LOCUS       sequence-410649-        7036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of hEFA6C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7036)
  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 7036)
  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-hEFA6C
FEATURES             Location/Qualifiers
     source          1..7036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      598..607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(604..606,607..609,610..1320)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(604..606,607..609,610..1320)
                     /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(649..670)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(910..929)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1257..1278
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3815..3936
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3852..3871)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3906..3925
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3943..4398)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4080..4101)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4292..4311
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4425..4529
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4531..4888
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4531..4722
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4554..4574)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4739..4874
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4923..5717
                     /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             4923..5717
                     /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(4977..4996)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5587..5606
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5905..5924)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5949..5996
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6325..6913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6814..6833
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc
      601 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      661 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      721 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      781 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
      841 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      901 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      961 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1021 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     1081 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1141 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1201 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1261 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1321 tccggactca gatctaccat ggaggaggac aagctcttat ctgcagtgcc tgaggaaggc
     1381 gatgccaccc gtgaccccgg tccagagcct gaagaggagc caggggtccg gaatgggatg
     1441 gccagtgagg gcctgaacag cagcctctgc agcccagggc acgagcgaag gggcacccca
     1501 gcggacactg aggaacccac gaaggaccca gatgtggcct tccatggcct cagccttggc
     1561 ctctctctca ccaatggcct agccctgggg ccagacttga acattctgga agattcagcg
     1621 gagtccaggc cctggagggc tggcgtgctg gcagaggggg acaatgcttc caggagcctc
     1681 tacccagatg ctgaggaccc tcagctgggg ttggatggtc ccggggagcc agatgtgcgg
     1741 gatggcttca gcgccacgtt tgagaagatt ctggagtcag agctgctgcg gggcacccag
     1801 tacagcagcc tcgactccct agacgggctg agcctcacgg atgagagcga cagctgcgtc
     1861 agcttcgagg cccccctcac acccctcatc cagcagcggg cccgtgacag ccctgagcca
     1921 ggggctgggt tgggcattgg ggacatggcg tttgaggggg acatgggggc ggctggtggt
     1981 gatggggagc tgggcagccc cctgcggcgc tccatctcca gcagccgctc tgagaatgtc
     2041 ctgagccgcc tgtctctcat ggccatgccc aatggattcc atgaagatgg ccctcagggc
     2101 ccaggggggg atgaggatga tgatgaggag gacacggaca agttgctgaa ctcagccagt
     2161 gaccccagcc tgaaggatgg cctgtcagac tcagactctg agctcagcag ctcggagggg
     2221 ttggagcctg gtagtgcaga ccctctggcc aacgggtgcc agggggtcag tgaagctgct
     2281 catcggctgg cacgccgtct ctaccacctc gagggcttcc agcgctgtga tgtggcccgg
     2341 cagctgggca agaacaacga gtttagcagg ctggtggccg gggagtacct cagtttcttc
     2401 gacttctcgg gcttgactct ggacggagca ctcagaacat tcttgaaggc cttcccgctg
     2461 atgggggaga cacaagagcg tgagcgggtc ctcacacact tctcccgccg gtactgccag
     2521 tgcaaccctg atgacagcac ttcggaagat gggatccaca cgctcacctg tgccctgatg
     2581 ctgctcaaca cggacctgca cggccacaac attggcaaaa agatgtcctg tcagcaattc
     2641 attgccaact tggaccagct gaatgatggc caagactttg ccaaagacct gctgaagacc
     2701 ctttacaact ccatcaagaa tgaaaagctg gaatgggcca ttgatgagga tgagctgagg
     2761 aaatccctgt ctgagctggt ggatgacaag ttcgggacag gcacgaagaa ggtgacgcga
     2821 atcctggatg gtggcaaccc cttcctggat gtcccacagg cgctcagtgc caccacctac
     2881 aagcacggcg tcctgacccg gaagactcac gctgacatgg atggcaagag gacgccccgt
     2941 gggaggcgtg gctggaagaa attctacgca gtgctcaaag ggaccatcct gtacctgcag
     3001 aaggatgagt acaggcctga caaagctcta tcggagggtg acctgaagaa cgccattcgc
     3061 gtgcatcacg ctctggccac cagggcctct gactacagca agaagtccaa cgtgctgaag
     3121 cttaagacag ccgactggag ggtattcctc ttccaggcac cgagcaagga agaaatgctg
     3181 tcctggatcc tcaggatcaa cctggtggca gccatcttct ctgccccggc cttcccagcc
     3241 gctgtcagct ccatgaagaa gttctgtcgg cccctgctgc cctcctgcac cacccgcctc
     3301 tgccaggagg agcaactgcg gtctcatgag aataagttga ggcagctgac tgcggagctg
     3361 gccgaacaca ggtgtcaccc agtcgagagg ggcatcaagt ccaaggaggc cgaggagtac
     3421 cggttgaagg agcactatct caccttcgag aaaagccgtt atgagaccta tatccacctc
     3481 ctggctatga aaatcaaagt gggctcagat gatctggagc ggattgaggc ccggctggcc
     3541 actctggaag gggatgaccc ttctctccgg aagacacatt caagccctgc cctcagccag
     3601 ggccatgtga ctggcagcaa aaccacaaag gatgccactg ggcctgatac ttaggaattc
     3661 tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag
     3721 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3781 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3841 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3901 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     3961 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4021 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4081 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4141 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4201 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4261 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4321 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4381 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4441 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4501 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     4561 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4621 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4681 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4741 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4801 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4861 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4921 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4981 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5041 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5101 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5161 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5221 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5281 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5341 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5401 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5461 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5521 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5581 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5641 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5701 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5761 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5821 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5881 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5941 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     6001 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     6061 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     6121 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     6181 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6241 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6301 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6361 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6421 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6481 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6541 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6601 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6661 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6721 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6781 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6841 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6901 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6961 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7021 catgcattag ttatta
//
