LOCUS       sequence-424068-        4016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  integration vector for single cassette with integration site E3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4016)
  AUTHORS   Jiang W, Wang S, Avila P, Jorgensen TS, Yang Z, Borodina I
  TITLE     Combinatorial iterative method for metabolic engineering of Yarrowia
            lipolytica: application for betanin biosynthesis.
  JOURNAL   Metab Eng. 2024 Sep 9:S1096-7176(24)00110-1. doi: 
            10.1016/j.ymben.2024.09.003.
  PUBMED    39260817
REFERENCE   2  (bases 1 to 4016)
  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: "Metab Eng. 
            2024 Sep 9:S1096-7176(24)00110-1. doi: 10.1016/j.ymben.2024.09.003."
COMMENT     Sequence Label: pCfB13014
FEATURES             Location/Qualifiers
     source          1..4016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             364..1155
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      complement(1696..1723)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1739..1810)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1822..2535)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     RBS             2542..2553
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(3362..3950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3442..3461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttggtcatga ccagccagga cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg
       61 tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc ctgttcggtt
      121 cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa
      181 cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttggg
      241 gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg
      301 atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac
      361 attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc
      421 atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat
      481 ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat
      541 gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag
      601 agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg
      661 cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgcg
      721 ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga
      781 gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa
      841 caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg
      901 gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc
      961 ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag
     1021 tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg
     1081 gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag
     1141 gtagtcggca aataatcaga attggttaat tggttgtaac accgcagcgg ccgcgtgaag
     1201 gaaatgccta aaacctgaat tgaacgaagc gcttttctac aagtaccgta cgtgtaaggt
     1261 ggatggtggg cctgatgtgt tagtttgact gttctactac gtgtcacctt actgtgcctg
     1321 tactgtactt gaagatggag caatatgtac acatgcgcga aacttcagga gtcaggttgt
     1381 tatgcaacag gaaattttta tggtgatgaa ttataattat gcacaggtcc caaatgggaa
     1441 aaatgtatgg agagaaacct tcgattggtg tttgtggcgg tgcaatctgg ggaacttcgc
     1501 gtgtagtatt tgtacttgta gtagaagttc tccatcattt tacaagaata accattagtc
     1561 aatgggtagt tgtatgaaat agaaccatct ttaaatgcat ttccatcgct gatgaagaaa
     1621 atggcttagt taactgggta tgtgtggtca tttctgtgcc agaactccgt ctccttggac
     1681 tctgagacct ataaacgcag aaaggcccac ccgaaggtga gccagtgtga ctctagtaga
     1741 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc
     1801 gttttatttg atgcctggtc atttgtacag ttcatccata ccatgcgtga tgcccgctgc
     1861 ggttacgaac tccagcagaa ccatatgatc gcgtttctcg ttcggatctt tagacagaac
     1921 gctttgcgtg ctcagatagt gattgtctgg cagcagaaca ggaccatcac cgattggagt
     1981 gttttgctgg tagtgatcag ccagctgcac gctgccatcc tccacgttgt ggcgaatttt
     2041 aaaattcgct ttaatgccat ttttttgttt atcggcggtg atgtaaacat tgtggctgtt
     2101 aaaattgtat tccagcttat ggcccaggat attgccgtcc tctttaaagt caatgccttt
     2161 cagctcaatg cggtttacca gggtatcgcc ttcaaatttc acttccgcac gcgttttgta
     2221 cgtgccgtca tccttaaagg aaatcgtgcg ttcctgcaca tagccttccg gcatggcgga
     2281 cttgaagaag tcatgctgct tcatatggtc cggataacga gcaaagcact gaacaccata
     2341 agtcagcgtc gttaccagag tcggccaagg aaccggcagt ttaccagtag tacagatgaa
     2401 cttcagcgtc agtttaccat tagttgcgtc accttcaccc tcgccacgca cggaaaactt
     2461 atgaccgttg acatcaccat ccagttccac cagaataggg acgacaccag tgaacagctc
     2521 ttcgccttta cgcatctagt atttctcctc tttctctagt agctaagatc cgaacctgcc
     2581 gtttcttgag ttgccgcgat gttaagaaaa cattcataaa ttaaatgtga attgccgcac
     2641 acattattaa ataagattta caaaatgttc aaaatgacgc atgaaatcac gtttcacttt
     2701 cggtctcccc gttcagcgag acgccgttgt ttgatgtctt gagtttgagg tcatttagac
     2761 taggagtaca agttcgtacg atacagggaa gaagtaccgg tgtattgtac tggtactgtg
     2821 agtacacaat cggcatgaac gctcttccaa acttcctgcg tcggtgtacc gtttgtgcgt
     2881 tctgtattgg tgtcacaatg tggcccatac ttgtacaagt tgcatggtat atagtgcgag
     2941 ctgtttcgta taatgtggta catacacatg cactgaaggt acctgaacgt atacgtcgca
     3001 ttgaaaacgg gtctgtatgt tgtattgtgc gaacatggca acataatgta gtgacgatac
     3061 ttgtaacgtc actgctagaa catttctcct cagtaaacca tggatagaag ccgcttataa
     3121 cagtatcgat agcacaattg tcactggtgc caacgtactg tgtatgtacc gcacagtacc
     3181 agtacgagcg agtatcagta cttacgtata cgacacctga ccgtggagaa cagagtcacg
     3241 aaaaaagtta gcggccgcag gcacggttat ccacagaatc aggggataac gcaggaaaga
     3301 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3361 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3421 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3481 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3541 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3601 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3661 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3721 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3781 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3841 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3901 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3961 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggatt
//
