LOCUS       sequence-263666-        7249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FLAG-tagged human UFM1 wildtype.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7249)
  AUTHORS   Wang L, Xu Y, Rogers H, Saidi L, Noguchi CT, Li H, Yewdell JW, 
            Guydosh NR, Ye Y
  TITLE     UFMylation of RPL26 links translocation-associated quality control 
            to endoplasmic reticulum protein homeostasis.
  JOURNAL   Cell Res. 2020 Jan;30(1):5-20. doi: 10.1038/s41422-019-0236-6. Epub 
            2019 Oct 8.
  PUBMED    31595041
REFERENCE   2  (bases 1 to 7249)
  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: 
            "10.1038/s41422-019-0236-6"; journalName: "Cell Res"; date: 
            "2020-01"; volume: "30"; issue: "1"; pages: "5-20"
COMMENT     Sequence Label: pRK5-FLAG-UFM1
FEATURES             Location/Qualifiers
     source          1..7249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        435..453
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     466..485
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      525..534
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             534..557
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(859..877)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(897..914)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    903..1127
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1173..1601
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1615..1944
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1795..1930
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2011..2805
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2065..2084)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2675..2694
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2979..3100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      3156..3165
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3169..3303
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3372..3729
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3372..3563
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      3580..3715
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3749..3766)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3749..3765)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3758..3780)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3978..4433
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4450..4469)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4569..4591
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4629..4647)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4715..4819
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4820..4888,4889..5680)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5038..5057)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5851..6439
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6340..6359
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6593..6610
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6727..6748
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6763..6768,6769..6786,6787..6793)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6801..6817
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6806..6828
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6825..6841
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6825..6841
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6870..7249
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccgggaa
      301 cggtgcattg gaacgcggat tccccgtgcc aagagtgacg taagtaccgc ctatagagtc
      361 tataggccca cccccttggc ttcgttagaa cgcggctaca attaatacat aaccttatgt
      421 atcatacaca tacgatttag gtgacactat agaataacat ccactttgcc tttctctcca
      481 caggtgtcca ctcccaggtc caactgcacc tcggttctaa gcttgccgcc atggactaca
      541 aggacgacga tgacaagggg tcgaccatgt cgaaggtttc ctttaagatc acgctgacgt
      601 cggacccacg gctgccgtac aaagtactca gtgttcctga aagtacacct ttcacagcag
      661 tcttaaagtt tgcagcagaa gaatttaaag ttcctgctgc aacaagtgca attattacca
      721 atgatggaat aggaataaat cctgcacaga ctgctggaaa tgtttttcta aaacatggtt
      781 cagaactgcg gattattcct agagatcgtg ttggaagttg ttaagcggcc gctcgagcat
      841 gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc
      901 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
      961 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     1021 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     1081 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     1141 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     1201 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     1261 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     1321 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     1381 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     1441 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     1501 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     1561 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     1621 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     1681 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     1741 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     1801 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1861 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     1921 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     1981 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     2041 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     2101 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     2161 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     2221 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     2281 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     2341 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     2401 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     2461 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     2521 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     2581 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     2641 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     2701 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     2761 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     2821 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     2881 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     2941 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     3001 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3061 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     3121 ggatccccgg gtaccgagct cgaattcaat cgatggccgc catggcccaa cttgtttatt
     3181 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3241 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     3301 atcgggaatt aattcggcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     3361 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3421 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3481 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3541 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3601 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3661 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3721 ttttgcaaaa agctgttaac agcttggcac tggccgtcgt tttacaacgt cgtgactggg
     3781 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     3841 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     3901 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3961 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4021 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4081 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4141 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     4201 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4261 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg
     4321 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4381 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     4441 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     4501 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4561 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4621 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4681 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4741 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4801 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4861 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4921 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4981 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5041 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5101 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5161 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5221 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5281 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5341 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5401 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5461 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5521 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5581 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5641 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5701 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5761 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5821 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5881 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5941 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6001 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6061 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6121 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6181 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6241 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6301 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6361 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6421 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6481 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6541 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6601 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6661 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6721 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6781 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6841 catgattacg aattaattcg agctcgcccg acattgatta ttgactagtt attaatagta
     6901 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     6961 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     7021 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     7081 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     7141 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     7201 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatg
//
