LOCUS       sequence-199846-        4500 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 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-1
FEATURES             Location/Qualifiers
     source          1..4500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1344..1667)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1349..1368
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1562..1584
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1576..1593
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1577..1593
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1594..1650
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1663..1679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1663..1679)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1676..1698)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1687..1703
                     /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(1711..1717,1718..1735,1736..1741))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1756..1777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2862..3450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2942..2961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3538..4329)
                     /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 gggtaatacc cactgccctg aagtccagca ggtactgagg
      241 cttggatctc aaaagcgtgg ggtgcgtggg cgtgcgctgc tcgaggcatt ttagccgctg
      301 atagtgctgg ctccaggggc cactgcgctt gccatacccg cacacggggt tctttgaggc
      361 acttaattga ccggagattt aaacgatatt gtcgtcgggg attagaagac attactggtg
      421 gggaaccctt gatgataata ggaaatatat gcgcagtatg ctccatcgtt acgagatgtt
      481 agcagcctca gagcatccta atgggaatat caatgcatac acaccggagc ttggatatga
      541 taaacgaaat attcttgaat cgtgagatcg cctgttttca aaaccgttgg aggcagaaac
      601 aattttgtca caagatgggc attctacccc atccttgctg tattattgta gtctcgcttt
      661 cttttatgct ggacaaatga gactactgca catttttata cgttcttggt tttttttaaa
      721 ggtgtggttt cggcattatc ctgccgcacg tttcttggat aattcatcct gattctctat
      781 tttaaacgct tcagcctatc aggatttggt tttgatacat actgcaagag tgtatctcgg
      841 gaacagtcat ttattccgca acaaacttaa ttgcggaacg cgttaggcga tttctagcat
      901 atatcaaata ccgttcgcga tttcttctgg gttcgtctat tttcttttaa atacttatta
      961 acgtactcaa acaactacac ttcgttgtat ctcagaatga gatccctcag tatgacaata
     1021 catcattcta aacgttcgta aaacacatat gaaacaactt tataacaaag cgaacaaaat
     1081 gggcaacatg agatgaaact ccgcgtccct tagctgaact acccaaacgt acgaatgcct
     1141 gaacaattag tttagatccg agattccgcg cttccatcat ttagtataat ccatatttta
     1201 tataatatat aggataagta acagcccgcg aaggatccgg tctcaggagt gagacgcaca
     1261 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     1321 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     1381 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     1441 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     1501 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     1561 tcccagtcac gacgttgtaa aacgacggcc agtgaattcg agctcggtac ccggggatcc
     1621 tctagagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     1681 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     1741 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     1801 tttccagtcg ggaaacctgt cgtgccagct gcgtctcacg ctgtcatgag accggatccg
     1861 gaccaactat catccgctaa ttactgacat taccaaatga gatctgtgaa tgggcaagat
     1921 aaaaaacaaa aattgaaatg tttgacgtta tgtaaaacta ttaattcctt cgctttcggc
     1981 ggtcacagaa tttgcgtgta gctgactctt gttcaatcaa tatcatttgt tactttattt
     2041 gaaagtctgt attactgcgc ctattgtcat ccgtaccaaa gaacgtcaaa aagaaacaag
     2101 ataatttttg tgcttacacc atttatagat cactgagccc agaatatcgc tggagctcag
     2161 tgtaagtggc atgaacacaa ctctgactga tcgcacatat tgccgttatc ataaatacta
     2221 gttgtacttg tcaatgcgac gaatggcatc atgcctatta ttacgttcct ctttttccgt
     2281 ttcatgtttc cagaatgcta ttgaatctaa cacttcaatt ataaaaaaga ataaatccgc
     2341 aataatttta ggctaattgt tgtactgtca agcgaaccta atggttaaaa ttcagaggaa
     2401 ccttcgacgt agtctgatcg ctacttctat atcttatgtt cccagtcaat caaaagttga
     2461 tactataata gctgccattt atacctgtta gttatggcga tcgtttatca cgacaagccc
     2521 aattttggtc gagatgcagc cagttgccat ctattgtttc caaagcattc gtttgattcg
     2581 tatatgatag gaaaacaaat taaacgatgt atcaatttgc tcattcatca taaattgtgt
     2641 ttgaggcatt actaaaagag cagcagatat cagtctgttg gaataaaaat caactatcat
     2701 ctactaacta gtatttacgt tactagtata ttatcatata cggtgttaga agatgacgca
     2761 aatgatgaga aatagtcatc taaattagtg gaagctgaaa cgcaaggatt gataatgtaa
     2821 tagggataac agggtaatgc ggccgccgcg ttgctggcgt ttttccatag gctccgcccc
     2881 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2941 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3001 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3061 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3121 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3181 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3241 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3301 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3361 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3421 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3481 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgcattc taggtgatta
     3541 tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc
     3601 tgcgcgcgag gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat
     3661 gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg
     3721 cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg
     3781 ctcatcacca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc
     3841 aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc
     3901 aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg
     3961 ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt
     4021 agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg
     4081 gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg
     4141 ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg
     4201 cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag
     4261 atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc
     4321 ttccctcata atgtttaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct
     4381 gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg
     4441 catagactgt accccaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
//
