LOCUS       sequence-477420-        2796 bp    DNA     circular SYN 30-JUN-2025
DEFINITION  Empty vector for L0 TypeIIS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2796)
  AUTHORS   Tobin EE, Collins JH, Marsan CB, Nadeau GT, Mori K, Lipzen A, Mondo 
            S, Grigoriev IV, Young EM
  TITLE     Omics-driven onboarding of the carotenoid producing red yeast 
            Xanthophyllomyces dendrorhous CBS 6938.
  JOURNAL   Appl Microbiol Biotechnol. 2024 Dec 28;108(1):547. doi: 
            10.1007/s00253-024-13379-w.
  PUBMED    39731599
REFERENCE   2  (bases 1 to 2796)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl 
            Microbiol Biotechnol."; date: "2024-12-28"; volume: "108(1):547. 
            doi"; pages: " 10.1007/s00253-024-13379-w"
COMMENT     Sequence Label: p0AmpCD
FEATURES             Location/Qualifiers
     source          1..2796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..61
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(99..121)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     221..240
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             592..897
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     776..795
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     rep_origin      complement(1047..1635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1127..1146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1806..2597,2598..2666))
                     /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     2429..2448
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2667..2771)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
       61 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
      121 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
      181 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
      241 ctgggtctca taaaaggtct tcggcttact aaaagccaga taacagtatg catatttgcg
      301 cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta tgtcaaaaag
      361 aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta tcagttgctc
      421 aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag aatgaagccc
      481 gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct gaggtcgccc
      541 ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga aatgcagttt
      601 aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat
      661 attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca
      721 gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg
      781 atgaccaccg atatggccag tgtgccggtt tccgttatcg gggaagaagt ggctgatctc
      841 agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataagaa
      901 gacctcctca gagaccgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
      961 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1021 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1081 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1141 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1201 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1261 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1321 cccgaccgct gcgccttatc cggtaactat cgtcttgagc ccaacccggt aagacacgac
     1381 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1441 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1501 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1561 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1621 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1681 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1741 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     1801 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     1861 tccatagttg cctggctccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     1921 ggccccagtg ctgcaatgat accgcgagag ccacgctcac cggctccaga tttatcagca
     1981 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2041 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2101 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2161 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2221 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2281 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2341 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2401 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     2461 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     2521 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     2581 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     2641 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     2701 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2761 ggggttccgc gcacatttcc ccgaaaagtg ccacct
//
