LOCUS       sequence-192459-        2666 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GoldenBraid vector pDGB2-alpha-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      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2666)
  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 2666)
  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-alpha-2
FEATURES             Location/Qualifiers
     source          1..2666
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(1167..1490)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1172..1191
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1385..1407
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1399..1416
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1400..1416
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1417..1473
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1486..1502)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1486..1502)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1499..1521)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1510..1526
                     /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(1534..1540,1541..1558,1559..1564))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1579..1600
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcggccgcta gggataacag ggtaataccc actgccctga agtccagcag gtactgaggc
       61 ttggatctca aaagcgtggg gtgcgtgggc gtgcgctgct cgaggcattt tagccgctga
      121 tagtgctggc tccaggggcc actgcgcttg ccatacccgc acacggggtt ctttgaggca
      181 cttaattgac cggagattta aacgatattg tcgtcgggga ttagaagaca ttactggtgg
      241 ggaacccttg atgataatag gaaatatatg cgcagtatgc tccatcgtta cgagatgtta
      301 gcagcctcag agcatcctaa tgggaatatc aatgcataca caccggagct tggatatgat
      361 aaacgaaata ttcttgaatc gtgagatcgc ctgttttcaa aaccgttgga ggcagaaaca
      421 attttgtcac aagatgggca ttctacccca tccttgctgt attattgtag tctcgctttc
      481 ttttatgctg gacaaatgag actactgcac atttttatac gttcttggtt ttttttaaag
      541 gtgtggtttc ggcattatcc tgccgcacgt ttcttggata attcatcctg attctctatt
      601 ttaaacgctt cagcctatca ggatttggtt ttgatacata ctgcaagagt gtatctcggg
      661 aacagtcatt tattccgcaa caaacttaat tgcggaacgc gttaggcgat ttctagcata
      721 tatcaaatac cgttcgcgat ttcttctggg ttcgtctatt ttcttttaaa tacttattaa
      781 cgtactcaaa caactacact tcgttgtatc tcagaatgag atccctcagt atgacaatac
      841 atcattctaa acgttcgtaa aacacatatg aaacaacttt ataacaaagc gaacaaaatg
      901 ggcaacatga gatgaaactc cgcgtccctt agctgaacta cccaaacgta cgaatgcctg
      961 aacaattagt ttagatccga gattccgcgc ttccatcatt tagtataatc catattttat
     1021 ataatatata ggataagtaa cagcccgcga aaagcttcgt ctcagtcagg agagagaccc
     1081 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     1141 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     1201 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     1261 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1321 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1381 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcgagctcgg tacccgggga
     1441 tcctctagag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     1501 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     1561 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     1621 cgctttccag tcgggaaacc tgtcgtgcca gctggggtct ctcgcttgag acgaagcttg
     1681 gaccaactat catccgctaa ttactgacat taccaaatga gatctgtgaa tgggcaagat
     1741 aaaaaacaaa aattgaaatg tttgacgtta tgtaaaacta ttaattcctt cgctttcggc
     1801 ggtcacagaa tttgcgtgta gctgactctt gttcaatcaa tatcatttgt tactttattt
     1861 gaaagtctgt attactgcgc ctattgtcat ccgtaccaaa gaacgtcaaa aagaaacaag
     1921 ataatttttg tgcttacacc atttatagat cactgagccc agaatatcgc tggagctcag
     1981 tgtaagtggc atgaacacaa ctctgactga tcgcacatat tgccgttatc ataaatacta
     2041 gttgtacttg tcaatgcgac gaatggcatc atgcctatta ttacgttcct ctttttccgt
     2101 ttcatgtttc cagaatgcta ttgaatctaa cacttcaatt ataaaaaaga ataaatccgc
     2161 aataatttta ggctaattgt tgtactgtca agcgaaccta atggttaaaa ttcagaggaa
     2221 ccttcgacgt agtctgatcg ctacttctat atcttatgtt cccagtcaat caaaagttga
     2281 tactataata gctgccattt atacctgtta gttatggcga tcgtttatca cgacaagccc
     2341 aattttggtc gagatgcagc cagttgccat ctattgtttc caaagcattc gtttgattcg
     2401 tatatgatag gaaaacaaat taaacgatgt atcaatttgc tcattcatca taaattgtgt
     2461 ttgaggcatt actaaaagag cagcagatat cagtctgttg gaataaaaat caactatcat
     2521 ctactaacta gtatttacgt tactagtata ttatcatata cggtgttaga agatgacgca
     2581 aatgatgaga aatagtcatc taaattagtg gaagctgaaa cgcaaggatt gataatgtaa
     2641 tagggataac agggtaatgc ggccgc
//
