LOCUS       sequence-240901-        7079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YpODH and opine dehydrogenase from Yersinia pests.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7079)
  AUTHORS   McFarlane JS, Davis CL, Lamb AL
  TITLE     Staphylopine, pseudopaline and yersinopine dehydrogenases: a 
            structural and kinetic analysis of a new functional class of opine 
            dehydrogenase.
  JOURNAL   J Biol Chem. 2018 Apr 4. pii: RA118.002007. doi: 
            10.1074/jbc.RA118.002007.
  PUBMED    29618515
REFERENCE   2  (bases 1 to 7079)
  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"; journalName: "J Biol 
            Chem. 2018 Apr 4. pii: RA118.002007. doi: 10.1074/jbc.RA118.002007."
COMMENT     Sequence Label: pET15TV-L-YpODH
FEATURES             Location/Qualifiers
     source          1..7079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             130..150
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1580..1598)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1594..1641
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1816..1821,1822..1838,1839..1844))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1871..1889)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1957..2061
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2062..2130,2131..2922)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2280..2299)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3093..3681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3582..3601
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3835..3852
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3867..4006
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3932..3951)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4092..4114
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4108..4299)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5611..6693)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6655..6674
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6694..6771)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6977..6996
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcagag aaaacttgta tttccagggc catatgcaca acaccctgcc gaccctgatt
      181 ctgggtgcgg gtccggcggc gatccagctg gcggtggaca ttagcgcgac cggtgatgcg
      241 cgtctgggtc tgtacaaccg tccgagcacc aagggcgagc gtctgaaaca gtacctggcg
      301 ctgaccccga ccctgtatct gcaaggcacc ggcaaggcgc aggcgaccca aaaagaaagc
      361 agcgtgacca tcgactgcta cattgatcag ctggcgcaag cggttggcga ttggcaacgt
      421 ctgatcctgg cggtgccggc ggaccactac tatgcggttc tgcagcaaat tccgtgggcg
      481 gcgctgccgc agctgaagag cgtgatcctg ctgagcagca gcatgggtag cggcctgatg
      541 gttcaaaacc tgctgaacgc ggcgggcaag cgtgatgttg aggtgatcag cctgagcagc
      601 tactatgcgg acaccaaata catccgtgcg gaaacccagg acattagcgc gaacacccaa
      661 gacatcaacg cgggcaccca ggacatcggc gcgattcaac cgtaccgtgc gtataccaag
      721 gcgttcaaac agcgtatcta tctggcgaac caatggggta acgcgggcag cgcggaaatg
      781 agctggctga ccgcggtgct ggcgcgtcac cacattgata ccctgccgtg cagcaacctg
      841 ctggcggcgg aacgtttcag catcaccaac tacgttcacc cgccgctggc gctggcggac
      901 accaccctgc aggcgctgtt ttatccggag cagcgtagcc aatacctgta taagacccag
      961 ccggaaggtc cggtgtgccc ggcggttatt gcggatctgg cgggcctggc ggacgattac
     1021 aaacgtctgc tgaaccgtct gggtgttgag gaaatcaacc tgctgcgttt tctgaacgac
     1081 gataactatc cggtgccggc gagcatggtt agccgtcgtt ggattgacga gttcccgcag
     1141 ctgccgccgc tggaacagca atacgcgctg tttgtgcgtt ataccgcgct gctggttgac
     1201 ccgtacagca ccccggatga gcagggtcgt ttctatgatt ttagcgcggt gaaggttgcg
     1261 accgtgtacc aagacgcgaa cgcgctgtgg cacctgccgc gtgtgccgct ggaagatgtt
     1321 cacaaactgc gtaccctgct gctgctggcg ggtgcgctgg acgtggttat gccgaccgcg
     1381 cagcgtctgc tgcaacgttt ccagcaagcg ctgaaagcgt ttatcgatcg tgttggcgag
     1441 gaacactgcc atccgagcct gctgggtgat gattgcgacc gtcaggcggc gatcattgag
     1501 cagcaatggc gtagccaaac ctaactcgag gatccggctg ctaacaaagc ccgaaaggaa
     1561 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     1621 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc cgcaagaggc
     1681 ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga
     1741 tgacgatgag cgcattgtta gatttcatac acggtgcctg actgcgttag caatttaact
     1801 gtgataaact accgcattaa agcttatcga tgataagctg tcaaacatga gaattcttga
     1861 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     1921 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1981 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2041 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2101 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2161 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2221 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2281 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     2341 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2401 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2461 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2521 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2581 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     2641 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2701 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2761 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2821 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2881 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2941 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3001 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3061 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3121 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3181 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3241 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3301 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3361 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3421 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3481 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3541 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3601 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3661 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3721 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3781 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3841 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     3901 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3961 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     4021 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4081 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4141 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     4201 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     4261 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     4321 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     4381 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     4441 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     4501 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     4561 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     4621 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     4681 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     4741 caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt
     4801 gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt
     4861 cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag
     4921 gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg
     4981 gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc
     5041 gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt taggctggta
     5101 agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc
     5161 atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag
     5221 gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg
     5281 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     5341 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     5401 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     5461 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     5521 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     5581 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5641 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5701 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     5761 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     5821 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     5881 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     5941 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     6001 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     6061 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     6121 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     6181 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     6241 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     6301 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     6361 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     6421 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     6481 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     6541 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     6601 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     6661 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     6721 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     6781 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     6841 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     6901 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     6961 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     7021 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaat
//
