LOCUS       sequence-328931-        6226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of p58 (ie ERGIC-53; LMAN1) tagged with GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6226)
  AUTHORS   Ward TH, Polishchuk RS, Caplan S, Hirschberg K, Lippincott-Schwartz 
            J
  TITLE     Maintenance of Golgi structure and function depends on the integrity
            of ER export.
  JOURNAL   J Cell Biol. 2001 Nov 12;155(4):557-70. doi: 10.1083/jcb.200107045. 
            Epub 2001 Nov 12.
  PUBMED    11706049
REFERENCE   2  (bases 1 to 6226)
  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: 
            "10.1083/jcb.200107045"; journalName: "J Cell Biol"; date: 
            "2001-11-12- 12"; volume: "155"; issue: "4"; pages: "557-70"
COMMENT     Sequence Label: GFP-p58
FEATURES             Location/Qualifiers
     source          1..6226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        102..405
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        406..609
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     560..580
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     sig_peptide     647..700
                     /codon_start=1
                     /product="signal sequence of chicken lysozyme"
                     /label=lysozyme signal sequence
                     /translation="MRSLLILVLCFLPLAALG"
     CDS             join(704..706,707..1417)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(704..706,707..1417)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(746..767)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1007..1026)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1354..1375
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3055..3176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3092..3111)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3146..3165
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3183..3638)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3320..3341)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3532..3551
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3665..3769
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3771..4128
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3771..3962
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3794..3814)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3979..4114
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4163..4957
                     /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             4163..4957
                     /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(4217..4236)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4827..4846
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5145..5164)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5189..5236
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5565..6153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6054..6073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca
       61 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      121 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      181 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      241 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      301 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      361 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      421 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      481 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      541 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      601 agcagagctg gtttagtgaa ccgtcagatc cgctagcgcc gccaccatga ggtctttgct
      661 aatcttggtg ctttgcttcc tgcccctggc tgctctgggg aaagtgagca agggcgagga
      721 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      781 gttcagcgtg tccggcgagg gcgagggtga tgccacctac ggcaagctga ccctgaagtt
      841 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
      901 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      961 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1021 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1081 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1141 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1201 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1261 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     1321 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1381 cgccgggatc actctcggca tggacgagct gtacaagtac tcaggagggg gtggcgacgg
     1441 catgggagga gatgctgcgg ctcccggcgc cgcaggcacc caggccgagt tgccacaccg
     1501 ccgtttcgag tacaaataca gctttaaggg gccgcatctg gtgcagagcg atgggaccgt
     1561 gcccttttgg gcccacgcgg ggaatgctat tccaagtgca gatcagatcc gaatagcacc
     1621 gtccttaaag agccaaagag gttccgtgtg gaccaaaaca aaagcagcct tcgagaactg
     1681 ggaagtggaa gtgacattcc gagtgaccgg aagaggtcga attggagctg atggcttagc
     1741 aatttggtat acagaaaacc aaggcttgga cggccctgtg tttggatcag ctgatatgtg
     1801 gaatggtgtt ggaatatttt ttgattcttt tgacaatgac ggaaagaaaa acaatcctgc
     1861 tatagtagtt gtaggcaaca atggacaaat caattatgac caccaaaatg atggtgccac
     1921 tcaagcctta gcgagttgtc agagggactt ccgtaataaa ccctatcctg tccgggcaaa
     1981 gattacctat taccagaaaa cactgacggt catgatcaat aatggcttta caccagataa
     2041 aaatgattat gaattctgtg ccaaagtgga aaatatggtt atccccacgc aagggcattt
     2101 tgggatatct gctgcaacag gaggtcttgc agatgaccac gatgttctct ctttcctgac
     2161 tttccaacta accgagcccg gaaaagagcc acctacgcca gaaaaagaca tttcagaaaa
     2221 agaaaaggaa aagtatcaag aggagtttga acacttccag caagaattag acaagaagaa
     2281 ggaggaattc cagaagggcc accctgacct ccaggggcag ccagcggatg acatctttga
     2341 gagcataggg gatcgggagc tgagacaagt gtttgaagga caaaatcgca ttcatctgga
     2401 gatcaagcaa cttaaccggc agctggatat gattctcgat gaacagagga gatacgtgtc
     2461 ttctctgaca gaggagattt ccaggagagg agcagggacc ccaggacagc ctgggcaggt
     2521 ctcccaacag gaactggaca ctgttgtgag gacccagcag gagattttga gacaagtgaa
     2581 tgagatgaag aactccatga gtgagaccat gcggctggtt agtggcgtcc agcacccagg
     2641 ctcggccggc gtctacgaga caacccagca cttcatggac atcaaggagc acctgcacat
     2701 cgtgaagaga gatatcgaca gcctcgcgca gcgcagcatg tcatctaatg aaaaaccaaa
     2761 atgcccagat ctaccagcat tcccgtcgtg tctgtccaca gtccactttg tgatatttat
     2821 agtggtgcag acggtgttgt tcattggcta catcatgtac aggactcagc aagaagcagc
     2881 tgccaaaaaa ttcttctgac tgcagtcgac ggtaccgcgg gcccgggatc caccggatct
     2941 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3001 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3061 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3121 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     3181 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3241 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3301 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3361 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3421 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3481 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3541 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3601 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3661 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3721 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3781 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3841 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3901 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3961 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4021 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4081 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4141 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4201 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4261 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4321 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4381 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4441 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4501 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4561 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4621 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4681 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4741 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4801 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4861 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4921 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4981 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5041 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5101 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5161 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5221 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5281 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5341 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5401 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5461 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5521 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5581 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5641 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5701 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5761 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5821 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5881 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5941 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6001 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6061 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6121 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6181 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgt
//
