LOCUS       sequence-198026-        4043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pPGK1 in domestication vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4043)
  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 4043)
  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-PGK1 (01-11)
FEATURES             Location/Qualifiers
     source          1..4043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1175..1184
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1179..1188
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1218..1237)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1219..1237)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1244..1261)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1244..1260)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1253..1275)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1401..1856
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1488..1507)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1698..1719
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1934..2038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2039..2107,2108..2899)
                     /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     complement(2257..2276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3070..3658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3559..3578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3812..3829
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3946..3967
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3982..3987,3988..4005,4006..4012)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4020..4036
                     /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)."
     primer_bind     join(4025..4043,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agctatttag gtgacactat agaatactca
       61 agctatgcat ccaacgcgtt gggagctctc ccatatggtc gacctgcagg cggccgcgaa
      121 ttcactagtg attgggcgat gagtggtctc gggaggaata ccttcaaaga atggggtctt
      181 atcttgtttt gcaagtacca ctgagcagga taataataga aatgataata tactatagta
      241 gagataacgt cgatgacttc ccatactgta attgctttta gttgtgtatt tttagtgtgc
      301 aagtttctgt aaatcgatta attttttttt ctttcctctt tttattaacc ttaattttta
      361 ttttagattc ctgacttcaa ctcaagacgc acagatatta taacatctgc ataataggca
      421 tttgcaagaa ttactcgtga gtaaggaaag agtgaggaac tatcgcatac ctgcatttaa
      481 agatgccgat ttgggcgcga atcctttatt ttggcttcac cctcatacta ttatcagggc
      541 cagaaaaagg aagtgtttcc ctccttcttg aattgatgtt accctcataa agcacgtggc
      601 ctcttatcga gaaagaaatt accgtcgctc gtgatttgtt tgcaaaaaga acaaaactga
      661 aaaaacccag acacgctcga cttcctgtct tcctattgat tgcagcttcc aatttcgtca
      721 cacaacaagg tcctagcgac ggctcacagg ttttgtaaca agcaatcgaa ggttctggaa
      781 tggcgggaaa gggtttagta ccacatgcta tgatgcccac tgtgatctcc agagcaaagt
      841 tcgttcgatc gtactgttac tctctctctt tcaaacagaa ttgtccgaat cgtgtgacaa
      901 caacagcctg ttctcacaca ctcttttctt ctaaccaagg gggtggttta gtttagtaga
      961 acctcgtgaa acttacattt acatatatat aaacttgcat aaattggtca atgcaagaaa
     1021 tacatatttg gtcttttcta attcgtagtt tttcaagttc ttagatgctt tctttttctc
     1081 ttttttacag atcatcaagg aagtaattat ctacttttta caacaaatat aaaacaacca
     1141 tcgagaccac tcatcgccca atcgaattcc cgcggccgcc atggcggccg ggagcatgcg
     1201 acgtcgggcc caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac
     1261 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     1321 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     1381 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1441 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1501 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1561 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1621 tgatggttca cgtagtgggc cagcgccctg atagacggtt tttcgccctt tgacgttgga
     1681 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1741 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1801 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     1861 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatcag gtggcacttt
     1921 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1981 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     2041 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     2101 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     2161 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     2221 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2281 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2341 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2401 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2461 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2521 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2581 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2641 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2701 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtcgcg
     2761 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2821 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2881 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2941 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3001 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3061 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3121 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3181 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3241 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3301 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3361 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3421 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3481 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3541 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3601 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3661 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3721 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3781 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3841 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     3901 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     3961 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     4021 tgtgagcgga taacaatttc aca
//
