LOCUS       sequence-213436-        3788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pPYK1 in domestication vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3788)
  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 3788)
  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-pPYK1 (01-11)
FEATURES             Location/Qualifiers
     source          1..3788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    232..253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(268..273,274..291,292..298)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    306..322
                     /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     311..333
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     330..346
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     330..346
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        364..382
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     364..381
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1249..1258
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1253..1262
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1292..1311)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1293..1311)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1318..1335)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1318..1334)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1327..1349)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1475..1930
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1562..1581)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1772..1793
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2008..2112
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2113..2181,2182..2973)
                     /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(2331..2350)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3144..3732
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3633..3652
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      241 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      301 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      361 gctatttagg tgacactata gaatactcaa gctatgcatc caacgcgttg ggagctctcc
      421 catatggtcg acctgcaggc ggccgcgaat tcactagtga ttgggcgatg agtggtctcg
      481 ggaggaaagt ttttccggca agctaaatgg aaaaaggaaa gattattgaa agagaaagaa
      541 agaaaaaaaa aaaatgtaca cccagacatc gggcttccac aatttcggct ctattgtttt
      601 ccatctctcg caacggcggg attcctctat ggcgtgtgat gtctgtatct gttacttaat
      661 ccagaaactg gcacttgacc caactctgcc acgtgggtcg ttttgccatc gacagattgg
      721 gagattttca tagtagaatt cagcatgata gctacgtaaa tgtgttccgc accgtcacaa
      781 agtgttttct actgttcttt cttctttcgt tcattcagtt gagttgagtg agtgctttgt
      841 tcaatggatc ttagctaaaa tgcatatttt ttctcttggt aaatgaatgc ttgtgatgtc
      901 ttccaagtga tttcctttcc ttcccatatg atgctaggta cctttagtgt cttcctaaaa
      961 aaaaaaaaag gctcgccatc aaaacgatat tcgttggctt ttttttctga attataaata
     1021 ctctttggta acttttcatt tccaagaacc tcttttttcc agttatatca tggtcccctt
     1081 tcaaagttat tctctactct ttttcatatt cattcttttt catcctttgg ttttttattc
     1141 ttaacttgtt tattattctc tcttgtttct atttacaaga caccaatcaa aacaaataaa
     1201 acatcatcac accatcgaga ccactcatcg cccaatcgaa ttcccgcggc cgccatggcg
     1261 gccgggagca tgcgacgtcg ggcccaattc gccctatagt gagtcgtatt acaattcact
     1321 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     1381 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     1441 ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc
     1501 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1561 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1621 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1681 aaacttgatt agggtgatgg ttcacgtagt gggccagcgc cctgatagac ggtttttcgc
     1741 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     1801 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     1861 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     1921 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     1981 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2041 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2101 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2161 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2221 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2281 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2341 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2401 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2461 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2521 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2581 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2641 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2701 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     2761 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     2821 gagcgtgggt cgcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     2881 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2941 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3001 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3061 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3121 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3181 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3241 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3301 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3361 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3421 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3481 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3541 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3601 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3661 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3721 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3781 tttgctca
//
