LOCUS       sequence-424246-        3931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  integration vector for three gene cassette with integration site E3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3931)
  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 3931)
  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: pCfB12715
FEATURES             Location/Qualifiers
     source          1..3931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(241..1032,1033..1101))
                     /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     864..883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1102..1206)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1309..1897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1798..1817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             2707..2718
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2725..3438
                     /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"
     terminator      3450..3521
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3537..3564
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 gtacagtaca ggcacagtaa ggtgacacgt agtagaacag tcaaactaac acatcaggcc
       61 caccatccac cttacacgta cggtacttgt agaaaagcgc ttcgttcaat tcaggtttta
      121 ggcatttcct tcacgcggcc gctgcggatt atcaaaaagg atcttcacct agatcctttt
      181 aaattaaaaa tgaagtttta aataaatcta aagtatatat gagtaaactt ggtctgacag
      241 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
      301 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      361 cagtgctgca atgataccgc gggacccacg ctcaccggct ccagatttat cagcaataaa
      421 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      481 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      541 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      601 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      661 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      721 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      781 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      841 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      901 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      961 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     1021 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     1081 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     1141 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     1201 tccgcgcaca tttccccgaa aagtgccacc tgtcatgacc aaaatccctt aacgtgagtt
     1261 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1321 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1381 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1441 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1501 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1561 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1621 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1681 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1741 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1801 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     1861 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     1921 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     1981 ttctgtggat aaccgtgcct gcggccgcta acttttttcg tgactctgtt ctccacggtc
     2041 aggtgtcgta tacgtaagta ctgatactcg ctcgtactgg tactgtgcgg tacatacaca
     2101 gtacgttggc accagtgaca attgtgctat cgatactgtt ataagcggct tctatccatg
     2161 gtttactgag gagaaatgtt ctagcagtga cgttacaagt atcgtcacta cattatgttg
     2221 ccatgttcgc acaatacaac atacagaccc gttttcaatg cgacgtatac gttcaggtac
     2281 cttcagtgca tgtgtatgta ccacattata cgaaacagct cgcactatat accatgcaac
     2341 ttgtacaagt atgggccaca ttgtgacacc aatacagaac gcacaaacgg tacaccgacg
     2401 caggaagttt ggaagagcgt tcatgccgat tgtgtactca cagtaccagt acaatacacc
     2461 ggtacttctt ccctgtatcg tacgaacttg tactcctagt ctaaatgacc tcaaactcaa
     2521 gacatcaaac aacggaacgt gagaccactg atgagacgga aagtgaaacg tgatttcatg
     2581 cgtcattttg aacattttgt aaatcttatt taataatgtg tgcggcaatt cacatttaat
     2641 ttatgaatgt tttcttaaca tcgcggcaac tcaagaaacg gcaggttcgg atcttagcta
     2701 ctagagaaag aggagaaata ctagatgcgt aaaggcgaag agctgttcac tggtgtcgtc
     2761 cctattctgg tggaactgga tggtgatgtc aacggtcata agttttccgt gcgtggcgag
     2821 ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt tcatctgtac tactggtaaa
     2881 ctgccggttc cttggccgac tctggtaacg acgctgactt atggtgttca gtgctttgct
     2941 cgttatccgg accatatgaa gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat
     3001 gtgcaggaac gcacgatttc ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg
     3061 aaatttgaag gcgataccct ggtaaaccgc attgagctga aaggcattga ctttaaagag
     3121 gacggcaata tcctgggcca taagctggaa tacaatttta acagccacaa tgtttacatc
     3181 accgccgata aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca caacgtggag
     3241 gatggcagcg tgcagctggc tgatcactac cagcaaaaca ctccaatcgg tgatggtcct
     3301 gttctgctgc cagacaatca ctatctgagc acgcaaagcg ttctgtctaa agatccgaac
     3361 gagaaacgcg atcatatggt tctgctggag ttcgtaaccg cagcgggcat cacgcatggt
     3421 atggatgaac tgtacaaatg accaggcatc aaataaaacg aaaggctcag tcgaaagact
     3481 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     3541 caccttcggg tgggcctttc tgcgtttata cgtctcagtt cggtctcagc tggagttctg
     3601 gcacagaaat gaccacacat acccagttaa ctaagccatt ttcttcatca gcgatggaaa
     3661 tgcatttaaa gatggttcta tttcatacaa ctacccattg actaatggtt attcttgtaa
     3721 aatgatggag aacttctact acaagtacaa atactacacg cgaagttccc cagattgcac
     3781 cgccacaaac accaatcgaa ggtttctctc catacatttt tcccatttgg gacctgtgca
     3841 taattataat tcatcaccat aaaaatttcc tgttgcataa caacctgact cctgaagttt
     3901 cgcgcatgtg tacatattgc tccatcttca a
//
