LOCUS       Exported                3598 bp DNA     circular SYN 30-SEP-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pPICZ-alpha C
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3598)
  AUTHORS   11111111
  TITLE     Direct Submission
  JOURNAL   Exported Sep 30, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             356..622
                     /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             695..724
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             740..757
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      837..1083
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1098..1509
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1517..1564
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1583..1957
                     /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      2023..2270
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(2345..2933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        join(3015..3598,1..355)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
ORIGIN
        1 tgcttagcgc agtctctcta tcgcttctga accccggtgc acctgtgccg aaacgcaaat
       61 ggggaaacac ccgctttttg gatgattatg cattgtctcc acattgtatg cttccaagat
      121 tctggtggga atactgctga tagcctaacg ttcatgatca aaatttaact gttctaaccc
      181 ctacttgaca gcaatatata aacagaagga agctgccctg tcttaaacct ttttttttat
      241 catcattatt agcttacttt cataattgcg actggttcca attgacaagc ttttgatttt
      301 aacgactttt aacgacaact tgagaagatc aaaaaacaac taattattcg aaacgatgag
      361 atttccttca atttttactg ctgttttatt cgcagcatcc tccgcattag ctgctccagt
      421 caacactaca acagaagatg aaacggcaca aattccggct gaagctgtca tcggttactc
      481 agatttagaa ggggatttcg atgttgctgt tttgccattt tccaacagca caaataacgg
      541 gttattgttt ataaatacta ctattgccag cattgctgct aaagaagaag gggtatctct
      601 cgagaagaga gaggctgaag catcgatgaa ttcacgtggc ccagccggcc gtctcggatc
      661 ggtacctcga gccgcggcgg ccgccagctt tctagaacaa aaactcatct cagaagagga
      721 tctgaatagc gccgtcgacc atcatcatca tcatcattga gtttgtagcc ttagacatga
      781 ctgttcctca gttcaagttg ggcacttacg agaagaccgg tcttgctaga ttctaatcaa
      841 gaggatgtca gaatgccatt tgcctgagag atgcaggctt catttttgat acttttttat
      901 ttgtaaccta tatagtatag gatttttttt gtcattttgt ttcttctcgt acgagcttgc
      961 tcctgatcag cctatctcgc agctgatgaa tatcttgtgg taggggtttg ggaaaatcat
     1021 tcgagtttga tgtttttctt ggtatttccc actcctcttc agagtacaga agattaagtg
     1081 agaccttcgt ttgtgcggat cccccacaca ccatagcttc aaaatgtttc tactcctttt
     1141 ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa acacccaagc
     1201 acagcatact aaattttccc tctttcttcc tctagggtgt cgttaattac ccgtactaaa
     1261 ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa
     1321 aaatttttat cacgtttctt tttcttgaaa tttttttttt tagttttttt ctctttcagt
     1381 gacctccatt gatatttaag ttaataaacg gtcttcaatt tctcaagttt cagtttcatt
     1441 tttcttgttc tattacaact ttttttactt cttgttcatt agaaagaaag catagcaatc
     1501 taatctaagg ggcggtgttg acaattaatc atcggcatag tatatcggca tagtataata
     1561 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
     1621 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
     1681 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
     1741 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
     1801 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
     1861 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
     1921 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtccgacggc ggcccacggg
     1981 tcccaggcct cggagatccg tccccctttt cctttgtcga tatcatgtaa ttagttatgt
     2041 cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac
     2101 aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat
     2161 ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac
     2221 tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc aagctggaga
     2281 ccaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2341 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2401 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2461 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2521 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2581 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2641 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2701 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2761 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2821 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2881 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2941 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3001 tggtcatgag atcagatcta acatccaaag acgaaaggtt gaatgaaacc tttttgccat
     3061 ccgacatcca caggtccatt ctcacacata agtgccaaac gcaacaggag gggatacact
     3121 agcagcagac cgttgcaaac gcaggacctc cactcctctt ctcctcaaca cccacttttg
     3181 ccatcgaaaa accagcccag ttattgggct tgattggagc tcgctcattc caattccttc
     3241 tattaggcta ctaacaccat gactttatta gcctgtctat cctggccccc ctggcgaggt
     3301 tcatgtttgt ttatttccga atgcaacaag ctccgcatta cacccgaaca tcactccaga
     3361 tgagggcttt ctgagtgtgg ggtcaaatag tttcatgttc cccaaatggc ccaaaactga
     3421 cagtttaaac gctgtcttgg aacctaatat gacaaaagcg tgatctcatc caagatgaac
     3481 taagtttggt tcgttgaaat gctaacggcc agttggtcaa aaagaaactt ccaaaagtcg
     3541 gcataccgtt tgtcttgttt ggtattgatt gacgaatgct caaaaataat ctcattaa
//
