LOCUS       sequence-176221-        5254 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally EGFP-tagged Arf1(WT) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5254)
  AUTHORS   Kondo Y, Hanai A, Nakai W, Katoh Y, Nakayama K, Shin HW
  TITLE     ARF1 and ARF3 are required for the integrity of recycling endosomes 
            and the recycling pathway.
  JOURNAL   Cell Struct Funct. 2012;37(2):141-54. Epub 2012 Sep 5.
  PUBMED    22971977
REFERENCE   2  (bases 1 to 5254)
  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"; journalName: "Cell Struct
            Funct."; date: "2012"; volume: "37(2)"; pages: "141-54. Epub 2012 
            Sep 5"
COMMENT     Sequence Label: pEGFP-N3/hArf1(WT)-EGFP
FEATURES             Location/Qualifiers
     source          1..5254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     570..589
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(888..907)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    932..979
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1308..1896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1797..1816
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2071..2374
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2375..2578
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2529..2549
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2625..2634
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3204..3213
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3210..3212,3213..3215,3216..3929)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3210..3212,3213..3215,3216..3926)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3255..3276)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3516..3535)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3863..3884
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4052..4173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4089..4108)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4143..4162
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4180..4635)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4317..4338)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4529..4548
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4662..4766
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4768..5125
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4768..4959
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4791..4811)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4976..5111
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5160..5254,1..700)
                     /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             join(5160..5254,1..700)
                     /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(5214..5233)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
       61 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
      121 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
      181 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
      241 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
      301 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
      361 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
      421 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
      481 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
      541 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
      601 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
      661 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
      721 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
      781 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
      841 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
      901 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
      961 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     1021 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     1081 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     1141 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     1201 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     1261 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1321 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1381 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1441 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     1501 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     1561 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1621 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     1681 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     1741 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     1801 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     1861 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     1921 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     1981 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     2041 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     2101 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     2161 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     2221 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     2281 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     2341 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     2401 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     2461 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     2521 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     2581 tttagtgaac cgtcagatcc gctagcgcta ccggactcag atccgccacc atggggaaca
     2641 tcttcgccaa cctcttcaag ggcctttttg gcaaaaaaga aatgcgcatc ctcatggtgg
     2701 gcctggatgc tgcagggaag accacgatcc tctacaagct taagctgggt gagatcgtga
     2761 ccaccattcc caccataggc ttcaacgtgg aaaccgtgga gtacaagaac atcagcttca
     2821 ctgtgtggga cgtgggtggc caggacaaga tccggcccct gtggcgccac tacttccaga
     2881 acacacaagg cctgatcttc gtggtggaca gcaatgacag agagcgtgtg aacgaggccc
     2941 gtgaggagct catgaggatg ctggccgagg acgagctccg ggatgctgtc ctcctggtgt
     3001 tcgccaacaa gcaggacctc cccaacgcca tgaatgcggc cgagatcaca gacaagctgg
     3061 ggctgcactc actacgccac aggaactggt acattcaggc cacctgcgcc accagcggcg
     3121 acgggctcta tgaaggactg gactggctgt ccaatcagct ccggaaccag aagctcgacg
     3181 gtaccgcggg cccgggatcc atcgccacca tggtgagcaa gggcgaggag ctgttcaccg
     3241 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3301 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     3361 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     3421 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3481 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     3541 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     3601 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     3661 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     3721 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     3781 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     3841 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3901 ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca
     3961 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     4021 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     4081 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4141 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     4201 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4261 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4321 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4381 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     4441 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4501 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     4561 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     4621 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4681 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4741 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     4801 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4861 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4921 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4981 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5041 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5101 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     5161 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5221 gctatgactg ggcacaacag acaatcggct gctc
//
