LOCUS       sequence-199845-        4421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid vector pDGB2-Omega-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 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-1
FEATURES             Location/Qualifiers
     source          1..4421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1147..1470)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1152..1171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1365..1387
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1379..1396
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1380..1396
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1397..1453
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1466..1482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1466..1482)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1479..1501)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1490..1506
                     /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(1514..1520,1521..1538,1539..1544))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1559..1580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2697..3285)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2777..2796)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3373..4164)
                     /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 atgtccactg ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg
       61 gtagcagcga agtcgaggca tttctgtcct ggtgggcggc cgctagggat aacagggtaa
      121 tcgagatagt ttgacgttcg ttttttactt tgaatatact cgtagtcttt ttactttttg
      181 agtataaaaa aaaaatgact agcaaaataa aattagtagt ctaaaaaaga aagctcgcac
      241 tcaggatcga actaaggacc aacagatttg caatctgctg cgctaccact gcgccatacg
      301 agcttttgaa ttatggtaat tttgattatc ctagaatgtt atatctcaat atctcaatat
      361 attttggaca tctatgaaac acccataaag cagccgctac caaacagaca agattcagta
      421 tgtaaggtaa ataccttttt gcacagttaa actacccaaa cttattaaag cttgataaat
      481 tactgaaatt ccacctttca gttagattca ggcctcatat agattagata tagggtacgt
      541 aacattctgt caaccaagtt gttggaatga aagtctaaaa tgtcatctat tcggtagcac
      601 tcatgttact agtatactgt cacatgcggt gtaacgtggg gacataaaac agacatcaaa
      661 tataatggaa gctgaaatgc aaagatcgat aatgtaatag gaatgaaaca tataaaacga
      721 aaggagaagt aatggtaata ttagtatgta gaaataccga ttcaattttg gggattctta
      781 tattctcgag agaatttcta gtataatctg tatacataat attataggct ttaccaacaa
      841 tggaatttcg acaattatca tattattcac caattaatca caagttggta atgagtttga
      901 taacaagtta ctttcttaac aacgttagta tcgtcaaaac actcggtttt actcgagctt
      961 gtagcacaat aataccgtgt agagttctgt attgttcttc ttagtgcttg tatatgctca
     1021 tcccgacctt ccattggatc cggtctcagg agtgagacgc acagcttgtc tgtaagcgga
     1081 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     1141 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     1201 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     1261 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     1321 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     1381 taaaacgacg gccagtgaat tcgagctcgg tacccgggga tcctctagag tcgacctgca
     1441 ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     1501 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     1561 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     1621 tgtcgtgcca gctgcgtctc acgctgtcat gagaccggat cctttgcgaa accctatgct
     1681 ctgttgttcg gatttgaaat tttaaaacta cattaatgtg ttagtttttc tttctttctt
     1741 tctttgtctt gacgtgattt ggacttctgt cttgcattcg cgtccattca tctgacccaa
     1801 tattcctttt ggttttgtta tccttataaa aagaaaggaa gcttcttaga gggaaaaaaa
     1861 tgatgaagag taatgccaaa atataaataa ataaataaat atgaaaatca ttttctattt
     1921 ttaatagaat aagaagagca tcttaagatt acaatttcaa gaaatagttt acacagtata
     1981 tccaataact ccaataaact actttcctat acaaatttct atggtgggat taatagtaaa
     2041 acttctgtac ttctctaatt caccaagaaa ttaaggtaaa catctggtaa gcactatcca
     2101 gctttttgct attacacata tggcttttct gcaatcattt cttcccattt tgtctcaagc
     2161 cgttagtctt gaaaccacag gcggagtaga gttacttgat gcggtatttt acatgccttt
     2221 tttcactgca aaaaaaatga aatacatatt tacacgattt gcaggacagt ttacgatagt
     2281 gagtatgcag aatagttaac acctttgttt tatccttttg tgtcttaatt atatgatata
     2341 aaggcgcctg gcgttatcgg atagtaatag atgctagtta tcaacatttc acaattgaag
     2401 gaaataaagt tgaagtactc aacaaaaact tacttcagaa ttaaattttt ggggggaaca
     2461 taggcatcct atgacaggtg accacaagcc cctcaacgca atctaatatt ttacaaagtg
     2521 gtaaaattct ttcgttcttc gttttaatat acagtcattt attgattcta ttacattaat
     2581 attcctacgc ttcggctcac ataattaaca ggacttcgag tccgttaaac ttgggatcaa
     2641 ctaatttcta cggattaggg ataacagggt aatgcggccg ccgcgttgct ggcgtttttc
     2701 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2761 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2821 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2881 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2941 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3001 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3061 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3121 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3181 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3241 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3301 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3361 cattctaggt gattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa
     3421 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg
     3481 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
     3541 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
     3601 aagcactaca tttcgctcat caccagccca gtcgggcggc gagttccata gcgttaaggt
     3661 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
     3721 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
     3781 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
     3841 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
     3901 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
     3961 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
     4021 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
     4081 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
     4141 ttcggcgatc accgcttccc tcataatgtt taactttgtt ttagggcgac tgccctgctg
     4201 cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg
     4261 cttggatgcc cgaggcatag actgtacccc aaaaaaacag tcataacaag ccatgaaaac
     4321 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg
     4381 catacgctac ttgcattaca gcttacgaac cgaacaggct t
//
