LOCUS       sequence-410871-        5746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of hArfaptin2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5746)
  AUTHORS   Shin HW, Kobayashi H, Kitamura M, Waguri S, Suganuma T, Uchiyama Y, 
            Nakayama K
  TITLE     Roles of ARFRP1 (ADP-ribosylation factor-related protein 1) in 
            post-Golgi membrane trafficking.
  JOURNAL   J Cell Sci. 2005 Sep 1;118(Pt 17):4039-48. doi: 10.1242/jcs.02524.
  PUBMED    16129887
REFERENCE   2  (bases 1 to 5746)
  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: "J Cell 
            Sci."; date: "2005-09-1"; volume: "118(Pt 17):4039-48. doi"; pages: 
            " 10.1242/jcs.02524"
COMMENT     Sequence Label: pEGFP-C1-hArfaptin2
FEATURES             Location/Qualifiers
     source          1..5746
                     /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    2534..2655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2571..2590)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2625..2644
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2662..3117)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2799..2820)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3011..3030
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3144..3248
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3250..3607
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3250..3441
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3273..3293)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3458..3593
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3642..4436
                     /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             3642..4436
                     /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(3696..3715)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4306..4325
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4624..4643)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4668..4715
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5044..5632
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5533..5552
                     /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 acggacggga tcctagggaa ggcagccaca
     1381 atggagatcc ctatccacgg gaacggcgaa gccaggcagc ttcctgaaga tgatgggctg
     1441 gagcaggacc tccagcaggt gatggtgtca ggacccaacc tcaatgaaac cagcattgtg
     1501 tctggtggct atgggggctc tggtgatgga ctcatcccca cagggtctgg ccgccatcca
     1561 tctcacagca ccactccttc tggccctgga gatgaggtgg ctcggggcat tgctggagaa
     1621 aagtttgaca tcgtcaagaa atggggcatc aacacctata agtgcacaaa gcaactgtta
     1681 tcagaacgat ttggtcgagg ctcacggact gtggacctgg agctagagct gcagattgag
     1741 ttgctgcgtg agacgaagcg caagtatgag agtgtcctgc agctgggccg ggcactgaca
     1801 gcccacctct acagcctgct gcagacccag catgcactgg gtgatgcctt tgctgacctc
     1861 agccagaagt ccccagagct tcaggaggaa tttggctaca atgcagagac acagaaacta
     1921 ctatgcaaga atggggaaac gctgctagga gccgtgaact tctttgtctc tagcatcaac
     1981 acattggtca ccaagaccat ggaagacacg ctcatgactg tgaaacagta tgaggctgcc
     2041 aggctggaat atgatgccta ccgaacagac ttagaggagc tgagtctagg cccccgggat
     2101 gcagggacac gtggtcgact tgagagtgcc caggccactt tccaggccca tcgggacaag
     2161 tatgagaagc tgcggggaga tgtggccatc aagctcaagt tcctggaaga aaacaagatc
     2221 aaggtgatgc acaagcagct gctgctcttc cacaatgctg tgtccgccta ctttgctggg
     2281 aaccagaaac agctggagca gaccctgcag cagttcaaca tcaagctgcg gcctccagga
     2341 gctgagaaac cctcctggct agaggagcag tgagaattct gcagtcgacg gtaccgcggg
     2401 cccgggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     2461 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2521 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2581 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2641 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2701 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2761 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2821 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2881 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2941 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3001 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3061 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3121 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3181 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3241 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3301 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3361 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3421 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3481 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3541 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3601 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3661 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3721 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3781 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     3841 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3901 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3961 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4021 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4081 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4141 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     4201 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4261 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4321 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4381 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4441 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4501 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4561 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4621 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4681 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4741 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4801 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4861 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     4921 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4981 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5041 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5101 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5161 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5221 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5281 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5341 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5401 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5461 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5521 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5581 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5641 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5701 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
