LOCUS       sequence-199824-        3517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGAL1 in domestication vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3517)
  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 3517)
  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: pUPD-pGAL1 (01-11)
FEATURES             Location/Qualifiers
     source          1..3517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      73..82
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      77..86
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        128..569
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    128..245
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     540..563
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     promoter        complement(682..700)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(683..700)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(718..734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(718..734)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(731..753)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    742..758
                     /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(766..772,773..790,791..796))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    811..832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(949..966)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1120..1708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1200..1219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1879..2670,2671..2739))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2502..2521
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2740..2844)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2922..3377)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3059..3080)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3271..3290
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(3503..3517,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3517,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg ggcccgacgt
       61 cgcatgctcc cggccgccat ggcggccgcg ggaattcgat tgggcgatga gtggtctcgg
      121 gagagtacgg attagaagcc gccgagcggg tgacagccct ccgaaggaag actctcctcc
      181 gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg
      241 ctccgaacaa taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc
      301 agtaacctgg ccccacaaac cttcaaatga acgaatcaaa ttaacaacca taggatgata
      361 atgcgattag ttttttagcc ttatttctgg ggtaattaat cagcgaagcg gtgatttttg
      421 atctattaac agatatataa atgcaaaaac tgcataacca ctttaactaa tactttcaac
      481 attttcggtt tgtattactt cttattcaaa tgtaataaaa gtatcaacaa aaaattgtta
      541 atatacctct atactttaac gtcaaggaga aaaaacccac catcgagacc actcatcgcc
      601 caatcactag tgaattcgcg gccgcctgca ggtcgaccat atgggagagc tcccaacgcg
      661 ttggatgcat agcttgagta ttctatagtg tcacctaaat agcttggcgt aatcatggtc
      721 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
      781 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
      841 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
      901 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
      961 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1021 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1081 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1141 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1201 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1261 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1321 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1381 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1441 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1501 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1561 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1621 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1681 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1741 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1801 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1861 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1921 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     1981 gggcttacca tctggcccca gtgctgcaat gataccgcgc gacccacgct caccggctcc
     2041 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2101 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2161 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2221 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2281 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2341 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2401 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2461 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2521 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2581 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2641 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2701 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     2761 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     2821 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg
     2881 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     2941 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3001 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3061 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3121 cgtctatcag ggcgctggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3181 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3241 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3301 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3361 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     3421 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     3481 agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
