LOCUS       sequence-201702-        4438 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 YPRC-Delta-15 solo LTR loci.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4438)
  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 4438)
  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-alpha-2
FEATURES             Location/Qualifiers
     source          1..4438
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(14..829)
                     /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(14..829)
                     /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     737..756
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(1970..2293)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1975..1994
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2188..2210
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2202..2219
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2203..2219
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2220..2276
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2289..2305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2289..2305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2302..2324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2313..2329
                     /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(2337..2343,2344..2361,2362..2367))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2382..2403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3517..4105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3597..3616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcattctagg tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
       61 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
      121 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
      181 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
      241 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
      301 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
      361 tcattcgtga ttgcgcctga gcgagtcgaa atacgcgatc gctgttaaaa ggacaattac
      421 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
      481 ctgaatcagg atattcttct aatacctgga atgctgtttt ccctgggatc gcagtggtga
      541 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
      601 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
      661 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcggtag attgtcgcac
      721 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
      781 aatttaatcg cggccttgag caagacgttt cccgttgaat atggctcata acagaactta
      841 ttatttcctt cctcttttct acagtattta aagatacccc aagaagctaa ttataacaag
      901 acgaactcca attcgcggcc gctagggata acagggtaat cgagatagtt tgacgttcgt
      961 tttttacttt gaatatactc gtagtctttt tactttttga gtataaaaaa aaaatgacta
     1021 gcaaaataaa attagtagtc taaaaaagaa agctcgcact caggatcgaa ctaaggacca
     1081 acagatttgc aatctgctgc gctaccactg cgccatacga gcttttgaat tatggtaatt
     1141 ttgattatcc tagaatgtta tatctcaata tctcaatata ttttggacat ctatgaaaca
     1201 cccataaagc agccgctacc aaacagacaa gattcagtat gtaaggtaaa tacctttttg
     1261 cacagttaaa ctacccaaac ttattaaagc ttgataaatt actgaaattc cacctttcag
     1321 ttagattcag gcctcatata gattagatat agggtacgta acattctgtc aaccaagttg
     1381 ttggaatgaa agtctaaaat gtcatctatt cggtagcact catgttacta gtatactgtc
     1441 acatgcggtg taacgtgggg acataaaaca gacatcaaat ataatggaag ctgaaatgca
     1501 aagatcgata atgtaatagg aatgaaacat ataaaacgaa aggagaagta atggtaatat
     1561 tagtatgtag aaataccgat tcaattttgg ggattcttat attctcgaga gaatttctag
     1621 tataatctgt atacataata ttataggctt taccaacaat ggaatttcga caattatcat
     1681 attattcacc aattaatcac aagttggtaa tgagtttgat aacaagttac tttcttaaca
     1741 acgttagtat cgtcaaaaca ctcggtttta ctcgagcttg tagcacaata ataccgtgta
     1801 gagttctgta ttgttcttct tagtgcttgt atatgctcat cccgaccttc cattaagctt
     1861 cgtctcagtc aggagagaga cccacagctt gtctgtaagc ggatgccggg agcagacaag
     1921 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     1981 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     2041 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     2101 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     2161 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2221 aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttggcg
     2281 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2341 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2401 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgggg
     2461 tctctcgctt gagacgaagc tttttgcgaa accctatgct ctgttgttcg gatttgaaat
     2521 tttaaaacta cattaatgtg ttagtttttc tttctttctt tctttgtctt gacgtgattt
     2581 ggacttctgt cttgcattcg cgtccattca tctgacccaa tattcctttt ggttttgtta
     2641 tccttataaa aagaaaggaa gcttcttaga gggaaaaaaa tgatgaagag taatgccaaa
     2701 atataaataa ataaataaat atgaaaatca ttttctattt ttaatagaat aagaagagca
     2761 tcttaagatt acaatttcaa gaaatagttt acacagtata tccaataact ccaataaact
     2821 actttcctat acaaatttct atggtgggat taatagtaaa acttctgtac ttctctaatt
     2881 caccaagaaa ttaaggtaaa catctggtaa gcactatcca gctttttgct attacacata
     2941 tggcttttct gcaatcattt cttcccattt tgtctcaagc cgttagtctt gaaaccacag
     3001 gcggagtaga gttacttgat gcggtatttt acatgccttt tttcactgca aaaaaaatga
     3061 aatacatatt tacacgattt gcaggacagt ttacgatagt gagtatgcag aatagttaac
     3121 acctttgttt tatccttttg tgtcttaatt atatgatata aaggcgcctg gcgttatcgg
     3181 atagtaatag atgctagtta tcaacatttc acaattgaag gaaataaagt tgaagtactc
     3241 aacaaaaact tacttcagaa ttaaattttt ggggggaaca taggcatcct atgacaggtg
     3301 accacaagcc cctcaacgca atctaatatt ttacaaagtg gtaaaattct ttcgttcttc
     3361 gttttaatat acagtcattt attgattcta ttacattaat attcctacgc ttcggctcac
     3421 ataattaaca ggacttcgag tccgttaaac ttgggatcaa ctaatttcta cggattaggg
     3481 ataacagggt aatgcggccg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3541 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3601 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3661 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3721 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3781 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3841 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3901 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     3961 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4021 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4081 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4141 tcagtggaac gaaaactcac gttaagggat tttggtcatg cattctaggt gattagaaaa
     4201 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     4261 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     4321 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     4381 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatc
//
