LOCUS       sequence-198596-        3462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pPGI1 in domestication vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3462)
  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 3462)
  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-PGI1 (01-11)
FEATURES             Location/Qualifiers
     source          1..3462
                     /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        complement(627..645)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(628..645)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(663..679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(663..679)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(676..698)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    687..703
                     /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(711..717,718..735,736..741))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    756..777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(894..911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1065..1653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1145..1164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1824..2615,2616..2684))
                     /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     2447..2466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2685..2789)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2867..3322)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3004..3025)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3216..3235
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(3448..3462,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3462,1..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 gaggtaacaa aaatcacgat ctgggtgggt gtgggtgtat tggattatag gaagccacgc
      181 gctcaacctg gaattacagg aagctggtaa ttttttgggt ttgcaatcat caccatctgc
      241 acgttgttat aatgtcccgt gtctatatat atccattgac ggtattctat ttttttgcta
      301 ttgaaatgag cgttttttgt tactacaatt ggttttacag acggaatttt ccctatttgt
      361 ttcgtcccat ttttcctttt ctcattgttc tcatatctta aaaaggtcct ttcttcataa
      421 tcaatgcttt cttttactta atattttact tgcattcagt gaattttaat acatattcct
      481 ctagtcttgc aaaatcgatt tagaatcaag ataccagcct aaaaccatcg agaccactca
      541 tcgcccaatc actagtgaat tcgcggccgc ctgcaggtcg accatatggg agagctccca
      601 acgcgttgga tgcatagctt gagtattcta tagtgtcacc taaatagctt ggcgtaatca
      661 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
      721 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
      781 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
      841 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
      901 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
      961 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1021 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1081 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1141 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1201 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1261 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1321 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1381 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1441 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1501 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     1561 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     1621 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     1681 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     1741 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     1801 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1861 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     1921 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgcgaccc acgctcaccg
     1981 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2041 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2101 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2161 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2221 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2281 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2341 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2401 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     2461 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2521 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     2581 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     2641 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     2701 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     2761 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgatgcg
     2821 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt
     2881 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2941 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3001 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3061 aaaaccgtct atcagggcgc tggcccacta cgtgaaccat caccctaatc aagttttttg
     3121 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3181 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3241 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3301 aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc
     3361 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     3421 gattaagttg ggtaacgcca gggttttccc agtcacgacg tt
//
