LOCUS       sequence-263634-        5419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human UFM1 mutant lacking the last three C-terminal 
            residues.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5419)
  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 5419)
  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-Ufm1 ∆C3
FEATURES             Location/Qualifiers
     source          1..5419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    66..87
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(102..107,108..125,126..132)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    140..156
                     /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     145..167
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     164..180
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     164..180
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        209..588
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        589..792
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     743..763
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     789..813
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1023..1041
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1054..1073
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1402..1420)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1440..1457)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1446..1670
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1716..2144
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2153..2173)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2158..2487
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2338..2473
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2507..2524)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2507..2523)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2516..2538)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2736..3191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3208..3227)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3327..3349
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3387..3405)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3473..3577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3578..3646,3647..4438)
                     /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(3796..3815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4609..5197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5098..5117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5351..5368
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
       61 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
      121 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
      181 atgattacga attaattcga gctcgcccga cattgattat tgactagtta ttaatagtaa
      241 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      301 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      361 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      421 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      481 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      541 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      601 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      661 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      721 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      781 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
      841 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga
      901 acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagagtcta taggcccacc
      961 cccttggctt cgttagaacg cggctacaat taatacataa ccttatgtat catacacata
     1021 cgatttaggt gacactatag aataacatcc actttgcctt tctctccaca ggtgtccact
     1081 cccaggtcca actgcacctc ggttctaagc ttgccgccat gtcgaaggtt tcctttaaga
     1141 tcacgctgac gtcggaccca cggctgccgt acaaagtact cagtgttcct gaaagtacac
     1201 ctttcacagc agtcttaaag tttgcagcag aagaatttaa agttcctgct gcaacaagtg
     1261 caattattac caatgatgga ataggaataa atcctgcaca gactgctgga aatgtttttc
     1321 taaaacatgg ttcagaactg cggattattc ctagagatcg tgtttaagcg gccgctcgag
     1381 catgcatcta gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc
     1441 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     1501 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     1561 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     1621 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     1681 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     1741 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     1801 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     1861 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     1921 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     1981 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2041 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2101 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     2161 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2221 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2281 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2341 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2401 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2461 ttttttggag gcctaggctt ttgcaaaaag ctgttaacag cttggcactg gccgtcgttt
     2521 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     2581 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     2641 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     2701 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     2761 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2821 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2881 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2941 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3001 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3061 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta
     3121 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3181 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3241 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3301 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3361 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3421 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3481 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3541 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3601 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3661 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3721 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3781 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3841 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3901 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3961 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4021 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4081 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4141 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4201 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4261 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4321 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4381 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4441 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4501 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4561 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4621 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4681 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4741 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4801 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4861 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4921 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4981 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5041 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5101 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5161 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5221 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5281 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5341 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     5401 tctccccgcg cgttggccg
//
