LOCUS       sequence-199848-        4177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid vector pDGB2-alpha-1, adapted for S. cerevisiae genome 
            integration by flanking the GB cassette with recombination arms of 
            homology for integration at the YPRC-Delta-15 solo LTR loci.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4177)
  AUTHORS   Perez-Gonzalez A, Kniewel R, Veldhuizen M, Verma HK, 
            Navarro-Rodriguez M, Rubio LM, Caro E
  TITLE     Adaptation of the GoldenBraid modular cloning system and creation of
            a toolkit for the expression of heterologous proteins in yeast 
            mitochondria.
  JOURNAL   BMC Biotechnol. 2017 Nov 13;17(1):80. doi: 
            10.1186/s12896-017-0393-y.
  PUBMED    29132331
REFERENCE   2  (bases 1 to 4177)
  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: "BMC 
            Biotechnol."; date: "2017-11-13"; volume: "17(1):80. doi"; pages: " 
            10.1186/s12896-017-0393-y"
COMMENT     Sequence Label: YPRC-Delta-15-alpha-1
FEATURES             Location/Qualifiers
     source          1..4177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1066..1389)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1071..1090
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1284..1306
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1298..1315
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1299..1315
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1316..1372
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1385..1401)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1385..1401)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1398..1420)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1409..1425
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1433..1439,1440..1457,1458..1463))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1478..1499
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2616..3204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2696..2715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3292..4107)
                     /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(3292..4107)
                     /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     4015..4034
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gacgaactcc aattcgcggc cgctagggat aacagggtaa tcgagatagt ttgacgttcg
       61 ttttttactt tgaatatact cgtagtcttt ttactttttg agtataaaaa aaaaatgact
      121 agcaaaataa aattagtagt ctaaaaaaga aagctcgcac tcaggatcga actaaggacc
      181 aacagatttg caatctgctg cgctaccact gcgccatacg agcttttgaa ttatggtaat
      241 tttgattatc ctagaatgtt atatctcaat atctcaatat attttggaca tctatgaaac
      301 acccataaag cagccgctac caaacagaca agattcagta tgtaaggtaa ataccttttt
      361 gcacagttaa actacccaaa cttattaaag cttgataaat tactgaaatt ccacctttca
      421 gttagattca ggcctcatat agattagata tagggtacgt aacattctgt caaccaagtt
      481 gttggaatga aagtctaaaa tgtcatctat tcggtagcac tcatgttact agtatactgt
      541 cacatgcggt gtaacgtggg gacataaaac agacatcaaa tataatggaa gctgaaatgc
      601 aaagatcgat aatgtaatag gaatgaaaca tataaaacga aaggagaagt aatggtaata
      661 ttagtatgta gaaataccga ttcaattttg gggattctta tattctcgag agaatttcta
      721 gtataatctg tatacataat attataggct ttaccaacaa tggaatttcg acaattatca
      781 tattattcac caattaatca caagttggta atgagtttga taacaagtta ctttcttaac
      841 aacgttagta tcgtcaaaac actcggtttt actcgagctt gtagcacaat aataccgtgt
      901 agagttctgt attgttcttc ttagtgcttg tatatgctca tcccgacctt ccattgaatt
      961 cgtctcagga gagagaccca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1021 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     1081 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     1141 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     1201 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     1261 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     1321 cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc ttggcgtaat
     1381 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1441 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1501 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctggggtctc
     1561 tcgctgtcat gagacgaatt ctttgcgaaa ccctatgctc tgttgttcgg atttgaaatt
     1621 ttaaaactac attaatgtgt tagtttttct ttctttcttt ctttgtcttg acgtgatttg
     1681 gacttctgtc ttgcattcgc gtccattcat ctgacccaat attccttttg gttttgttat
     1741 ccttataaaa agaaaggaag cttcttagag ggaaaaaaat gatgaagagt aatgccaaaa
     1801 tataaataaa taaataaata tgaaaatcat tttctatttt taatagaata agaagagcat
     1861 cttaagatta caatttcaag aaatagttta cacagtatat ccaataactc caataaacta
     1921 ctttcctata caaatttcta tggtgggatt aatagtaaaa cttctgtact tctctaattc
     1981 accaagaaat taaggtaaac atctggtaag cactatccag ctttttgcta ttacacatat
     2041 ggcttttctg caatcatttc ttcccatttt gtctcaagcc gttagtcttg aaaccacagg
     2101 cggagtagag ttacttgatg cggtatttta catgcctttt ttcactgcaa aaaaaatgaa
     2161 atacatattt acacgatttg caggacagtt tacgatagtg agtatgcaga atagttaaca
     2221 cctttgtttt atccttttgt gtcttaatta tatgatataa aggcgcctgg cgttatcgga
     2281 tagtaataga tgctagttat caacatttca caattgaagg aaataaagtt gaagtactca
     2341 acaaaaactt acttcagaat taaatttttg gggggaacat aggcatccta tgacaggtga
     2401 ccacaagccc ctcaacgcaa tctaatattt tacaaagtgg taaaattctt tcgttcttcg
     2461 ttttaatata cagtcattta ttgattctat tacattaata ttcctacgct tcggctcaca
     2521 taattaacag gacttcgagt ccgttaaact tgggatcaac taatttctac ggattaggga
     2581 taacagggta atgcggccgc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2641 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2701 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2761 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2821 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2881 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2941 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3001 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     3061 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3241 cagtggaacg aaaactcacg ttaagggatt ttggtcatgc attctaggtg attagaaaaa
     3301 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     3361 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     3421 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     3481 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     3541 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     3601 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     3661 gagtcgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     3721 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     3781 tacctggaat gctgttttcc ctgggatcgc agtggtgagt aaccatgcat catcaggagt
     3841 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     3901 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     3961 cgcatcgggc ttcccataca atcggtagat tgtcgcacct gattgcccga cattatcgcg
     4021 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gccttgagca
     4081 agacgtttcc cgttgaatat ggctcataac agaacttatt atttccttcc tcttttctac
     4141 agtatttaaa gataccccaa gaagctaatt ataacaa
//
