LOCUS       sequence-170244-        4611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plastid expression cassette containing Schizosaccharomyces pombe 
            D-amino acid oxidase gene. Provides dual positive (D-alanine) and 
            negative (D-valine) selection in transplastomic plants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4611)
  AUTHORS   Gisby MF, Mudd EA, Day A
  TITLE     Growth of transplastomic cells expressing D-amino acid oxidase in 
            chloroplasts is tolerant to D-alanine and inhibited by D-valine.
  JOURNAL   Plant Physiol. 2012 Dec;160(4):2219-26. doi: 10.1104/pp.112.204107. 
            Epub 2012 Oct 18.
  PUBMED    23085840
REFERENCE   2  (bases 1 to 4611)
  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: 
            "10.1104/pp.112.204107"; journalName: "Plant Physiol"; date: 
            "2012-12"; volume: "160"; issue: "4"; pages: "2219-26"
COMMENT     Sequence Label: pUM31dao
FEATURES             Location/Qualifiers
     source          1..4611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..27)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9..27)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(34..51)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(34..50)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(43..65)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      192..647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(279..298)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     489..510
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        673..777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(778..846,847..1638)
                     /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     complement(996..1015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1809..2397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2298..2317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2551..2568
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2685..2706
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2721..2726,2727..2744,2745..2751)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2759..2775
                     /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)."
     primer_bind     2764..2786
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2783..2799
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2783..2799
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2820..2838
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2867..2883
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2868..2884
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga
       61 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
      121 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
      181 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
      241 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      301 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      361 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      421 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      481 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      541 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      601 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
      661 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      721 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      781 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      841 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      901 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      961 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1021 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1081 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     1141 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1201 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1261 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1321 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1381 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1441 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1501 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1561 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1621 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1681 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1741 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1801 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1861 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1921 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     1981 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2041 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2101 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2161 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2221 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2281 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2341 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2401 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2461 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2521 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2581 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     2641 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     2701 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     2761 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctcgaa
     2821 attaaccctc actaaaggga acaaaagctg ggtaccgggc cccccctcga ggtcgacggt
     2881 atcgataagc ttgatatcga attcgatccc gcatgcctta ggttttctag ttggatttgc
     2941 tcccccgccg tcgttcaatg agaatggata agaggctcgt gggattgacg tgagggggca
     3001 gggatggcta tatttctggg agcgaactcc cgggcgaata cgaagcgctt ggatacagtt
     3061 gtagggaggg atccatatga ctaaggaaaa taagccaaga gatattgtca tcgttggcgc
     3121 tggcgttatt ggattgacca ctgcttggat tctttcagac ttgggtcttg ctcctcgtat
     3181 taaggtgatt gccaagtata cgcctgaaga tcgttctgta gaatacactt ccccttgggc
     3241 tggcgcaaat ttctgtagca tttctgctac tgatgacaat gctttgcgct gggataaaat
     3301 cacttaccat cgtttcgcct acttggcgaa aactcgtcct gaagcaggaa tccgttttgc
     3361 tgatcttcga gaattgtggg agtacgagcc gaaacacgac aaaatcagat cctggaatac
     3421 ctatgtcaga gatttcaaag ttatccctga aaaagatctt ccaggagaat gtatctacgg
     3481 acataaggcc accacctttt taatcaacgc tcctcattac ttgaattata tgtacaagct
     3541 gctcattgaa gctggcgtcg aatttgaaaa gaaagaattg agtcacatca aagagactgt
     3601 cgaagaaact ccagaagctt cagtagtatt taattgcact ggtctctggg cttccaaatt
     3661 gggtggcgtt gaagacccgg acgtttatcc gactcgtgga catgttgttt tggttaaggc
     3721 tcctcatgta acagaaactc gcattttgaa tggcaagaac tctgatacct atattattcc
     3781 tcgtccctta aatggtggag tcatttgcgg cggtttcatg caaccaggaa actgggatcg
     3841 tgaaattcac cctgaagaca ctttggatat ccttaagaga acatcggctt tgatgccaga
     3901 attgttccac ggcaagggtc cggagggtgc tgaaattatt caagaatgtg tcggattccg
     3961 tccttctcga aagggtggtg cccgcgtaga gcttgatgtt gttcccggca cctcagtccc
     4021 ccttgttcat gattacggtg cttctggcac aggataccaa gctggttatg gtatggctct
     4081 tgactctgtc atgttggctc ttcctaaaat caaattggct taactgcagg catgcaagct
     4141 tgtactcaag ctcgtaacga aggtcgtgac cttgctcgtg aaggtggcga cgtaattcgt
     4201 tcagcttgta aatggtctcc agaacttgct gctgcatgtg aagtttggaa agaaattaaa
     4261 ttcgaatttg atactattga caaactttaa tttttatttt tcatgatgtt tatgtgaata
     4321 gcataaacat cgtttttatt ttttatggtg tttaggttaa atacctaaac atcattttac
     4381 atttttaaaa ttaagttcta aagttatctt ttgtttaaat ttgcctgtgc tttataaatt
     4441 acgatgtgcc agaaaaataa aatcttagct ttttattata gaatttatct ttatgtatta
     4501 tattttataa gtaataaaag aaatagtaac atactaaagc ggatgtaact caatcggtag
     4561 agtgcgatcc actagtttct agctagagcg gccgccaccg cggtggagct c
//
