LOCUS       Exported                3597 bp DNA     circular SYN 10-SEP-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pPICZ-alpha B
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3597)
  AUTHORS   11111111
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             418..684
                     /codon_start=1
                     /gene="MF-alpha-1"
                     /product="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /label=alpha-factor secretion signal
                     /note="Cleavage by the Kex2 protease occurs after the 
                     dibasic KR sequence. The EA dipeptides are then removed by 
                     dipeptidyl aminopeptidase A."
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
     CDS             756..785
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             801..818
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      898..1144
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1159..1570
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1578..1625
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1644..2018
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      2084..2331
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(2406..2994)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        join(3076..3597,1..417)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
ORIGIN
        1 gtcggcatac cgtttgtctt gtttggtatt gattgacgaa tgctcaaaaa taatctcatt
       61 aatgcttagc gcagtctctc tatcgcttct gaaccccggt gcacctgtgc cgaaacgcaa
      121 atggggaaac acccgctttt tggatgatta tgcattgtct ccacattgta tgcttccaag
      181 attctggtgg gaatactgct gatagcctaa cgttcatgat caaaatttaa ctgttctaac
      241 ccctacttga cagcaatata taaacagaag gaagctgccc tgtcttaaac cttttttttt
      301 atcatcatta ttagcttact ttcataattg cgactggttc caattgacaa gcttttgatt
      361 ttaacgactt ttaacgacaa cttgagaaga tcaaaaaaca actaattatt cgaaacgatg
      421 agatttcctt caatttttac tgctgtttta ttcgcagcat cctccgcatt agctgctcca
      481 gtcaacacta caacagaaga tgaaacggca caaattccgg ctgaagctgt catcggttac
      541 tcagatttag aaggggattt cgatgttgct gttttgccat tttccaacag cacaaataac
      601 gggttattgt ttataaatac tactattgcc agcattgctg ctaaagaaga aggggtatct
      661 ctcgagaaaa gagaggctga agctgcagga attcacgtgg cccagccggc cgtctcggat
      721 cggtacctcg agccgcggcg gccgccagct ttctagaaca aaaactcatc tcagaagagg
      781 atctgaatag cgccgtcgac catcatcatc atcatcattg agtttgtagc cttagacatg
      841 actgttcctc agttcaagtt gggcacttac gagaagaccg gtcttgctag attctaatca
      901 agaggatgtc agaatgccat ttgcctgaga gatgcaggct tcatttttga tactttttta
      961 tttgtaacct atatagtata ggattttttt tgtcattttg tttcttctcg tacgagcttg
     1021 ctcctgatca gcctatctcg cagctgatga atatcttgtg gtaggggttt gggaaaatca
     1081 ttcgagtttg atgtttttct tggtatttcc cactcctctt cagagtacag aagattaagt
     1141 gagaccttcg tttgtgcgga tcccccacac accatagctt caaaatgttt ctactccttt
     1201 tttactcttc cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag
     1261 cacagcatac taaattttcc ctctttcttc ctctagggtg tcgttaatta cccgtactaa
     1321 aggtttggaa aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata
     1381 aaaattttta tcacgtttct ttttcttgaa attttttttt ttagtttttt tctctttcag
     1441 tgacctccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt tcagtttcat
     1501 ttttcttgtt ctattacaac tttttttact tcttgttcat tagaaagaaa gcatagcaat
     1561 ctaatctaag gggcggtgtt gacaattaat catcggcata gtatatcggc atagtataat
     1621 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
     1681 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact
     1741 tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg
     1801 tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg
     1861 agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg
     1921 ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
     1981 gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtccgacgg cggcccacgg
     2041 gtcccaggcc tcggagatcc gtcccccttt tcctttgtcg atatcatgta attagttatg
     2101 tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga
     2161 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
     2221 tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata
     2281 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caagctggag
     2341 accaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2401 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2461 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2521 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2581 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2641 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2701 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2761 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2821 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     2881 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2941 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3001 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3061 ttggtcatga gatcagatct aacatccaaa gacgaaaggt tgaatgaaac ctttttgcca
     3121 tccgacatcc acaggtccat tctcacacat aagtgccaaa cgcaacagga ggggatacac
     3181 tagcagcaga ccgttgcaaa cgcaggacct ccactcctct tctcctcaac acccactttt
     3241 gccatcgaaa aaccagccca gttattgggc ttgattggag ctcgctcatt ccaattcctt
     3301 ctattaggct actaacacca tgactttatt agcctgtcta tcctggcccc cctggcgagg
     3361 ttcatgtttg tttatttccg aatgcaacaa gctccgcatt acacccgaac atcactccag
     3421 atgagggctt tctgagtgtg gggtcaaata gtttcatgtt ccccaaatgg cccaaaactg
     3481 acagtttaaa cgctgtcttg gaacctaata tgacaaaagc gtgatctcat ccaagatgaa
     3541 ctaagtttgg ttcgttgaaa tgctaacggc cagttggtca aaaagaaact tccaaaa
//
