LOCUS       sequence-248557-        7839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid containing integration cassette at IntD1 integration site in
            Yarrowia lipolytica for overexpression of XdCrtI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7839)
  AUTHORS   Milne N, Tramontin LRR, Borodina I
  TITLE     A teaching protocol demonstrating the use of EasyClone and 
            CRISPR/Cas9 for metabolic engineering of Saccharomyces cerevisiae 
            and Yarrowia lipolytica.
  JOURNAL   FEMS Yeast Res. 2019 Sep 26. pii: foz062. doi: 
            10.1093/femsyr/foz062.
  PUBMED    31556952
REFERENCE   2  (bases 1 to 7839)
  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"; journalName: "FEMS Yeast 
            Res. 2019 Sep 26. pii: foz062. doi: 10.1093/femsyr/foz062."
COMMENT     Sequence Label: pCfB8786
FEATURES             Location/Qualifiers
     source          1..7839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..34
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(5096..5129)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(5599..5616)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5599..5615)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5714..5731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5885..6473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5965..5984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6595..7386,7387..7455))
                     /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     7218..7237
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7456..7547)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7589..7608
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7614..7633
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gaagttccta ttctctagaa agtataggaa cttctgaagt ggggatttaa atgcggccgc
       61 gaatgcacgc gatccggcca ggggtaagag tggagaggtg tggagggtgc aatgggccac
      121 cgagtcggtg tggttgccag aaaacacgcc tcgagcttct tgagcgttat ataatggccg
      181 ccgtttccac aaggctcagc aactcagccg tcgcgtcgtc gcccgtcaac ctcgctttca
      241 tgttggctat aatatcgctt ggttggaaac actaattatg ccatcgcatt gacactcgta
      301 aacctgacac tattgcgcga gggtttgggc atgtcggaga gcagctgcgt ggatacaagg
      361 tgcgacacgg ccacatggca catgtgctag aatgaaccgt tgcaatggcg accacacgcg
      421 ccggggtgtc ggcgttgccc ctgcaaaggt tccggtaacc ggcgtcatcg ctcaaagtcg
      481 atgactgggc cggggagtag atgggcaatg gtgactcagg cggaccgtgg cgaacgagaa
      541 agacacaaac agcagacgca gaaggccact cagatgcatt cttgggcggt ctataccagc
      601 tcattgaccc ttcgtgggtc tcaatgttca acgcagatga acttcaaatg cttatcagtg
      661 gaggacatgc caatgtggac gtctgggatt tgaagacaca caccaattac ggaggctatc
      721 tggatacaga tcaaactata aaggacttct ggactgtgtt tgagagtttc gaggaagagg
      781 acaagcggct gcttcttaag tttgtgacat cagtatccaa ggcaccattg caaggattca
      841 aggctttgaa cccgtcattt gccattcgta acgctggtag acaggttgat cggttcccta
      901 cggcctccac ctgtgtcaat cttctcaagc tgcctgacta tcaggacatt gatcaacttc
      961 ggaagaaact tttgtatgcc attcgatcgc atgctggttt cgatttgtct tagaggaacg
     1021 catatacagt aatcatagag aataaacgat attcatttat taaagtagat agttgaggta
     1081 gaagttgtaa agagtgataa atagcttaga taccacagac accctcggtg acgaagtact
     1141 gcagatggtt tccaatcaca ttgacctgct ggagcagagt gttaccggca gagcactgtt
     1201 tattgctctg gccctggcac atgacaacgt tggagagagg agggtggatc aggggccagt
     1261 caataaagac ctcaccagag cagtgctggt aaccgtccca gaagggcact tgagggacga
     1321 tatctcctcg gtgggtgatt cggtagagct ttcggtcttt ggacaccttg gagacatcgg
     1381 ggttctcctg gccaaagaag agtttatcga cccagttagc aaagccagcg ttaccgacaa
     1441 tgggctgacc aagagtaaca acgaggggat cgtggccgtt aaccttgagg ttgattccga
     1501 acagaaggaa tgcacgcgat gacgcagtag gatgtcctgc acgggtcttt ttgtggggtg
     1561 tggagaaagg ggtgcttgga gatggaagcc ggtagaaccg ggctgcttgg ggggatttgg
     1621 ggccgctggg ctccaaagag gggtaggcat ttcgttgggg ttacgtaatt gcggcatttg
     1681 ggtcctgcgc gcatgtccca ttggtcagaa ttagtccgga taggagactt atcagccaat
     1741 cacagcgccg gatccacctg taggttgggt tgggtgggag cacccctcca cagagtagag
     1801 tcaaacagca gcagcaacat gatagttggg ggtgtgcgtg ttaaaggaaa aaaaaagaag
     1861 cttgggttat attcccgctc tatttagagg ttgcgggata gacgccgacg gagggcaatg
     1921 gcgccatgga accttgcgga tatcgatacg ccgcggcgga ctgcgtccga accagctcca
     1981 gcagcgtttt ttccgggcca ttgagccgac tgcgaccccg ccaacgtgtc ttggcccacg
     2041 cactcatgtc atgttggtgt tgggaggcca ctttttaagt agcacaaggc acctagctcg
     2101 cagcaaggtg tccgaaccaa agaagcggct gcagtggtgc aaacggggcg gaaacggcgg
     2161 gaaaaagcca cgggggcacg aattgaggca cgccctcgaa tttgagacga gtcacggccc
     2221 cattcgcccg cgcaatggct cgccaacgcc cggtcttttg caccacatca ggttacccca
     2281 agccaaacct ttgtgttaaa aagcttaaca tattataccg aacgtaggtt tgggcgggct
     2341 tgctccgtct gtccaaggca acatttatat aagggtctgc atcgccggct caattgaatc
     2401 ttttttcttc ttctcttctc tatattcatt cttgaattaa acacacatca acagtgcagg
     2461 tatgggaaaa gaacaagatc aggataaacc cacagctatc atcgtgggat gtggtatcgg
     2521 tggaatcgcc actgccgctc gtcttgctaa agaaggtttc caggtcacgg tgttcgagaa
     2581 gaacgactac tccggaggtc gatgctcttt aatcgagcga gatggttatc gattcgatca
     2641 ggggcccagt ttgctgctct tgccagatct cttcaagcag acattcgaag atttggggga
     2701 gaagatggaa gattgggtcg atctcatcaa gtgtgaaccc aactatgttt gccacttcca
     2761 cgatgaagag actttcactt tttcaaccga catggcgttg ctcaagcggg aagtcgagcg
     2821 ttttgaaggc aaagatggat ttgatcggtt cttgtcgttt atccaagaag cccacagaca
     2881 ttacgagctt gctgtcgttc acgtcctgca gaagaacttc cctggcttcg cagcattctt
     2941 acggctacag ttcattggcc aaatcctggc tcttcacccc ttcgagtcta tctggacaag
     3001 agtttgtcga tatttcaaga ccgacagatt acgaagagtc ttctcgtttg cagtgatgta
     3061 catgggtcaa agcccataca gtgcgcccgg aacatattcc ttgctccaat acaccgaatt
     3121 gaccgagggc atctggtatc cgagaggagg cttttggcag gttcctaata ctcttcttca
     3181 gatcgtcaag cgcaacaatc cctcagccaa gttcaatttc aacgctccag tttcccaggt
     3241 tcttctctct cctgccaagg accgagcgac tggtgttcga cttgaatccg gcgaggaaca
     3301 tcacgccgat gttgtgattg tcaatgctga cctcgtttac gcctccgagc acttgattcc
     3361 tgacgatgcc agaaacaaga ttggccaact gggtgaagtc aagagaagtt ggtgggctga
     3421 cttagttggt ggaaagaagc tcaagggaag ttgcagtagt ttgagcttct actggagcat
     3481 ggaccgaatc gtggacggtc tgggcggaca caatatcttc ttggccgagg acttcaaggg
     3541 atcattcgac acaatcttcg aggagttggg tctcccagcc gatccttcct tttacgtgaa
     3601 cgttccctcg cgaatcgatc cttctgccgc tcccgaaggc aaagatgcta tcgtcattct
     3661 tgtgccgtgt ggccatatcg acgcttcgaa ccctcaagat tacaacaagc ttgttgctcg
     3721 ggcaaggaag tttgtgatcc aaacgctttc cgccaagctt ggacttcccg actttgaaaa
     3781 aatgattgtg gcagagaagg ttcacgatgc tccctcttgg gagaaagaat ttaacctcaa
     3841 ggacggaagc atcttgggac tggctcacaa ctttatgcaa gttcttggtt tcaggccgag
     3901 caccagacat cccaagtatg acaagttgtt ctttgtcggg gcttcgactc atcccggaac
     3961 tggggttccc atcgtcttgg ctggagccaa gttaactgcc aaccaagttc tcgaatcctt
     4021 tgaccgatcc ccagctccag atcccaatat gtcactctcc gtaccatatg gaaaacctct
     4081 caaatcaaat ggaacgggta tcgattctca ggtccagctg aagttcatgg atttggagag
     4141 atgggtatac cttttggtgt tgttgattgg ggccgtgatc gctcgatccg ttggtgttct
     4201 tgctttctga atcgcacgca ttccgttgaa gtgtggatgg ggaagtgagt gcccggttct
     4261 gtgtgcacaa ttggcaatcc aagatggatg gattcaacac agggatatag cgagctacgt
     4321 ggtggtgcga ggatatagca acggatattt atgtttgaca cttgagaatg tacgatacaa
     4381 gcattgtcca agtacaatac taaacatact gtacatactc atactcgtac ccgggcaacg
     4441 gtttcacttg agtgcagtgg ctagtgccct tactcgtaca gtgtgcaata ctgcgtatca
     4501 tagtctttga tgtatatcgt attcattcat gttagttgcg taagcgttgc acgtgacatt
     4561 gctctaacac tctgcattgt tcccactaat ttgtcaacac tcaacgacca tacccactaa
     4621 ttatgttggt gtagttagac tcgaggtaga tccatggtgc cattctgaaa aaataaataa
     4681 ataaaaacaa ctgatttttc cgctctgata ttttatgtca aaatgattgc gttcatcttc
     4741 cgacttgttc tgttctctct gatcgctcct cgatgtaatc attgtactat acctcacacg
     4801 ccgttatctc ggtattaaca gcaaatgtgc aagccactcg ccccctgaaa cagccgatga
     4861 ggtttcataa gtaattgcgg ccttgtcgag acgctaacag acacatgccc tatgtacccg
     4921 tgctttggag aagttttgcg tcatgcgcgc tagtgagaca tgttgttgag gtgctcgaat
     4981 ccgtgtttgg aatacgttag cgaaccgatg ctaattacga gcgttggtga tgtgtttgat
     5041 atgacgggaa aatgatcgca cgcattcgcg gccgcaaatt taaataaaat gaagtgaagt
     5101 tcctatactt tctagagaat aggaacttct atagtgagtc gaataagggc gacacaaaat
     5161 ttattctaaa tgcataataa atactgataa catcttatag tttgtattat attttgtatt
     5221 atcgttgaca tgtataattt tgatatcaaa aactgatttt ccctttatta ttttcgagat
     5281 ttattttctt aattctcttt aacaaactag aaatattgta tatacaaaaa atcataaata
     5341 atagatgaat agtttaatta taggtgttca tcaatcgaaa aagcaacgta tcttatttaa
     5401 agtgcgttgc ttttttctca tttataaggt taaataattc tcatatatca agcaaagtga
     5461 caggcgccct taaatattct gacaaatgct ctttccctaa actcccccca taaaaaaacc
     5521 cgccgaagcg ggtttttacg ttatttgcgg attaacgatt actcgttatc agaaccgccc
     5581 agggggcccg agcttaagac tggccgtcgt tttacaacac agaaagagtt tgtagaaacg
     5641 caaaaaggcc atccgtcagg ggccttctgc ttagtttgat gcctggcagt tccctactct
     5701 cgccttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5761 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5821 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5881 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5941 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6001 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6061 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6121 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6181 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6241 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6301 gggctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6361 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6421 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6481 ctttgatctt ttctacgggg tctgacgctc agtggaacga cgcgcgcgta actcacgtta
     6541 agggattttg gtcatgagct tgcgccgtcc cgtcaagtca gcgtaatgct ctgcttacca
     6601 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6661 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagcgc
     6721 tgcgatgata ccgcgagaac cacgctcacc ggctccggat ttatcagcaa taaaccagcc
     6781 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6841 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6901 tgccatcgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6961 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7021 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7081 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7141 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7201 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7261 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7321 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7381 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7441 atgttgaata ctcatattct tcctttttca atattattga agcatttatc agggttattg
     7501 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggtcagtgt
     7561 tacaaccaat taaccaattc tgaacattat cgcgagccca tttatacctg aatatggctc
     7621 ataacacccc ttgtttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     7681 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gactccccat gcgagagtag
     7741 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgcccg
     7801 ggctaattat ggggtgtcgc ccttattcga ctctatagt
//
