LOCUS       sequence-391915-        6409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  visualization of clathrin-coated vesicle in living mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6409)
  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 6409)
  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: pHcRed-CLCa
FEATURES             Location/Qualifiers
     source          1..6409
                     /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             613..1296
                     /codon_start=1
                     /product="far-red fluorescent variant of Heteractis crispa 
                     chromoprotein"
                     /label=HcRed1
                     /note="mammalian codon-optimized"
                     /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH
                     VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT
                     AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL
                     KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD
                     LPEKAN"
     CDS             1309..1992
                     /codon_start=1
                     /product="far-red fluorescent variant of Heteractis crispa 
                     chromoprotein"
                     /label=HcRed1
                     /note="mammalian codon-optimized"
                     /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH
                     VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT
                     AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL
                     KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD
                     LPEKAN"
     polyA_signal    3198..3319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3235..3254)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3289..3308
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3326..3781)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3463..3484)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3675..3694
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3808..3912
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3914..4271
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3914..4105
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3937..3957)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4122..4257
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4306..5100
                     /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             4306..5100
                     /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(4360..4379)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4970..4989
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5288..5307)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5332..5379
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5707..6295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6196..6215
                     /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 cggcctgctg aaggagagta tgcgcatcaa gatgtacatg
      661 gagggcaccg tgaacggcca ctacttcaag tgcgagggcg agggcgacgg caaccccttc
      721 gccggcaccc agagcatgag aatccacgtg accgagggcg cccccctgcc cttcgccttc
      781 gacatcctgg ccccctgctg cgagtacggc agcaggacct tcgtgcacca caccgccgag
      841 atccccgact tcttcaagca gagcttcccc gagggcttca cctgggagag aaccaccacc
      901 tacgaggacg gcggcatcct gaccgcccac caggacacca gcctggaggg caactgcctg
      961 atctacaagg tgaaggtgca cggcaccaac ttccccgccg acggccccgt gatgaagaac
     1021 aagagcggcg gctgggagcc cagcaccgag gtggtgtacc ccgagaacgg cgtgctgtgc
     1081 ggccggaacg tgatggccct gaaggtgggc gaccggcacc tgatctgcca ccactacacc
     1141 agctaccgga gcaagaaggc cgtgcgcgcc ctgaccatgc ccggcttcca cttcaccgac
     1201 atccggctcc agatgctgcg gaagaagaag gacgagtact tcgagctgta cgaggccagc
     1261 gtggcccggt acagcgacct gcccgagaag gccaacagat ctcccgggat ggtgagcggc
     1321 ctgctgaagg agagtatgcg catcaagatg tacatggagg gcaccgtgaa cggccactac
     1381 ttcaagtgcg agggcgaggg cgacggcaac cccttcgccg gcacccagag catgagaatc
     1441 cacgtgaccg agggcgcccc cctgcccttc gccttcgaca tcctggcccc ctgctgcgag
     1501 tacggcagca ggaccttcgt gcaccacacc gccgagatcc ccgacttctt caagcagagc
     1561 ttccccgagg gcttcacctg ggagagaacc accacctacg aggacggcgg catcctgacc
     1621 gcccaccagg acaccagcct ggagggcaac tgcctgatct acaaggtgaa ggtgcacggc
     1681 accaacttcc ccgccgacgg ccccgtgatg aagaacaaga gcggcggctg ggagcccagc
     1741 accgaggtgg tgtaccccga gaacggcgtg ctgtgcggcc ggaacgtgat ggccctgaag
     1801 gtgggcgacc ggcacctgat ctgccaccac tacaccagct accggagcaa gaaggccgtg
     1861 cgcgccctga ccatgcccgg cttccacttc accgacatcc ggctccagat gctgcggaag
     1921 aagaaggacg agtacttcga gctgtacgag gccagcgtgg cccggtacag cgacctgccc
     1981 gagaaggcca acagaactcg agctcaagct agcttcgaat tcggcacgag cggtgggtcg
     2041 gttggttttt gtctcaccgt tggtgtccgt gccgttcagt tgcccgccat ggctgagctg
     2101 gatccgttcg gcgcccctgc cggcgcccct ggcggtcccg cgctggggaa cggagtggcc
     2161 ggcgccggcg aagaagaccc ggctgcggcc ttcttggcgc agcaagagag cgagattgcg
     2221 ggcatcgaga acgacgaggc cttcgccatc ctggacggcg gcgcccccgg gccccagccg
     2281 cacggcgagc cgccgggggg tccggatgct gttgatggag taatgaatgg tgaatactac
     2341 caggaaagta atggtccaac agacagttat gcagctattt cacaagtgga tcgattgcag
     2401 tcagagcctg aaagtatccg taaatggaga gaagaacaaa tggaacgctt ggaagccctt
     2461 gatgccaatt ctcggaagca agaagcagag tggaaagaaa aggcaataaa ggagctagaa
     2521 gaatggtatg caagacagga cgagcagcta cagaaaacaa aagcaaacaa cagggcagca
     2581 gaagaagcct ttgtaaatga cattgacgag tcgtccccag gcactgagtg ggaacgggtg
     2641 gcccggctgt gtgactttaa ccccaagtct agcaagcagg ccaaagatgt ctcccgcatg
     2701 cgctcagtcc tcatctccct caagcaggcc ccgctggtgc actgaagagc caccctgtgg
     2761 aaacactaca tctgcaatat cttaatccta ctcagtgaag ctcttcacag tcattggatt
     2821 aattatgttg agttcttttg gaccaaacct ttttgtcttt agagttgttc attgtttgtg
     2881 attgcatgtt tccttccttc aactgtgttc tccctggcat tcagagagga gggagaggag
     2941 gaagaggaag gggagggaag cttcccaaga gtagcctcaa cctgtgcttc tgtgcattat
     3001 tctgagaata aatttctgtt tcaaactgta aaaaaaaaaa aaaaaaactc gacggtaccg
     3061 cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     3121 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3181 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3241 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3301 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3361 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3421 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3481 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3541 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3601 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3661 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3721 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3781 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3841 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3901 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3961 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4021 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4081 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4141 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4201 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4261 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4321 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4381 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4441 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4501 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4561 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4621 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4681 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4741 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4801 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4861 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4921 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4981 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5041 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5101 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5161 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5221 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     5281 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5341 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5401 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5461 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagcaacgt
     5521 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     5581 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5641 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5701 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5761 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5821 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5881 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5941 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6001 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6061 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6121 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6181 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6241 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6301 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6361 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
