LOCUS       sequence-410643-        7785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of hEFA6A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7785)
  AUTHORS   Ueda T, Hanai A, Takei T, Kubo K, Ohgi M, Sakagami H, Takahashi S, 
            Shin HW, Nakayama K
  TITLE     EFA6 activates Arf6 and participates in its targeting to the 
            Flemming body during cytokinesis.
  JOURNAL   FEBS Lett. 2013 Jun 5;587(11):1617-23. doi: 
            10.1016/j.febslet.2013.03.042. Epub 2013 Apr 18.
  PUBMED    23603394
REFERENCE   2  (bases 1 to 7785)
  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: 
            "10.1016/j.febslet.2013.03.042"; journalName: "FEBS Lett"; date: 
            "2013-06-5- 5"; volume: "587"; issue: "11"; pages: "1617-23"
COMMENT     Sequence Label: pEGFP-C1-hEFA6A
FEATURES             Location/Qualifiers
     source          1..7785
                     /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    4573..4694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4664..4683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4701..5156)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4838..4859)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5050..5069
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5183..5287
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5289..5646
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5289..5480
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5312..5332)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5497..5632
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5681..6475
                     /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             5681..6475
                     /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(5735..5754)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6345..6364
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6663..6682)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6707..6754
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7083..7671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7572..7591
                     /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 atctaccatg gcccagggtg ccatgcgctt ctgctcggaa
     1381 ggcgactgtg ccatctcccc accacgatgc ccacgccgct ggctccccga aggcccggtg
     1441 ccccagagcc ccccagccag catgtatggc agcacaggct ccttgctacg gcgagtggca
     1501 ggtccaggtc ctcgaggccg ggaactggga cgtgtgacag caccctgtac acctctgcgt
     1561 ggccccccct caccccgtgt tgctccctca ccctgggcac cctcttcacc cactgggcag
     1621 cccccacccg gggcccagag ctctgtggtc atcttccgct ttgtggagaa ggccagtgtg
     1681 aggccactga atgggctacc tgctccaggg ggcttgagtc ggagctggga tctgggtggg
     1741 gtttctcctc ccaggcccac cccagccctt gggcctggct ccaaccggaa gttacggctg
     1801 gaagcatcca catcagaccc actccccgcc agaggaggct cggccctacc tggcagccgg
     1861 aaccttgtac atgggccgcc agccccaccc caggttggag cagatggcct ttactcctct
     1921 ctccccaatg ggctgggggg cccccctgag cgcctggcca cactcttcgg aggacctgct
     1981 gacactggat tcctgaacca gggggatacc tggtcctccc cccgggaagt ctcctctcat
     2041 gcccagagaa tcgctagagc caaatgggaa ttcttctatg gctccttgga cccccccagc
     2101 tcaggtgcta agcccccaga gcaggccccc ccatctccac ctggggtggg ctcaaggcag
     2161 ggctctgggg tggctgtggg gcgagcagcc aagtactccg agacagacct ggacacggtg
     2221 cccctgaggt gctaccgcga gactgacatc gatgaggtgc tggctgagcg ggaggaggcc
     2281 gactcggcca tcgaaagtca gcccagctct gagggcccac caggcactgc ctacccacct
     2341 gccccacggc ccggcccact ccctggccct catcccagcc tcggcagtgg caatgaggat
     2401 gaggacgacg atgaggcagg tggggaagaa gatgtggacg acgaggtgtt tgaggcctct
     2461 gaaggggccc ggccagggag ccggatgcct ctcaagtcac ctgtgccctt tctacctggg
     2521 acgagcccct cggctgatgg gcctgactct ttcagttgtg tgttcgaagc catcctggag
     2581 tcacaccggg ccaaaggcac ctcctatacc agcctcgcct cgctggaggc cttggcctca
     2641 cctggcccaa cccagagccc cttcttcacc tttgagctgc ctccccaacc ccctgcaccc
     2701 cggcccgacc caccagctcc cgccccactt gcccctcttg aaccggattc tggtaccagc
     2761 tctgctgctg atggtccttg gacacagaga ggggaggagg aggaggcaga ggccagagcc
     2821 aagctggccc cagggaggga gccccctagt ccctgccact cagaggacag ccttgggctg
     2881 ggggcagcac cccttggcag cgaaccaccc ctgagccagc tggtgtccga ctcagactca
     2941 gagctggaca gcacagagcg gctggccctg ggaagcacag acaccttgtc caatgggcag
     3001 aaagcggacc tggaggctgc gcagcgcctg gccaagaggc tgtaccgact agatggcttc
     3061 aggaaggccg atgtggcccg gcacctgggc aagaacaatg acttcagcaa actggtggct
     3121 ggggagtacc tcaagttctt tgtcttcacg ggcatgactc tggaccaagc tctcagggtg
     3181 tttctgaagg agctggcctt aatgggtgag acccaggaac gagagcgcgt gctggcccac
     3241 ttctcccagc gatacttcca gtgcaatcct gaagccctgt cctcagagga cggcgcccac
     3301 acgctgacct gtgcgctcat gctgctcaac acggatctcc acggccataa catcgggaag
     3361 cgcatgacct gcggggactt catcgggaac ctggagggcc tcaatgatgg cggcgacttc
     3421 cctagggagc tgctcaaggc cttgtacagc tccatcaaga atgagaagct gcagtgggcc
     3481 atagacgagg aggagctgag acgctctctg tctgagttgg ccgaccccaa ccccaaggtc
     3541 atcaagcgga tcagcggggg cagtggcagt ggctccagcc ctttcctgga cctgactccc
     3601 gagcctgggg ctgccgtcta caagcacggg gccctggtgc gaaaggtgca cgcagaccct
     3661 gactgcagga agacacctcg gggcaagcgg ggctggaaga gcttccacgg gatcctcaag
     3721 ggcatgatcc tctacctgca gaaggaggag tacaagcctg ggaaggccct ttcagagacg
     3781 gagctcaaga atgccatcag catccaccat gccctggcca ctcgtgccag tgactacagc
     3841 aagaggcccc acgtcttcta cctgcgcaca gctgactggc gggtcttcct cttccaggcc
     3901 ccgagcctgg agcagatgca gtcctggatc actcgcatca atgtagtagc cgctatgttc
     3961 tctgcgcccc ccttcccagc tgctgttagc tcccaaaaga agttcagccg ccctctcctg
     4021 cccagcgctg ccacccgcct ctcccaggag gagcaggtgc ggacccacga ggccaagctg
     4081 aaggccatgg caagtgagct gcgggagcac cgggccgccc agctgggcaa gaagggccgg
     4141 ggcaaggagg ctgaagagca gcggcagaag gaggcctacc tggagtttga gaaatcccgc
     4201 tacagcacct atgcagcgct gcttcgggtc aagctgaagg caggcagtga ggagctggat
     4261 gcagtggagg cagcactggc ccaggccggg agcacagagg atggactccc tccttctcac
     4321 tccagtccct ccctgcagcc caaaccctcc agccagcccc gggctcagcg tcacagctca
     4381 gagcctcggc caggggcagg cagtgggcgg cggaagccct gagtcgacgg taccgcgggc
     4441 ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
     4501 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     4561 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4621 acaaatttta caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4681 atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     4741 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4801 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4861 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4921 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4981 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     5041 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     5101 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     5161 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5221 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5281 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     5341 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5401 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5461 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     5521 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5581 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5641 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     5701 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     5761 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     5821 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     5881 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5941 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     6001 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     6061 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     6121 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     6181 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     6241 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     6301 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     6361 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     6421 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     6481 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     6541 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     6601 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     6661 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     6721 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     6781 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     6841 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     6901 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     6961 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7021 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7081 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7141 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7201 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7261 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7321 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7381 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7441 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7501 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7561 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7621 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7681 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7741 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
