LOCUS       sequence-489330-        4557 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Empty vector for L2 TypeIIS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4557)
  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 4557)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 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: p2KanEG
FEATURES             Location/Qualifiers
     source          1..4557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(284..1099)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(284..1099)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1007..1026
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1091..1110
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2552..2857
                     /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     2736..2755
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     rep_origin      complement(join(4011..4557,1..42))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4091..4110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
       61 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      121 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      181 taaagtatat atgagtaaac ttggtctgac agggggtctg acgctcagtg gaacgaaaaa
      241 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accttagaaa aactcatcga
      301 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
      361 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
      421 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
      481 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
      541 gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
      601 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
      661 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
      721 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
      781 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
      841 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
      901 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
      961 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     1021 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctggag caagacgttt
     1081 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     1141 ttgttcatga tgatatattt ttatcttgtg caatgtacat cagagatttt gagacacaag
     1201 gtctcagctt cagaagatgg gtccaccgat agtggtttga aaggagatgg gtggaataga
     1261 gacaaatggg gaattccagt tttgcctttg acgagaaagg acactgggtt ggaaagagaa
     1321 gatggtacgt tcttctccac cttgaatgtg ttgcttacta gacatgtttg acacgctaat
     1381 gcatttcttt ccactttgac ttttgaacta tggtggttgg gcgatcccca aaatcattag
     1441 cttctacttc agctcattac ctcgatctca tcttactacc aggtgttgca ttctcaccta
     1501 cggcctcttc tttgttctct cgactgggcc atggaaaagg atattacgat aaatacatca
     1561 ctcagtatcg gtcgatctgt gcaggcaaga atcgacccgg tccgaagctg agtacgcgtc
     1621 ttctcttttc tcgataccca acggacgcta ttttgtgaca gaaggatgag actatccaac
     1681 agctcaaaca aactaacgct cttgattaat cacccgctca acttattgct caactcagtt
     1741 ggactggcgc tgaaagaaca gttcttagac aaaaacatgg tccctatagg agaatgggat
     1801 gcgaatctgg atgaagtgtt ggttggagat cacgtgagga cattatccga ggacaattaa
     1861 ctacttaaga tatatacatg atttatgtcg atcggcatcc agccggggat tgatcggctg
     1921 atggccggaa atgtgatgat ggtcgaaact cgatctctct ttttttgttc atcttctcat
     1981 ccctcttctc tctttctact gacatccatc tccaactgtc tagatcagtt cggaaacaag
     2041 aagtggacac agagagatct ttgctgaaga gttgtattcc agaaagggaa aacaaaggaa
     2101 agaagcgccg aagcacatca ccaacttcag caagccggtc cagcccgatc tcggatagac
     2161 atcatcttac ccaactcgta tcatccccaa cagatagagt ttttgtcgca gcttaggtct
     2221 tcggcttact aaaagccaga taacagtatg catatttgcg cgctgatttt tgcggtataa
     2281 gaatatatac tgatatgtat acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg
     2341 tattacagtg acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat
     2401 atctccggtc tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc
     2461 tggaaagcgg aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc
     2521 tcttttgctg acgagaacag gggctggtga aatgcagttt aaggtttaca cctataaaag
     2581 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg
     2641 acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact
     2701 ttacccggtg gtacatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
     2761 tgtgccggtt tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
     2821 caaaaacgcc attaacctga tgttctgggg aatataagaa gaccttacgg acagcggaag
     2881 aataccgaca gacaatgatg agtgagaata aaatcatcct caatcttctt tctctaggtg
     2941 ctcttttttg ttttctatta tgaccaactc taaaggaact ggccttgcag atatttctct
     3001 tccccccatc ttcctccttt ccatcgtttg ttctttccat ttttgtcggt ttactatgtc
     3061 aattcttttt cttgcttttt cttatcaatc tagacaattc tatagatgtt tagaatttat
     3121 acattgacag gttatagacc ataaagacta tatggctttg gctactagag agcaaccgca
     3181 accgatggtg gtcgatacgc ggcccttttg gacttgggtg ccctcagacc acagaccacc
     3241 aagatccccg tgggtcacgg gcttagcttg tgccagcccc agcagtcatg acagtcccag
     3301 ccccagcccc agcctcacta gcaacagcgt caaagcccgg cgtccttgcc aactggttct
     3361 cctcctgatg cagggctagg tgcagcatgt tgcgcgatac gtcgacatac ctgttccacc
     3421 aagtctgaga gaacacagta agcaaacata gagagtagtg agcaaccgac acatcatctt
     3481 gagccataca acacatgtca cttactcgac agacgcttcg acactcctag atcgcccacg
     3541 ctttgcctct tccagagaac tcgccatgac acttcgagca cactctcgac gtccgatact
     3601 cccggacttt gcgagcctgg atcagtaggt agatcctcag cttggcaaac agcaccttga
     3661 caggaccagg agcctgccca cgagaaggcg ccgtcgccga aggccaggaa ggtgcgtgga
     3721 cgtcctgctg acagacggac ggccaccttg cggtacgttt gctcgcggct gatgtgcgcg
     3781 ccccagtgga gacgtctcca ccgccgagcg gcgaagaatg ccgcgagcca agagaaggac
     3841 tgatagcgcg agacgagagc agcatggagt acgagagacc gagcggtatc agctcactca
     3901 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3961 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4021 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4081 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4141 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4201 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4261 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4321 gagcccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4381 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4441 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4501 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg ttttttt
//
