LOCUS       sequence-211979-        4614 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 4614)
  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 4614)
  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..4614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..34
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     19..35
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1770..1789)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1771..1789)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1799..1816)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1799..1815)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1808..1830)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1957..2412
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2044..2063)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2254..2275
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2438..2542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2543..2611,2612..3403)
                     /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(2761..2780)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3574..4162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4063..4082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4316..4333
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4450..4471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4486..4491,4492..4509,4510..4516)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4524..4540
                     /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     4529..4551
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4548..4564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4548..4564
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4583..4603
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4585..4603
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gggtaccggg ccccccctcg aggtcgacgg tatcgataag cttgatatcg aattcgatcc
       61 cgcatgcctt aggttttcta gttggatttg ctcccccgcc gtcgttcaat gagaatggat
      121 aagaggctcg tgggattgac gtgagggggc agggatggct atatttctgg gagcgaactc
      181 ccgggcgaat acgaagcgct tggatacagt tgtagggagg gatccatatg actaaggaaa
      241 ataagccaag agatattgtc atcgttggcg ctggcgttat tggattgacc actgcttgga
      301 ttctttcaga cttgggtctt gctcctcgta ttaaggtgat tgccaagtat acgcctgaag
      361 atcgttctgt agaatacact tccccttggg ctggcgcaaa tttctgtagc atttctgcta
      421 ctgatgacaa tgctttgcgc tgggataaaa tcacttacca tcgtttcgcc tacttggcga
      481 aaactcgtcc tgaagcagga atccgttttg ctgatcttcg agaattgtgg gagtacgagc
      541 cgaaacacga caaaatcaga tcctggaata cctatgtcag agatttcaaa gttatccctg
      601 aaaaagatct tccaggagaa tgtatctacg gacataaggc caccaccttt ttaatcaacg
      661 ctcctcatta cttgaattat atgtacaagc tgctcattga agctggcgtc gaatttgaaa
      721 agaaagaatt gagtcacatc aaagagactg tcgaagaaac tccagaagct tcagtagtat
      781 ttaattgcac tggtctctgg gcttccaaat tgggtggcgt tgaagacccg gacgtttatc
      841 cgactcgtgg acatgttgtt ttggttaagg ctcctcatgt aacagaaact cgcattttga
      901 atggcaagaa ctctgatacc tatattattc ctcgtccctt aaatggtgga gtcatttgcg
      961 gcggtttcat gcaaccagga aactgggatc gtgaaattca ccctgaagac actttggata
     1021 tccttaagag aacatcggct ttgatgccag aattgttcca cggcaagggt ccggagggtg
     1081 ctgaaattat tcaagaatgt gtcggattcc gtccttctcg aaagggtggt gcccgcgtag
     1141 agcttgatgt tgttcccggc acctcagtcc cccttgttca tgattacggt gcttctggca
     1201 caggatacca agctggttat ggtatggctc ttgactctgt catgttggct cttcctaaaa
     1261 tcaaattggc ttaactgcag gcatgcaagc ttgtactcaa gctcgtaacg aaggtcgtga
     1321 ccttgctcgt gaaggtggcg acgtaattcg ttcagcttgt aaatggtctc cagaacttgc
     1381 tgctgcatgt gaagtttgga aagaaattaa attcgaattt gatactattg acaaacttta
     1441 atttttattt ttcatgatgt ttatgtgaat agcataaaca tcgtttttat ttttatggtg
     1501 tttaggttaa atacctaaac atcattttac atttttaaaa ttaagttcta aagttatctt
     1561 ttgtttaaat ttgcctgtct ttataaatta cgatgtgcca gaaaaataaa atcttagctt
     1621 tttattatag aatttatctt tatgtattat attttataag ttataataaa agaaatagta
     1681 acatactaaa gcggatgtaa ctcaatcggt agagtgcgat ccactagttc tagctagagc
     1741 ggccgccacc gcggtggagc tccaattcgc cctatagtga gtcgtattac gcgcgctcac
     1801 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     1861 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     1921 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     1981 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2041 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2101 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2161 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2221 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2281 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     2341 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     2401 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2461 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2521 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2581 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2641 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2701 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2761 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2821 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2881 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2941 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3001 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3061 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3121 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3181 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3241 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3301 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3361 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3421 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3481 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3541 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3601 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3661 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3721 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3781 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3841 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3901 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3961 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4021 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4081 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4141 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4201 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4261 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4321 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     4381 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     4441 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     4501 cccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     4561 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agct
//
