LOCUS       sequence-410645-        7881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of hEFA6B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7881)
  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 7881)
  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-hEFA6B
FEATURES             Location/Qualifiers
     source          1..7881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..354
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        355..558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     509..529
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(603..605,606..608,609..1319)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(603..605,606..608,609..1319)
                     /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(648..669)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(909..928)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1256..1277
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4659..4780
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4696..4715)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4750..4769
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4787..5242)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4924..4945)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5136..5155
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5269..5373
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5375..5732
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5375..5566
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5398..5418)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5583..5718
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5767..6561
                     /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             5767..6561
                     /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(5821..5840)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6431..6450
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6749..6768)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6793..6840
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7169..7757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7658..7677
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
       61 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      121 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      181 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      241 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      301 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      361 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      421 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      481 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      541 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca
      601 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      661 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      721 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      781 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
      841 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      901 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      961 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1021 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     1081 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1141 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1201 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1261 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1321 ccggactcag atctaccatg atgggtgact acagactccc tgaccacccc cagcccatgg
     1381 aaattctcaa cctgtacttg ggagacagcc tggagcccca cccaggagag tgcccaaggg
     1441 aaacgtgcag ccatgaggat ccaccggagc ctttcgagga gcaaacctgg gccactgacc
     1501 ctcctgaacc taccagacaa aatgttcctc cctggggctc cggtgtggag ctcacacacc
     1561 tggggagctg ggtccatcag gacgggctgg agccttgcca ggagcaaacc cgggccactg
     1621 accctcctga atctaccaga caagatgctc ctccctgggg ctccggtgtg gagctcacac
     1681 acctggggag cccctctgcc cagagggagc acaggcagaa cacagcatca ccagggtcac
     1741 cagtgaacag ccatctaccg gggagcccaa agcagaaccg gagcacgtcc acacaggtag
     1801 tgttctgggc aggcatcctg caggcccaga tgtgtgtcct agacctggag gaggagctgg
     1861 agaagacgga agggctcaag gctgggctga aatgctgtct ccccacgccc cctgtggacc
     1921 tccccgggga cacgggcctg cactccagcc cacctgagaa tgaagactca ggggaagaca
     1981 gcagcgagcc tgagggagag ggccaggcat ggctgagaga gggaacccca gactcttccc
     2041 cacagtgggg agctgaggag gagagcatgt tcttcagcaa ccccctcttc ctggcgagtc
     2101 cttgctcaga gaacagtgct tctggagagt gcttttcctg gggggcttca gactcccatg
     2161 caggtgtgag gactggacct gagagcccag cgactctgga gcctcccctc ccagaagaca
     2221 cagtgctgtg ggagctggaa agtgagccag atttggggga cggcgctgct atcagtgggc
     2281 attgtacccc tccattccct gtgcccatct ataaaccaca ctccatctgc tgggcctcag
     2341 tggctgccgc tgagggggct cctgcagcac ctcctggtca cggggagagt gagggagata
     2401 ggcttggtcc tgctccatct gcagcaccgt gtgtggacga agcattgacc tgggaatcag
     2461 gatgtgtcgg atctgatctt ggccctgctg cacatcctgt gcaaccttgg gcctctctca
     2521 gccctgaggg ctggcagaga ggaggtcctt tttggcccca ggtgactctt aactcccagg
     2581 acagagatga gagggagggt ggacaccccc aggaatctct tccctgcacc ttggccccct
     2641 gcccctggag gagcccagct tcttctccag agcctagcag cccagaatct gagagcagag
     2701 gccctggtcc caggcccagc cctgcatcgt cccaggaggg cagcccgcag cttcaacacc
     2761 acagctcagg cattttgccc aagtggacac tagatgcttc acagtcttca ctcttggaga
     2821 cggatgggga acagccaagt tccttgaaga aaaaggaggc aggggaggcc ccaaaaccag
     2881 gcgaggaagt aaagagtgaa ggaacagcca ggcctgcaga gactggagac gtccagcctg
     2941 acattcacct gacttctgca gaacatgaga atctgaggac accgatgaac tcttcttggc
     3001 ttcctgggag ccctatgccc caagcacagt ccccagagga aggccagaga ccaccagctg
     3061 gagacaagct agctaatggc gtcaggaaca acaaggtagc ctggaacttg gcctcacgcc
     3121 tctatcgcct ggagggcttc cggaagtctg aagtggctgc ctacctgcag aagaacaatg
     3181 actttagcag ggctgtggct gaggagtacc tgtccttctt ccagtttgga ggccagagtc
     3241 tggaccgagc cctccggagc ttcctccagg ctttggtgct cagtggggag actcaggaac
     3301 gggagcgaat cctctaccag ttctccagac gcttccacca ttgcaatccg gggatcttcc
     3361 cctcagtaga ttctgtacac accttgacat gtgcaatcat gctgcttaac acggacctgc
     3421 atggacagaa cattgggaag agcatgagct gccaggaatt cataaccaac ctgaatgggc
     3481 tgagggatgg cgggaacttc cccaaggagc tgctgaaggc cctctactgg tctatccgca
     3541 gcgagaagct cgagtgggcc gtggatgaag aagacacagc cagacctgag aaggcccagc
     3601 cgtccctgcc agctggcaag atgagcaagc ccttccttca gctggctcag gatcccacag
     3661 tgcccaccta caagcagggc atcctggctc ggaaaatgca tcaagatgca gacggcaaga
     3721 agacgccatg gggcaagcgt ggctggaaga tgttccacac cttactgcga gggatggttc
     3781 tctacttcct gaagcaggga gaagaccact gtctggaggg ggagagcttg gtggggcaga
     3841 tggtggatga gcccgtgggg gtgcaccact cgctggccac ccccgccacg cattacacca
     3901 agaagccgca cgtcttccag ctgcgcacgg ctgactggcg cctctacctc ttccaggcac
     3961 ccactgccaa ggagatgagc tcctggatcg cgcgcatcaa cttggctgcg gccacccact
     4021 ccgcgccgcc cttccccgcc gctgtgggct cccagcgcag attcgtgcgg cccatcctgc
     4081 ccgtgggccc cgcccagagc tccctggagg agcagcatcg atcccacgag aactgcctgg
     4141 acgctgccgc ggacgacctg ctggatctac agaggaacct gccggagcgg cggggccgtg
     4201 gccgcgagct ggaggagcac cgcctgcgga aggagtacct ggagtacgag aaaacccgct
     4261 acgagaccta cgtgcagctg ctggtggccc gcctgcactg cccctctgat gctctggacc
     4321 tgtgggagga gcagctgggg agggaagctg gaggcactcg ggagcccaag ctcagcctga
     4381 agaagtccca ctcgagcccg tccctgcacc aggatgaggc tcccaccacg gccaaggtga
     4441 agcgcaacat ctcagagcgc agaacctacc ggaagatcat ccctaagcgg aaccgcaatc
     4501 agctgtgagt cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     4561 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     4621 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     4681 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4741 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     4801 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4861 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4921 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4981 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     5041 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     5101 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     5161 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     5221 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5281 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5341 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     5401 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5461 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5521 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5581 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5641 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     5701 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     5761 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5821 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5881 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5941 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     6001 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6061 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6121 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6181 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     6241 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     6301 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6361 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6421 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6481 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6541 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     6601 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6661 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6721 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     6781 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     6841 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     6901 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     6961 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     7021 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     7081 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7141 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7201 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7261 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7321 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7381 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7441 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7501 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7561 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7621 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7681 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7741 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7801 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7861 ccgccatgca ttagttatta a
//
