LOCUS       sequence-254604-        7010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SaODH and opine dehydrogenase from Staphylococcus aureus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7010)
  AUTHORS   McFarlane JS, Lamb AL
  TITLE     Biosynthesis of an Opine Metallophore by Pseudomonas aeruginosa.
  JOURNAL   Biochemistry. 2017 Nov 14;56(45):5967-5971. doi: 
            10.1021/acs.biochem.7b00804. Epub 2017 Nov 3.
  PUBMED    29091735
REFERENCE   2  (bases 1 to 7010)
  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: 
            "10.1021/acs.biochem.7b00804"; journalName: "Biochemistry"; date: 
            "2017-11-14- 14"; volume: "56"; issue: "45"; pages: "5967-5971"
COMMENT     Sequence Label: pET15TV-L-SaODH
FEATURES             Location/Qualifiers
     source          1..7010
                     /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(1511..1529)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1525..1572
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1747..1752,1753..1769,1770..1775))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1802..1820)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1888..1992
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1993..2061,2062..2853)
                     /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(2211..2230)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3024..3612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3513..3532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3766..3783
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3798..3937
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3863..3882)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4023..4045
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4039..4230)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5542..6624)
                     /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     6586..6605
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6625..6702)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6908..6927
                     /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 catatgagca agctgctgat gattggcacc
      181 ggtccggtgg cgatccagct ggcgaacatt tgctatctga aaagcgacta cgagattgat
      241 atggttggtc gtgcgagcac cagcgagaag agcaagcgtc tgtaccaggc gtataagaaa
      301 gagaagcaat tcgaagtgaa aatccagaac gaggcgcacc aacacctgga gggcaagttc
      361 gaaatcaacc gtctgtacaa ggacgttaag aacgtgaaag gcgagtacga aaccgtggtt
      421 atggcgtgca ccgcggacgc gtactatgat accctgcagc aactgagcct ggaaaccctg
      481 caaagcgtta agcacgtgat cctgattagc ccgaccttcg gtagccagat gatcgttgag
      541 caattcatga gcaaatttag ccaggacatc gaagtgatta gctttagcac ctatctgggc
      601 gacacccgta ttgttgataa agaggcgccg aaccacgttc tgaccaccgg tgtgaagaaa
      661 aagctgtaca tgggcagcac ccacagcaac agcaccatgt gccaacgtat cagcgcgctg
      721 gcggagcagc tgaaaattca actggaagtg gttgaaagcc cgctgcacgc ggaaacccgt
      781 aacagcagcc tgtacgtgca cccgccgctg ttcatgaacg actttagcct gaaggcgatc
      841 ttcgagggca ccgatgtgcc ggtttacgtg tataaactgt ttccggaagg cccgatcacc
      901 atgaccctga ttcgtgagat gcgtctgatg tggaaggaaa tgatggcgat cctgcaagcg
      961 ttccgtgttc cgagcgtgaa cctgctgcag tttatggtta aagagaacta tccggtgcgt
     1021 ccggagaccc tggacgaagg tgatattgag cacttcgaaa tcctgccgga cattctgcag
     1081 gagtacctgc tgtatgttcg ttacaccgcg atcctgattg acccgttcag ccaaccggat
     1141 gaaaacggcc actatttcga ttttagcgcg gttccgttta agcaggtgta caaaaacgag
     1201 caggacgtgg ttcaaatccc gcgtatgccg agcgaagatt actatcgtac cgcgatgatc
     1261 caacacattg gcaagatgct gggcatcaaa accccgatga ttgaccagtt cctgacccgt
     1321 tatgaagcga gctgccaggc gtacaaagac atgcaccaag atcagcaact gagcagccag
     1381 ttcaacacca acctgtttga gggcgataag gcgctggtta ccaaatttct ggaaatcaac
     1441 cgtaccctga gctaactcga ggatccggct gctaacaaag cccgaaagga agctgagttg
     1501 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1561 aggggttttt tgctgaaagg aggaactata tccggatatc ccgcaagagg cccggcagta
     1621 ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga
     1681 gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac
     1741 taccgcatta aagcttatcg atgataagct gtcaaacatg agaattcttg aagacgaaag
     1801 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     1861 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1921 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1981 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2041 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2101 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2161 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2221 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2281 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2341 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2401 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2461 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2521 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2581 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     2641 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     2701 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     2761 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2821 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     2881 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2941 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3001 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3061 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3121 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3181 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3241 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3301 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3361 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3421 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3481 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3541 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3601 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3661 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3721 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3781 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3841 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     3901 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     3961 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4021 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     4081 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     4141 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     4201 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     4261 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     4321 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     4381 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     4441 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     4501 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     4561 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     4621 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     4681 gatcatgcgc acccgtggcc aggacccaac gctgcccgag atgcgccgcg tgcggctgct
     4741 ggagatggcg gacgcgatgg atatgttctg ccaagggttg gtttgcgcat tcacagttct
     4801 ccgcaagaat tgattggctc caattcttgg agtggtgaat ccgttagcga ggtgccgccg
     4861 gcttccattc aggtcgaggt ggcccggctc catgcaccgc gacgcaacgc ggggaggcag
     4921 acaaggtata gggcggcgcc tacaatccat gccaacccgt tccatgtgct cgccgaggcg
     4981 gcataaatcg ccgtgacgat cagcggtcca atgatcgaag ttaggctggt aagagccgcg
     5041 agcgatcctt gaagctgtcc ctgatggtcg tcatctacct gcctggacag catggcctgc
     5101 aacgcgggca tcccgatgcc gccggaagcg agaagaatca taatggggaa ggccatccag
     5161 cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt cggccgccat gccggcgata
     5221 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
     5281 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
     5341 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
     5401 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
     5461 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     5521 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5581 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     5641 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     5701 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     5761 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     5821 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     5881 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     5941 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     6001 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     6061 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     6121 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     6181 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     6241 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     6301 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     6361 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     6421 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     6481 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     6541 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     6601 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     6661 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
     6721 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     6781 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     6841 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     6901 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     6961 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
//
