LOCUS       sequence-173355-        1896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type 3a, alpha-Amylase followed by alpha-MF-Delta - secretion 
            signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1896)
  AUTHORS   Obst U, Lu TK, Sieber V
  TITLE     A Modular Toolkit for Generating Pichia pastoris Secretion 
            Libraries.
  JOURNAL   ACS Synth Biol. 2017 Mar 15. doi: 10.1021/acssynbio.6b00337.
  PUBMED    28252957
REFERENCE   2  (bases 1 to 1896)
  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: "ACS Synth 
            Biol. 2017 Mar 15. doi: 10.1021/acssynbio.6b00337."
COMMENT     Sequence Label: pPTK009-3a-alpha-Amylase-alpha-MF-Delta
FEATURES             Location/Qualifiers
     source          1..1896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     sig_peptide     55..114
                     /codon_start=1
                     /product="signal sequence of alpha-amylase from Aspergillus
                     niger"
                     /label=alpha-amylase signal sequence
                     /translation="MVAWWSLFLYGLQVAAPALA"
     terminator      complement(305..339)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(410..1069)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     984..1003
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1251..1839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1740..1759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgttctttc ctgcgttatc ccctgattct gtggataacc gtagtcggtc tcatatggtg
       61 gcatggtggt ccttattctt atatggtctt caagttgctg ctcctgccct tgctatgaga
      121 tttcctagta ttttcactgc tgtgctattt gccgctagtt ccgctctagc tgctccagtt
      181 aatactacta ctgaagatga attggagggt gacttcgatg ttgctgttct gcctttttcc
      241 gcttctatcg cagccaagga agaaggtgta tctctagaga agcgtggttc ttgagaccag
      301 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
      361 ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc
      421 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
      481 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
      541 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
      601 caaaactggt gaaactcacc cagggattgg ctgaaacgaa aaacatattc tcaataaacc
      661 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
      721 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
      781 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
      841 ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
      901 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
      961 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     1021 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     1081 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     1141 ggtgaaagtt ggaacctctt acgtgcccga tcaatcatga ccaaaatccc ttaacgtgag
     1201 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1261 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1321 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1381 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     1441 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1501 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1561 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1621 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1681 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1741 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1801 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1861 ttacggttcc tggccttttg ctggcctttt gctcac
//
