LOCUS       sequence-263601-        5428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human wild type UFM1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5428)
  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 5428)
  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 WT
FEATURES             Location/Qualifiers
     source          1..5428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    76..97
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(112..117,118..135,136..142)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    150..166
                     /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     155..177
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     174..190
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     174..190
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        219..598
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        599..802
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     753..773
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     799..823
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1033..1051
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1064..1083
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1421..1439)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1459..1476)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1465..1689
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1735..2163
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2172..2192)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2177..2506
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2357..2492
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2526..2543)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2526..2542)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2535..2557)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2755..3210
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3227..3246)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3346..3368
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3406..3424)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3492..3596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3597..3665,3666..4457)
                     /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(3815..3834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4628..5216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5117..5136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5370..5387
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
       61 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
      121 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      181 cagctatgac atgattacga attaattcga gctcgcccga cattgattat tgactagtta
      241 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      301 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      361 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      421 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      481 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      541 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      601 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      661 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      721 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      781 ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc
      841 acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg
      901 gtgcattgga acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagagtcta
      961 taggcccacc cccttggctt cgttagaacg cggctacaat taatacataa ccttatgtat
     1021 catacacata cgatttaggt gacactatag aataacatcc actttgcctt tctctccaca
     1081 ggtgtccact cccaggtcca actgcacctc ggttctaagc ttgccgccat gtcgaaggtt
     1141 tcctttaaga tcacgctgac gtcggaccca cggctgccgt acaaagtact cagtgttcct
     1201 gaaagtacac ctttcacagc agtcttaaag tttgcagcag aagaatttaa agttcctgct
     1261 gcaacaagtg caattattac caatgatgga ataggaataa atcctgcaca gactgctgga
     1321 aatgtttttc taaaacatgg ttcagaactg cggattattc ctagagatcg tgttggaagt
     1381 tgttaagcgg ccgctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg
     1441 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     1501 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     1561 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     1621 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     1681 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     1741 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     1801 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     1861 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     1921 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     1981 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2041 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2101 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2161 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2221 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2281 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2341 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2401 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     2461 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tgttaacagc
     2521 ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     2581 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     2641 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     2701 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     2761 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2821 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2881 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2941 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     3001 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3061 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     3121 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3181 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     3241 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3301 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3361 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3421 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3481 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3541 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3601 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3661 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3721 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3781 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3841 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3901 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3961 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4021 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4081 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4141 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4201 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4261 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4321 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4381 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4441 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4501 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4561 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4621 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4681 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4741 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4801 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4861 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4921 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4981 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5041 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5101 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5161 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5221 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5281 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5341 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5401 gcccaatacg caaaccgcct ctccccgc
//
