LOCUS       sequence-235389-        5054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the His tagged Saccharomyces cerevisiae Ornithine 
            Decarboxylase in Escherichia coli BL21.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5054)
  AUTHORS   Olin-Sandoval V, Yu JSL, Miller-Fleming L, Alam MT, Kamrad S, 
            Correia-Melo C, Haas R, Segal J, Pena Navarro DA, Herrera-Dominguez 
            L, Mendez-Lucio O, Vowinckel J, Mulleder M, Ralser M
  TITLE     Lysine harvesting is an antioxidant strategy and triggers 
            underground polyamine metabolism.
  JOURNAL   Nature. 2019 Aug;572(7768):249-253. doi: 10.1038/s41586-019-1442-6. 
            Epub 2019 Jul 31.
  PUBMED    31367038
REFERENCE   2  (bases 1 to 5054)
  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/s41586-019-1442-6"; journalName: "Nature"; date: "2019-08";
            volume: "572"; issue: "7768"; pages: "249-253"
COMMENT     Sequence Label: ScSpe1 pET-20b(+)
FEATURES             Location/Qualifiers
     source          1..5054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    186..325
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(251..270)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     411..433
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(427..618)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1127..1146
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1127..1145
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1177..1199
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     sig_peptide     1207..1272
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             2681..2698
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2751..2769)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2765..2812
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2849..3304
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2936..2955)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3146..3167
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3330..3434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3435..3503,3504..4295)
                     /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(3653..3672)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4466..5054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4955..4974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
      241 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
      301 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
      361 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
      421 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
      481 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
      541 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
      601 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
      661 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
      721 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
      781 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
      841 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
      901 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
      961 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     1021 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     1081 ccgtggccag gacccaacgc tgcccgagat ctcgatcccg cgaaattaat acgactcact
     1141 atagggagac cacaacggtt tccctctaga aataattttg tttaacttta agaaggagat
     1201 atacatatga aatacctgct gccgaccgct gctgctggtc tgctgctcct cgctgcccag
     1261 ccggcgatgg ccatggatgt ctagtactca agtaggaaat gctctatcta gttccactac
     1321 tactttagtg gacttgtcta attctacggt tacccaaaag aagcaatatt ataaagatgg
     1381 cgagacgctg cacaatcttt tgcttgaact aaagaataac caagatttgg aacttttacc
     1441 gcatgaacaa gcgcatccta aaatatttca agcgctcaag gctcgtattg gtagaattaa
     1501 taatgaaacg tgcgaccccg gtgaggagaa ctcgtttttc atatgcgatt tgggagaagt
     1561 caagagatta ttcaacaact gggtgaagga gcttcctaga attaagccat tttatgccgt
     1621 caaatgtaat cctgatacca aggttttgtc attattagca gagttgggcg ttaatttcga
     1681 ttgcgcttcc aaagtggaaa ttgacagagt attatcgatg aacatctcgc cggatagaat
     1741 tgtttacgct aatccttgta aagtagcatc tttcattaga tatgcagctt caaaaaatgt
     1801 aatgaagtct acttttgaca atgtagaaga attgcataaa atcaaaaagt ttcatcctga
     1861 gtctcagttg ttattaagaa tcgctaccga tgactctacc gctcaatgtc gactttccac
     1921 caaatatggc tgtgaaatgg aaaacgtaga cgttttatta aaggctataa aggaactagg
     1981 tttaaacctg gctggtgttt ctttccacgt cggttcaggc gcttctgatt ttacaagctt
     2041 atacaaagcc gttagagatg caagaacggt atttgacaaa gctgctaacg aatacgggtt
     2101 gccccctttg aagattttgg atgtaggtgg tggatttcaa tttgaatcct tcaaagaatc
     2161 aactgctgtt ttgcgtctag cgctagagga atttttccct gtaggttgtg gtgttgatat
     2221 aattgcagag cctggcagat actttgtagc tacagcgttc actttggcat ctcatgtgat
     2281 tgcgaagaga aaactgtctg agaatgaagc aatgatttac actaacgatg gtgtatacgg
     2341 gaacatgaat tgtattttat tcgatcatca agagccccat ccaagaaccc tttatcataa
     2401 tttggaattt cattacgacg attttgaatc cactactgcg gtcctcgact ctatcaacaa
     2461 aacaagatct gagtatccat ataaagtttc catctgggga cccacatgtg atggtttgga
     2521 ttgtattgcc aaagagtatt acatgaagca tgatgttata gtcggtgatt ggttttattt
     2581 tcctgccctg ggtgcctaca catcatcggc ggctactcaa ttcaacggct ttgagcagac
     2641 tgcggatata gtatacatag actctgaact cgatctcgag caccaccacc accaccactg
     2701 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     2761 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     2821 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     2881 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2941 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3001 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3061 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3121 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3181 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3241 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     3301 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3361 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3421 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3481 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3541 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3601 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3661 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3721 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3781 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3841 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3901 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3961 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     4021 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4081 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4141 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4201 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4261 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4321 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4381 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4441 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4501 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4561 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4621 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4681 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4741 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4801 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4861 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4921 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4981 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5041 ggagcctatg gaaa
//
