LOCUS       sequence-263664-        5467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of HA-tagged human UFM1 wildtype.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5467)
  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 5467)
  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-HA-UFM1
FEATURES             Location/Qualifiers
     source          1..5467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             914..940
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1242..1260)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1280..1297)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1286..1510
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1556..1984
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1993..2013)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1998..2327
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2178..2313
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2347..2364)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2347..2363)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2356..2378)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2576..3031
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3048..3067)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3167..3189
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3227..3245)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3313..3417
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3418..3486,3487..4278)
                     /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(3636..3655)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4449..5037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4938..4957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5191..5208
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5325..5346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5361..5366,5367..5384,5385..5391)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5399..5415
                     /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     5404..5426
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5423..5439
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5423..5439
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctaa
      901 gcttgccgcc atgtacccat acgatgttcc agattacgct gggtcgacca tgtcgaaggt
      961 ttcctttaag atcacgctga cgtcggaccc acggctgccg tacaaagtac tcagtgttcc
     1021 tgaaagtaca cctttcacag cagtcttaaa gtttgcagca gaagaattta aagttcctgc
     1081 tgcaacaagt gcaattatta ccaatgatgg aataggaata aatcctgcac agactgctgg
     1141 aaatgttttt ctaaaacatg gttcagaact gcggattatt cctagagatc gtgttggaag
     1201 ttgttaagcg gccgctcgag catgcatcta gagggcccta ttctatagtg tcacctaaat
     1261 gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1321 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1381 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1441 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     1501 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     1561 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1621 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1681 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1741 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1801 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1861 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1921 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1981 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2041 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2101 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2161 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2221 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     2281 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctgttaacag
     2341 cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     2401 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2461 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt
     2521 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     2581 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2641 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2701 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2761 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     2821 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2881 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
     2941 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3001 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     3061 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3121 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3181 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     3241 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3301 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3361 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3421 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3481 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3541 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3601 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3661 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3721 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3781 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3841 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3901 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3961 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4021 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4081 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4141 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4201 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4261 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4321 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4381 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4441 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4501 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4561 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4621 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4681 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4741 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4801 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4861 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4921 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4981 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5041 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5101 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     5161 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     5221 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     5281 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     5341 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     5401 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa ttaattcgag
     5461 ctcgccc
//
