LOCUS       sequence-199843-        4421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid vector pDGB2-Omega-2, 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 4421)
  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 4421)
  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-Omega-2
FEATURES             Location/Qualifiers
     source          1..4421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1237..1560)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1242..1261
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1455..1477
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1469..1486
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1470..1486
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1487..1543
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1556..1572)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1556..1572)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1569..1591)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1580..1596
                     /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(1604..1610,1611..1628,1629..1634))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1649..1670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2783..3371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2863..2882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3459..4250)
                     /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"
ORIGIN
        1 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
       61 ttacagctta cgaaccgaac aggcttatgt ccactgggtt cgtgccttca tccgtttcca
      121 cggtgtgcgt cacccggcaa ccttgggtag cagcgaagtc gaggcatttc tgtcctggtg
      181 ggcggccgct agggataaca gggtaatcga gatagtttga cgttcgtttt ttactttgaa
      241 tatactcgta gtctttttac tttttgagta taaaaaaaaa atgactagca aaataaaatt
      301 agtagtctaa aaaagaaagc tcgcactcag gatcgaacta aggaccaaca gatttgcaat
      361 ctgctgcgct accactgcgc catacgagct tttgaattat ggtaattttg attatcctag
      421 aatgttatat ctcaatatct caatatattt tggacatcta tgaaacaccc ataaagcagc
      481 cgctaccaaa cagacaagat tcagtatgta aggtaaatac ctttttgcac agttaaacta
      541 cccaaactta ttaaagcttg ataaattact gaaattccac ctttcagtta gattcaggcc
      601 tcatatagat tagatatagg gtacgtaaca ttctgtcaac caagttgttg gaatgaaagt
      661 ctaaaatgtc atctattcgg tagcactcat gttactagta tactgtcaca tgcggtgtaa
      721 cgtggggaca taaaacagac atcaaatata atggaagctg aaatgcaaag atcgataatg
      781 taataggaat gaaacatata aaacgaaagg agaagtaatg gtaatattag tatgtagaaa
      841 taccgattca attttgggga ttcttatatt ctcgagagaa tttctagtat aatctgtata
      901 cataatatta taggctttac caacaatgga atttcgacaa ttatcatatt attcaccaat
      961 taatcacaag ttggtaatga gtttgataac aagttacttt cttaacaacg ttagtatcgt
     1021 caaaacactc ggttttactc gagcttgtag cacaataata ccgtgtagag ttctgtattg
     1081 ttcttcttag tgcttgtata tgctcatccc gaccttccat tgatatcggt ctcagtcagg
     1141 agtgagacgc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     1201 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     1261 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     1321 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     1381 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     1441 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcgagctcgg
     1501 tacccgggga tcctctagag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat
     1561 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     1621 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     1681 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcgtctc acgcttgaga
     1741 ccgatatctt tgcgaaaccc tatgctctgt tgttcggatt tgaaatttta aaactacatt
     1801 aatgtgttag tttttctttc tttctttctt tgtcttgacg tgatttggac ttctgtcttg
     1861 cattcgcgtc cattcatctg acccaatatt ccttttggtt ttgttatcct tataaaaaga
     1921 aaggaagctt cttagaggga aaaaaatgat gaagagtaat gccaaaatat aaataaataa
     1981 ataaatatga aaatcatttt ctatttttaa tagaataaga agagcatctt aagattacaa
     2041 tttcaagaaa tagtttacac agtatatcca ataactccaa taaactactt tcctatacaa
     2101 atttctatgg tgggattaat agtaaaactt ctgtacttct ctaattcacc aagaaattaa
     2161 ggtaaacatc tggtaagcac tatccagctt tttgctatta cacatatggc ttttctgcaa
     2221 tcatttcttc ccattttgtc tcaagccgtt agtcttgaaa ccacaggcgg agtagagtta
     2281 cttgatgcgg tattttacat gccttttttc actgcaaaaa aaatgaaata catatttaca
     2341 cgatttgcag gacagtttac gatagtgagt atgcagaata gttaacacct ttgttttatc
     2401 cttttgtgtc ttaattatat gatataaagg cgcctggcgt tatcggatag taatagatgc
     2461 tagttatcaa catttcacaa ttgaaggaaa taaagttgaa gtactcaaca aaaacttact
     2521 tcagaattaa atttttgggg ggaacatagg catcctatga caggtgacca caagcccctc
     2581 aacgcaatct aatattttac aaagtggtaa aattctttcg ttcttcgttt taatatacag
     2641 tcatttattg attctattac attaatattc ctacgcttcg gctcacataa ttaacaggac
     2701 ttcgagtccg ttaaacttgg gatcaactaa tttctacgga ttagggataa cagggtaatg
     2761 cggccgccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2821 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2881 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2941 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3001 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3061 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3121 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3181 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     3241 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3301 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3361 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3421 actcacgtta agggattttg gtcatgcatt ctaggtgatt atttgccgac taccttggtg
     3481 atctcgcctt tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga
     3541 tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc
     3601 ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact
     3661 gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcacc agcccagtcg
     3721 ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga
     3781 accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct
     3841 tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga
     3901 atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga
     3961 atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca
     4021 ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc
     4081 cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact
     4141 gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca
     4201 actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat aatgtttaac
     4261 tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctccataa catcaaacat
     4321 cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg taccccaaaa
     4381 aaacagtcat aacaagccat gaaaaccgcc actgcgccgt t
//
