LOCUS       sequence-389120-        8100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression of FLAG-UBA6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8100)
  AUTHORS   Hunkeler M, Jin CY, Fischer ES
  TITLE     Structures of BIRC6-client complexes provide a mechanism of 
            SMAC-mediated release of caspases.
  JOURNAL   Science. 2023 Mar 17;379(6637):1105-1111. doi: 
            10.1126/science.ade5750. Epub 2023 Feb 9.
  PUBMED    36758104
REFERENCE   2  (bases 1 to 8100)
  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: 
            "10.1126/science.ade5750"; journalName: "Science"; date: 
            "2023-03-17- 17"; volume: "379"; issue: "6637"; pages: "1105-1111"
COMMENT     Sequence Label: pDARMO_CMVT_N_FLAG_UBA6
FEATURES             Location/Qualifiers
     source          1..8100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      93..102
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             105..128
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             147..170
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3427..3447)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3845..3856)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3923..3940)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3929..4136
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        join(4184..4189,4190..4206,4207..4212)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4382..4915
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  4982..5205
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     rep_origin      complement(5269..5857)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5349..5368)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5896..6687,6688..6756))
                     /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     6519..6538
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6757..6828)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  6853..7018
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     enhancer        7112..7415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7416..7619
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7570..7590
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gagtgacaat gacatccact ttgcctttct ctccacaggt gtccactccc aggtccaaag
       61 ttggccaggc ctagttggcc aggcctagtg ccgccaccat gggtgattac aaggatgacg
      121 atgacaaggg cggtggtagc ggtgggctgg aagttctgtt ccaggggccc ggatccatgg
      181 aaggatccga gcctgtggcc gcccatcagg gggaagaggc gtcctgttct tcctggggga
      241 ctggcagcac aaataaaaat ttgcccatta tgtcaacagc atctgtggaa atcgatgatg
      301 cattgtatag tcgacagagg tacgttcttg gagacacagc aatgcagaag atggccaagt
      361 cccatgtttt cttaagtggg atgggtggtc ttggtttgga aattgcaaag aatcttgttc
      421 ttgcagggat taaggcagtt acaattcatg atacagaaaa atgccaagca tgggatctag
      481 gaaccaactt ctttctcagt gaagatgatg ttgttaataa gagaaacagg gctgaagctg
      541 tacttaaaca tattgcagaa ctaaatccat acgttcatgt cacatcatct tctgttcctt
      601 tcaatgagac cacagatctc tcctttttag ataaatacca gtgtgtagta ttgactgaga
      661 tgaaacttcc attgcagaag aagatcaatg acttttgccg ttctcagtgc cctccaatta
      721 agtttatcag tgcagatgta catggaattt ggtcaaggtt attttgtgat ttcggtgatg
      781 aatttgaagt tttagataca acaggagaag aaccaaaaga aattttcatt tcaaacataa
      841 cgcaagcaaa tcctggcatt gttacttgcc ttgaaaatca tcctcacaaa ctggagacag
      901 gacaattcct aacatttcga gaaattaatg gaatgacagg tttaaatgga tctatacaac
      961 aaataacggt gatatcgcca ttttctttta gtattggtga caccacagaa ctggaaccat
     1021 atttacatgg aggcatagct gtccaagtta agactcctaa aacagttttt tttgaatcac
     1081 tggagaggca gttaaaacat ccaaagtgcc ttattgtgga ttttagcaac cctgaggcac
     1141 ctttagagat tcacacagct atgcttgcct tggaccagtt tcaggagaaa tacagtcgca
     1201 agccaaatgt tggatgccaa caagattcag aagaactgtt gaaactagca acatctataa
     1261 gtgaaacctt ggaagagaag cctgatgtaa atgctgacat tgtgcattgg ctctcttgga
     1321 ctgcccaagg ctttttatct ccacttgctg cagcagtagg aggtgttgcc agccaagaag
     1381 tattgaaagc tgtaacagga aaattttctc ctttgtgcca gtggttatat cttgaagcag
     1441 cagatattgt tgaatcacta ggcaaacctg aatgtgaaga atttctccca cgaggagata
     1501 gatatgatgc cttaagagct tgcattggag acactttgtg tcagaaactg caaaatttaa
     1561 acatcttctt agtagggtgt ggagccatag gctgtgaaat gttgaaaaat tttgctttac
     1621 ttggtgttgg cacaagcaaa gagaaaggaa tgattacagt tacagatcct gacttgatag
     1681 agaaatccaa cttaaataga cagttcctat ttcgtcctca tcacatacag aaacctaaaa
     1741 gctacactgc tgctgatgct actctgaaaa taaattctca aataaagata gatgcacacc
     1801 tgaacaaagt atgtccaacc actgagacca tttacaatga tgagttctat actaaacaag
     1861 atgtaattat tacagcatta gataatgtgg aagccaggag atacgtagac agtcgttgct
     1921 tagcaaatct aaggcctctt ttagattctg gaacaatggg cactaaggga cacactgaag
     1981 ttattgtacc gcatttgact gagtcttaca atagtcatcg ggatccccca gaagaggaaa
     2041 taccattttg tactctaaaa tcctttccag ctgctattga acataccata cagtgggcaa
     2101 gagataagtt tgaaagttcc ttttcccaca aaccttcatt gtttaacaaa ttttggcaaa
     2161 cctattcatc tgcagaagaa gtcttacaga agatacagag tggacacagt ttagaaggct
     2221 gttttcaagt tataaagtta cttagcagaa gacctagaaa ttggtcccag tgtgtagaat
     2281 tagcaagatt aaagtttgaa aaatatttta accataaggc tcttcagctt cttcactgtt
     2341 tccctctgga catacgatta aaagatggca gtttattttg gcagtcacca aagaggccac
     2401 cctctccaat aaaatttgat ttaaatgagc ctttgcacct cagtttcctt cagaatgctg
     2461 caaaactata tgctacagta tattgtattc catttgcaga agaggactta tcagcagatg
     2521 ccctcttgaa tattctttca gaagtaaaga ttcaggaatt caagccttcc aataaggttg
     2581 ttcaaacaga tgaaactgca aggaaaccag accatgttcc tattagcagt gaagatgaga
     2641 ggaatgcaat tttccaacta gaaaaggcta ttttatctaa tgaagccacc aaaagtgacc
     2701 ttcagatggc agtgctttca tttgaaaaag atgatgatca taatggacac atagatttca
     2761 tcacagctgc atcaaatctt cgtgccaaaa tgtacagcat tgaaccagct gaccgtttca
     2821 aaacaaagcg catagctggt aaaattatac ctgctatagc aacaaccact gctacagttt
     2881 ctggcttggt tgccttggag atgatcaaag taactggtgg ctatccattt gaagcttaca
     2941 aaaattgttt tcttaactta gccattccaa ttgtagtatt tacagagaca actgaagtaa
     3001 ggaaaactaa aatcagaaat ggaatatcat ttacaatttg ggatcgatgg accgtacatg
     3061 gaaaagaaga tttcaccctc ttggatttca taaatgcagt caaagagaag tatggaattg
     3121 agccaacaat ggtggtacag ggagtcaaaa tgctttatgt tcctgtaatg cctggtcatg
     3181 caaaaagatt gaagttaaca atgcataaac ttgtaaaacc tactactgaa aagaaatatg
     3241 tggatcttac tgtgtcattt gctccagaca ttgatggaga tgaagatttg ccgggacctc
     3301 cagtaagata ctacttcagt catgacactg attgatgata atggcctgtg gccgtaatgg
     3361 cctgtggccg taaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3421 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3481 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3541 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3601 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3661 tttccccctc cctattgcca cggcggaact cagtgccgcc tgccttgccc gctgctggac
     3721 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     3781 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3841 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3901 tcttccgcgg cttcgagatc tgcctcgact gtgccttcta gttgccagcc atctgttgtt
     3961 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4021 taaaatgagg aaattgcatc aaattgtctg agtaggtgtc attctattct ggggggtggg
     4081 gtggggcagg acagcaaggg ggaggattgg gcagacaata gcaggcatgc tggggactcg
     4141 agttaagggc gaatcgctgg gacgcccgaa cggtggctac tcgttgacat aagcctgttc
     4201 ggttcgtaaa ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac
     4261 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
     4321 tgtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     4381 gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttagg
     4441 tggctcaagt atgggcatca ttcgtacatg taggctcggc cctgaccaag tcaaatccat
     4501 gcgggctgct cttgatcttt tcggtcgtga gttcggtgat gtagccacct actcccaaca
     4561 tcagccggac tccgattacc tcgggaactt gctccgtagt aagacattta ttgcgcttgc
     4621 tgccttcgac caagaagcgg ttgttggcgc tctcgcggct tacgttctgc ccaagtttga
     4681 gcagccgcgt agtgagatct atatctatga tctcgcagtc tccggcgagc accggaggca
     4741 gggcattgcc accgcgctca tcaatctcct caagcatgag gccaacgcgc ttggtgctta
     4801 tgtgatctac gtgcaagcag attacggtga cgatcccgca gtggctctct atacaaagtt
     4861 gggcatacgg gaagaagtga tgcactttga tatcgaccca agtaccgcca cctaacaatt
     4921 cgttcaagcc gagatcggct tcccggccgc ggagttgttc ggtaaattgt cacaacgccg
     4981 cgaatatagt ctttacatgc ccttggccac gcccctcttt aatacgacgg gcaatttgca
     5041 cttcagaaaa tgaagagttt gctttagcca taacaaaagt ccagtatgct ttttcacagc
     5101 ataactggac tgatttcagt ttacaactat tctgtctagt ttaagacttt attgtcatag
     5161 tttagatcta ttttgttcag tttaagactt tattgtccgc ccacacccgc ttacgcaggg
     5221 catctgcgca tccacaggaa gccgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5281 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5341 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5401 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5461 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5521 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5581 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5641 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5701 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5761 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5821 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5881 ggggtctgac gcttattacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5941 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6001 cttaccatct ggccccagtg ctgcaatgat accgcgtgac ccacgctcac cggctccaga
     6061 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6121 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6181 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6241 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6301 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6361 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6421 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6481 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6541 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6601 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6661 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6721 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6781 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttaggaccc
     6841 aagtcaaccg gttgtgggcg gacaaaatag ttgggaactg ggaggggtgg aaatggagtt
     6901 tttaaggatt atttagggaa gagtgacaaa atagatggga actgggtgta gcgtcgtaag
     6961 ctaatacgaa aattaaaaat gacaaaatag tttggaacta gatttcactt atctggttca
     7021 gcctgcggtc cggattcgcc cttaagctag ctagttatta atagtaatca attacggggt
     7081 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     7141 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     7201 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     7261 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     7321 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     7381 agtacatcta cgtattagtc attgctatta ccatggtgat gcggttttgg cagtacatca
     7441 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     7501 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     7561 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     7621 gtttagtgaa ccgtcagatc ctcactctct tccgcctcgc tgtctgcgag ggccagctgt
     7681 tgggctcgcg gttgaggaca aactcttcgc ggtctttcca gtactcttgg atcggaaacc
     7741 cgtcggcctc cgaacggtac tccgccaccg agggacctga gcgagtccgc atcgaccgga
     7801 tcggaaaacc tctcgagaaa ggcgtctaac cagtcacagt cgcaaggtag gctgagcacc
     7861 gtggcgggcg gcagcgggtg gcggtcgggg ttgtttctgg cggaggtgct gctgatgatg
     7921 taattaaagt aggcggtgtt gacaccgcgg atggtcgagg tgaggtgtgg caggcttgag
     7981 atccagctgt tggggtgagt actccctctc aaaagcgggc attacttctg cgctaagatt
     8041 gtcagtttcc aaaaacgagg aggatttgat attcacctgg cccgatctgg ccatacactt
//
