ataccagttc
     3661 tcccgcccgg gtccccagtc tgtgcctctg cagctcagtg cccactggca gtgtggagcc
     3721 accctcaccc aggtctcagt ggagtacggc taccggcccg gtgccacggc tgtgcccaca
     3781 ccactcacga acgtccagat cctgctgcct gtgggggagc ctgtgaccaa cgtccgcttg
     3841 cagccggctg ccacctggaa cctggaggag aagcggctca cttggaggct tccagatgtg
     3901 tccgaggcag gcggttctgg ccgcctctct gccagctggg agccgctctc agggcccagc
     3961 acacccagcc ccgtggctgc acagttcacc agcgagggga ccactctgtc gggcgtggac
     4021 ttggaactgg tgggcagcgg ttaccgcatg tcgctggtga agaggaggtt tgccacaggg
     4081 atgtacctgg tgagctgctg aacccgcaaa tgctgctgcc ccagctctac actgcgccct
     4141 ggtgctggct gaccaccccc tgccctcctg ccggaccctg gggcctccca ccccagcctc
     4201 cctgaggccc atactccacg gagaggagcc ccatgcccag cctggctgag cccgagattc
     4261 gctcctcccc ctcatgccaa ccccacacag gtcccggcct tttaatgttc tttgaataaa
     4321 cactttattt tctaataaaa taaaaaagga aacctttaaa aaaaaaaaaa aaaaaaaaaa
     4381 aaaaaaaggg cggccgctaa ttagttaggg gatccaccgg atctagataa ctgatcataa
     4441 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     4501 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     4561 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4621 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     4681 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4741 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4801 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4861 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4921 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4981 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     5041 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     5101 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5161 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5221 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     5281 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5341 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5401 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5461 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5521 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     5581 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     5641 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5701 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5761 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5821 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5881 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5941 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6001 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6061 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     6121 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     6181 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6241 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6301 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6361 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6421 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     6481 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6541 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6601 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     6661 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     6721 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     6781 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     6841 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     6901 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6961 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7021 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7081 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7141 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7201 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7261 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7321 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7381 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7441 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7501 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7561 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7621 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7681 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7741 ccgccatgca t
//
