LOCUS       sequence-451044-        4230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of yeast Ubc1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4230)
  AUTHORS   Beckwith R, Estrin E, Worden EJ, Martin A
  TITLE     Reconstitution of the 26S proteasome reveals functional asymmetries 
            in its AAA+ unfoldase.
  JOURNAL   Nat Struct Mol Biol. 2013 Oct;20(10):1164-72. doi: 
            10.1038/nsmb.2659. Epub 2013 Sep 8.
  PUBMED    24013205
REFERENCE   2  (bases 1 to 4230)
  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/nsmb.2659";
            journalName: "Nat Struct Mol Biol"; date: "2013-10"; volume: "20"; 
            issue: "10"; pages: "1164-72"
COMMENT     Sequence Label: pAM263
FEATURES             Location/Qualifiers
     source          1..4230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(34..102,103..894)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(252..271)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1065..1653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1554..1573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1807..1824
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1839..1978
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1904..1923)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2064..2086
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2080..2271)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2780..2799
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2780..2798
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2830..2852
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3510..3527
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3580..3598)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3594..3641
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3678..4133
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3765..3784)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3975..3996
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        join(4159..4230,1..33)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
       61 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
      121 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
      181 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
      241 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
      301 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
      361 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
      421 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
      481 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
      541 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
      601 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
      661 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
      721 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
      781 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
      841 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
      901 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
      961 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1021 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1081 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1141 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1201 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1261 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1321 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1381 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1441 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1501 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1561 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1621 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     1681 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1741 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     1801 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     1861 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
     1921 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     1981 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     2041 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     2101 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     2161 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     2221 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     2281 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     2341 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     2401 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     2461 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     2521 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     2581 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     2641 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     2701 ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga
     2761 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
     2821 agaaataatt ttgtttaact ttaagaagga gatataccat gagtagggct aagagaatta
     2881 tgaaagaaat ccaagctgtg aaggatgatc ctgcagctca cattactctt gaatttgtga
     2941 gtgaatctga tatccaccat ttaaaaggca catttttggg cccacctgga acaccttacg
     3001 agggtggcaa atttgtcgtg gatatcgaag tacctatgga gtatccattc aaaccaccaa
     3061 agatgcagtt cgacacaaaa gtataccatc caaatatatc atcagtgaca ggtgccattt
     3121 gtttagatat tcttaagaat gcatggtcgc cagtgataac actaaagtct gcattgattt
     3181 cgctacaggc gctactacaa tctccagagc ctaacgatcc tcaagatgca gaagtagcac
     3241 aacactattt acgggataga gaatctttta ataagactgc tgcactatgg acgaggttat
     3301 acgccagtga aacttccaat ggtcaaaaag gcaatgtaga ggagtctgat ttatatggga
     3361 ttgaccacga tctgattgac gagtttgaat ctcaaggttt tgaaaaggac aagattgtgg
     3421 aagtgttgag acgattaggc gtcaagtcct tagaccccaa tgacaacaac acagccaacc
     3481 gtatcatcga ggaattgttg aagctcgagc accaccacca ccaccactga gatccggctg
     3541 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     3601 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     3661 ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3721 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3781 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3841 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3901 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3961 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4021 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4081 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     4141 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4201 atgtatccgc tcatgagaca ataaccctga
//
