LOCUS       sequence-199844-        4500 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 YORW-Delta-22 solo LTR loci.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4500)
  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 4500)
  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: YORW-Delta-22-Omega-2
FEATURES             Location/Qualifiers
     source          1..4500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1359..1682)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1364..1383
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1577..1599
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1591..1608
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1592..1608
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1609..1665
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1678..1694)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1678..1694)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1691..1713)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1702..1718
                     /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(1726..1732,1733..1750,1751..1756))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1771..1792
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2873..3461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2953..2972)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3549..4340)
                     /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 actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata
       61 cgctacttgc attacagctt acgaaccgaa caggcttatg tccactgggt tcgtgccttc
      121 atccgtttcc acggtgtgcg tcacccggca accttgggta gcagcgaagt cgaggcattt
      181 ctgtcctggt gggcggccgc tagggataac agggtaatac ccactgccct gaagtccagc
      241 aggtactgag gcttggatct caaaagcgtg gggtgcgtgg gcgtgcgctg ctcgaggcat
      301 tttagccgct gatagtgctg gctccagggg ccactgcgct tgccataccc gcacacgggg
      361 ttctttgagg cacttaattg accggagatt taaacgatat tgtcgtcggg gattagaaga
      421 cattactggt ggggaaccct tgatgataat aggaaatata tgcgcagtat gctccatcgt
      481 tacgagatgt tagcagcctc agagcatcct aatgggaata tcaatgcata cacaccggag
      541 cttggatatg ataaacgaaa tattcttgaa tcgtgagatc gcctgttttc aaaaccgttg
      601 gaggcagaaa caattttgtc acaagatggg cattctaccc catccttgct gtattattgt
      661 agtctcgctt tcttttatgc tggacaaatg agactactgc acatttttat acgttcttgg
      721 ttttttttaa aggtgtggtt tcggcattat cctgccgcac gtttcttgga taattcatcc
      781 tgattctcta ttttaaacgc ttcagcctat caggatttgg ttttgataca tactgcaaga
      841 gtgtatctcg ggaacagtca tttattccgc aacaaactta attgcggaac gcgttaggcg
      901 atttctagca tatatcaaat accgttcgcg atttcttctg ggttcgtcta ttttctttta
      961 aatacttatt aacgtactca aacaactaca cttcgttgta tctcagaatg agatccctca
     1021 gtatgacaat acatcattct aaacgttcgt aaaacacata tgaaacaact ttataacaaa
     1081 gcgaacaaaa tgggcaacat gagatgaaac tccgcgtccc ttagctgaac tacccaaacg
     1141 tacgaatgcc tgaacaatta gtttagatcc gagattccgc gcttccatca tttagtataa
     1201 tccatatttt atataatata taggataagt aacagcccgc gaagatatcg gtctcagtca
     1261 ggagtgagac gcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     1321 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     1381 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     1441 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     1501 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     1561 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcgagctc
     1621 ggtacccggg gatcctctag agtcgacctg caggcatgca agcttggcgt aatcatggtc
     1681 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1741 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1801 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcgtc tcacgcttga
     1861 gaccgatatc ggaccaacta tcatccgcta attactgaca ttaccaaatg agatctgtga
     1921 atgggcaaga taaaaaacaa aaattgaaat gtttgacgtt atgtaaaact attaattcct
     1981 tcgctttcgg cggtcacaga atttgcgtgt agctgactct tgttcaatca atatcatttg
     2041 ttactttatt tgaaagtctg tattactgcg cctattgtca tccgtaccaa agaacgtcaa
     2101 aaagaaacaa gataattttt gtgcttacac catttataga tcactgagcc cagaatatcg
     2161 ctggagctca gtgtaagtgg catgaacaca actctgactg atcgcacata ttgccgttat
     2221 cataaatact agttgtactt gtcaatgcga cgaatggcat catgcctatt attacgttcc
     2281 tctttttccg tttcatgttt ccagaatgct attgaatcta acacttcaat tataaaaaag
     2341 aataaatccg caataatttt aggctaattg ttgtactgtc aagcgaacct aatggttaaa
     2401 attcagagga accttcgacg tagtctgatc gctacttcta tatcttatgt tcccagtcaa
     2461 tcaaaagttg atactataat agctgccatt tatacctgtt agttatggcg atcgtttatc
     2521 acgacaagcc caattttggt cgagatgcag ccagttgcca tctattgttt ccaaagcatt
     2581 cgtttgattc gtatatgata ggaaaacaaa ttaaacgatg tatcaatttg ctcattcatc
     2641 ataaattgtg tttgaggcat tactaaaaga gcagcagata tcagtctgtt ggaataaaaa
     2701 tcaactatca tctactaact agtatttacg ttactagtat attatcatat acggtgttag
     2761 aagatgacgc aaatgatgag aaatagtcat ctaaattagt ggaagctgaa acgcaaggat
     2821 tgataatgta atagggataa cagggtaatg cggccgccgc gttgctggcg tttttccata
     2881 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2941 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3001 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3061 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3121 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3181 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3241 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3301 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3361 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3421 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3481 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgcatt
     3541 ctaggtgatt atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct
     3601 tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta
     3661 gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg
     3721 acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc
     3781 actacatttc gctcatcacc agcccagtcg ggcggcgagt tccatagcgt taaggtttca
     3841 tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga
     3901 cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg
     3961 atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc
     4021 agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact
     4081 tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg
     4141 atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata
     4201 tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac
     4261 gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg
     4321 gcgatcaccg cttccctcat aatgtttaac tttgttttag ggcgactgcc ctgctgcgta
     4381 acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg
     4441 gatgcccgag gcatagactg taccccaaaa aaacagtcat aacaagccat gaaaaccgcc
//
