LOCUS       sequence-199811-        4256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid vector pDGB2-alpha-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 4256)
  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 4256)
  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-1
FEATURES             Location/Qualifiers
     source          1..4256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1247..1570)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1252..1271
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1465..1487
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1479..1496
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1480..1496
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1497..1553
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1566..1582)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1566..1582)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1579..1601)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1590..1606
                     /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(1614..1620,1621..1638,1639..1644))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1659..1680
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2765..3353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2845..2864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3441..4256)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3441..4256)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4164..4183
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 aacagaactt attatttcct tcctcttttc tacagtattt aaagataccc caagaagcta
       61 attataacaa gacgaactcc aattcgcggc cgctagggat aacagggtaa tacccactgc
      121 cctgaagtcc agcaggtact gaggcttgga tctcaaaagc gtggggtgcg tgggcgtgcg
      181 ctgctcgagg cattttagcc gctgatagtg ctggctccag gggccactgc gcttgccata
      241 cccgcacacg gggttctttg aggcacttaa ttgaccggag atttaaacga tattgtcgtc
      301 ggggattaga agacattact ggtggggaac ccttgatgat aataggaaat atatgcgcag
      361 tatgctccat cgttacgaga tgttagcagc ctcagagcat cctaatggga atatcaatgc
      421 atacacaccg gagcttggat atgataaacg aaatattctt gaatcgtgag atcgcctgtt
      481 ttcaaaaccg ttggaggcag aaacaatttt gtcacaagat gggcattcta ccccatcctt
      541 gctgtattat tgtagtctcg ctttctttta tgctggacaa atgagactac tgcacatttt
      601 tatacgttct tggttttttt taaaggtgtg gtttcggcat tatcctgccg cacgtttctt
      661 ggataattca tcctgattct ctattttaaa cgcttcagcc tatcaggatt tggttttgat
      721 acatactgca agagtgtatc tcgggaacag tcatttattc cgcaacaaac ttaattgcgg
      781 aacgcgttag gcgatttcta gcatatatca aataccgttc gcgatttctt ctgggttcgt
      841 ctattttctt ttaaatactt attaacgtac tcaaacaact acacttcgtt gtatctcaga
      901 atgagatccc tcagtatgac aatacatcat tctaaacgtt cgtaaaacac atatgaaaca
      961 actttataac aaagcgaaca aaatgggcaa catgagatga aactccgcgt cccttagctg
     1021 aactacccaa acgtacgaat gcctgaacaa ttagtttaga tccgagattc cgcgcttcca
     1081 tcatttagta taatccatat tttatataat atataggata agtaacagcc cgcgaagaat
     1141 tcgtctcagg agagagaccc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     1201 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     1261 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     1321 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     1381 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     1441 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     1501 tcgagctcgg tacccgggga tcctctagag tcgacctgca ggcatgcaag cttggcgtaa
     1561 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1621 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1681 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctggggtct
     1741 ctcgctgtca tgagacgaat tcggaccaac tatcatccgc taattactga cattaccaaa
     1801 tgagatctgt gaatgggcaa gataaaaaac aaaaattgaa atgtttgacg ttatgtaaaa
     1861 ctattaattc cttcgctttc ggcggtcaca gaatttgcgt gtagctgact cttgttcaat
     1921 caatatcatt tgttacttta tttgaaagtc tgtattactg cgcctattgt catccgtacc
     1981 aaagaacgtc aaaaagaaac aagataattt ttgtgcttac accatttata gatcactgag
     2041 cccagaatat cgctggagct cagtgtaagt ggcatgaaca caactctgac tgatcgcaca
     2101 tattgccgtt atcataaata ctagttgtac ttgtcaatgc gacgaatggc atcatgccta
     2161 ttattacgtt cctctttttc cgtttcatgt ttccagaatg ctattgaatc taacacttca
     2221 attataaaaa agaataaatc cgcaataatt ttaggctaat tgttgtactg tcaagcgaac
     2281 ctaatggtta aaattcagag gaaccttcga cgtagtctga tcgctacttc tatatcttat
     2341 gttcccagtc aatcaaaagt tgatactata atagctgcca tttatacctg ttagttatgg
     2401 cgatcgttta tcacgacaag cccaattttg gtcgagatgc agccagttgc catctattgt
     2461 ttccaaagca ttcgtttgat tcgtatatga taggaaaaca aattaaacga tgtatcaatt
     2521 tgctcattca tcataaattg tgtttgaggc attactaaaa gagcagcaga tatcagtctg
     2581 ttggaataaa aatcaactat catctactaa ctagtattta cgttactagt atattatcat
     2641 atacggtgtt agaagatgac gcaaatgatg agaaatagtc atctaaatta gtggaagctg
     2701 aaacgcaagg attgataatg taatagggat aacagggtaa tgcggccgcc gcgttgctgg
     2761 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2821 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2881 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2941 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3001 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3061 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3121 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3181 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     3241 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3301 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3361 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3421 tggtcatgca ttctaggtga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     3481 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     3541 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     3601 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     3661 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     3721 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     3781 ccgttattca ttcgtgattg cgcctgagcg agtcgaaata cgcgatcgct gttaaaagga
     3841 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     3901 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc tgggatcgca
     3961 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     4021 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     4081 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcggtagatt
     4141 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     4201 atgttggaat ttaatcgcgg ccttgagcaa gacgtttccc gttgaatatg gctcat
//
