LOCUS       sequence-192046-        3323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  empty BB3 for insertion and overexpression of 1 gene in P. pastoris.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3323)
  AUTHORS   Prielhofer R, Barrero JJ, Steuer S, Gassler T, Zahrl R, Baumann K, 
            Sauer M, Mattanovich D, Gasser B, Marx H
  TITLE     GoldenPiCS: a Golden Gate-derived modular cloning system for applied
            synthetic biology in the yeast Pichia pastoris.
  JOURNAL   BMC Syst Biol. 2017 Dec 8;11(1):123. doi: 10.1186/s12918-017-0492-3.
  PUBMED    29221460
REFERENCE   2  (bases 1 to 3323)
  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 Syst 
            Biol."; date: "2017-12-8"; volume: "11(1):123. doi"; pages: " 
            10.1186/s12918-017-0492-3"
COMMENT     Sequence Label: BB3eH_14*
FEATURES             Location/Qualifiers
     source          1..3323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(62..650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(142..161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     gene            771..2346
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        771..1114
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1115..2143
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1115..2143
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2149..2346
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 cgctgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
       61 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      661 tgatcttttc tacggggtgg agcgcaaaaa accccgcccc tgacagggcg gggttttttc
      721 gcgatcggag gcgtcttcgg atccctcgag gaagacgccg ctccggcatg gacatggagg
      781 cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc
      841 gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata cattttgatg
      901 gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacgtgcgag cagggaaacg
      961 ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag
     1021 gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat
     1081 acagttctca catcacatcc gaacataaac aaccatgggt aaaaagcctg aactcaccgc
     1141 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     1201 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     1261 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     1321 atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac
     1381 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctccctg aaaccgaact
     1441 gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag
     1501 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     1561 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     1621 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     1681 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     1741 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     1801 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     1861 cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg
     1921 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     1981 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     2041 aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     2101 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taatcagtac tgacaataaa
     2161 aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta
     2221 ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat
     2281 gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct
     2341 atactggctt cgctcttaat taagcggccc aaagagtgag aggaaagtac ctgggcaaaa
     2401 tcacacaatt ccaaaccatg ctaaatgaga tttaaagagc aaacgatggc aaaaggcaac
     2461 cgttataaat gtgatctttc ttggcagcta tctgtcaatt tttctaagga acagtgaatt
     2521 catcatagga gagatgttat acgttacata atcatacata ctgcatgtat ctcacctact
     2581 ttacctcatc aactctaaaa cagttctagt cccaacccca gattcctagt catgacacaa
     2641 gtccgcaccg gacaggactc acaaccagca agagaagcta acaaatttac gccccggtaa
     2701 aacattcttt aggggccgtt caatggtaat tttcctctca cccgtttaaa cttacctccg
     2761 ggcggtatct tcaataacct ctgttgtccc cgggtatcat tggaaacagt gagggacgtt
     2821 gaacagaaga gaggatcacc gtaaatttgc cttgcaattg gccctaacca cggatggtta
     2881 acttcaagcc atcacgacag caattgagtc ggcgcatagc taccctcctc ttcttgaccc
     2941 catgcatagg accaacctta accgatggaa caggttcctc cgctccgtcc cctggtagtg
     3001 tctctgcgca agaaatagtt aaggtattaa cactgatctc tcgcaccccc ctcacagtac
     3061 tgttatggtg aattgacaaa gccattggct agattgaaac atgtaattca tatgtaatct
     3121 tgttcaatta acgagcttcg tacagtctca atctagacgt ctgataatgg cgtttgtgct
     3181 cctaatcgat gagccatctc atgtgacgtc tatacgcttc gatggcttcc gtcgcgaata
     3241 tagaaccact tgaaatatgc tgcaaaccac gatccaccct ggtcctgaaa agatataaat
     3301 acagcacatc tagcaggctt ttg
//
