LOCUS       sequence-329544-        6409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of p58 (ie ERGIC-53; LMAN1) tagged with 
            HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6409)
  AUTHORS   Weigel AV, Chang CL, Shtengel G, Xu CS, Hoffman DP, Freeman M, Iyer 
            N, Aaron J, Khuon S, Bogovic J, Qiu W, Hess HF, Lippincott-Schwartz 
            J
  TITLE     ER-to-Golgi protein delivery through an interwoven, tubular network 
            extending from ER.
  JOURNAL   Cell. 2021 Apr 29;184(9):2412-2429.e16. doi: 
            10.1016/j.cell.2021.03.035. Epub 2021 Apr 13.
  PUBMED    33852913
REFERENCE   2  (bases 1 to 6409)
  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.cell.2021.03.035"; journalName: "Cell"; date: 
            "2021-04-29- 29"; volume: "184"; issue: "9"; pages: "2412-2429.e16"
COMMENT     Sequence Label: Halo-p58
FEATURES             Location/Qualifiers
     source          1..6409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        40..343
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        344..547
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     498..518
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     sig_peptide     585..638
                     /codon_start=1
                     /product="signal sequence of chicken lysozyme"
                     /label=lysozyme signal sequence
                     /translation="MRSLLILVLCFLPLAALG"
     CDS             642..1529
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     polyA_signal    3176..3297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3213..3232)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3267..3286
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3304..3759)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3441..3462)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3653..3672
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3786..3890
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3892..4249
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3892..4083
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3915..3935)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4100..4235
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4284..5078
                     /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             4284..5078
                     /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(4338..4357)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4948..4967
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5266..5285)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5310..5357
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5686..6274
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6175..6194
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
       61 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      121 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      181 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      241 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      301 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      361 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      421 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      481 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      541 cagagctggt ttagtgaacc gtcagatccg ctagcgccgc caccatgagg tctttgctaa
      601 tcttggtgct ttgcttcctg cccctggctg ctctggggaa agcagaaatc ggtactggct
      661 ttccattcga cccccattat gtggaagtcc tgggcgagcg catgcactac gtcgatgttg
      721 gtccgcgcga tggcacccct gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt
      781 ggcgcaacat catcccgcat gttgcaccga cccatcgctg cattgctcca gacctgatcg
      841 gtatgggcaa atccgacaaa ccagacctgg gttatttctt cgacgaccac gtccgcttca
      901 tggatgcctt catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt cacgactggg
      961 gctccgctct gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat
     1021 ttatggagtt catccgccct atcccgacct gggacgaatg gccagaattt gcccgcgaga
     1081 ccttccaggc cttccgcacc accgacgtcg gccgcaagct gatcatcgat cagaacgttt
     1141 ttatcgaggg tacgctgccg atgggtgtcg tccgcccgct gactgaagtc gagatggacc
     1201 attaccgcga gccgttcctg aatcctgttg accgcgagcc actgtggcgc ttcccaaacg
     1261 agctgccaat cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa tacatggact
     1321 ggctgcacca gtcccctgtc ccgaagctgc tgttctgggg caccccaggc gttctgatcc
     1381 caccggccga agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct gtggacatcg
     1441 gcccgggtct gaatctgctg caagaagaca acccggacct gatcggcagc gagatcgcgc
     1501 gctggctgtc tactctggag atttccggca tgtacaagta ctcaggaggg ggtggcgacg
     1561 gcatgggagg agatgctgcg gctcccggcg ccgcaggcac ccaggccgag ttgccacacc
     1621 gccgtttcga gtacaaatac agctttaagg ggccgcatct ggtgcagagc gatgggaccg
     1681 tgcccttttg ggcccacgcg gggaatgcta ttccaagtgc agatcagatc cgaatagcac
     1741 cgtccttaaa gagccaaaga ggttccgtgt ggaccaaaac aaaagcagcc ttcgagaact
     1801 gggaagtgga agtgacattc cgagtgaccg gaagaggtcg aattggagct gatggcttag
     1861 caatttggta tacagaaaac caaggcttgg acggccctgt gtttggatca gctgatatgt
     1921 ggaatggtgt tggaatattt tttgattctt ttgacaatga cggaaagaaa aacaatcctg
     1981 ctatagtagt tgtaggcaac aatggacaaa tcaattatga ccaccaaaat gatggtgcca
     2041 ctcaagcctt agcgagttgt cagagggact tccgtaataa accctatcct gtccgggcaa
     2101 agattaccta ttaccagaaa acactgacgg tcatgatcaa taatggcttt acaccagata
     2161 aaaatgatta tgaattctgt gccaaagtgg aaaatatggt tatccccacg caagggcatt
     2221 ttgggatatc tgctgcaaca ggaggtcttg cagatgacca cgatgttctc tctttcctga
     2281 ctttccaact aaccgagccc ggaaaagagc cacctacgcc agaaaaagac atttcagaaa
     2341 aagaaaagga aaagtatcaa gaggagtttg aacacttcca gcaagaatta gacaagaaga
     2401 aggaggaatt ccagaagggc caccctgacc tccaggggca gccagcggat gacatctttg
     2461 agagcatagg ggatcgggag ctgagacaag tgtttgaagg acaaaatcgc attcatctgg
     2521 agatcaagca acttaaccgg cagctggata tgattctcga tgaacagagg agatacgtgt
     2581 cttctctgac agaggagatt tccaggagag gagcagggac cccaggacag cctgggcagg
     2641 tctcccaaca ggaactggac actgttgtga ggacccagca ggagattttg agacaagtga
     2701 atgagatgaa gaactccatg agtgagacca tgcggctggt tagtggcgtc cagcacccag
     2761 gctcggccgg cgtctacgag acaacccagc acttcatgga catcaaggag cacctgcaca
     2821 tcgtgaagag agatatcgac agcctcgcgc agcgcagcat gtcatctaat gaaaaaccaa
     2881 aatgcccaga tctaccagca ttcccgtcgt gtctgtccac agtccacttt gtgatattta
     2941 tagtggtgca gacggtgttg ttcattggct acatcatgta caggactcag caagaagcag
     3001 ctgccaaaaa attcttctga ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc
     3061 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3121 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3181 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3241 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     3301 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3361 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3421 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3481 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3541 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3601 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3661 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3721 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3781 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3841 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     3901 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3961 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4021 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4081 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4141 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4201 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4261 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4321 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4381 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4441 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4501 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4561 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4621 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4681 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4741 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4801 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4861 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4921 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4981 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5041 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5101 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5161 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5221 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5281 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5341 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5401 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5461 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5521 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5581 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5641 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5701 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5761 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5821 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5881 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5941 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6001 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6061 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6121 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6181 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6241 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6301 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6361 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaat
//
