LOCUS       sequence-173370-        1920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type 3a, alpha-mating factor no EAEA - secretion signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1920)
  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 1920)
  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: pPTK006-3a-alpha-MF no EAEA
FEATURES             Location/Qualifiers
     source          1..1920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..57,58..255)
                     /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."
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKR"
     terminator      complement(275..309)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(380..1039)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     954..973
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1221..1809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1710..1729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgagatttc cttcaatttt tactgctgtt ttattcgcag catcctccgc attagctgct
       61 ccagtcaaca ctacaacaga agatgaaacg gcacaaattc cggctgaagc tgtcatcggt
      121 tactcagatt tagaagggga tttcgatgtt gctgttttgc cattttccaa cagcacaaat
      181 aacgggttat tgtttataaa tactactatt gccagcattg ctgctaaaga agaaggggta
      241 tctctcgaga aaagaggttc ttgagaccag accaataaaa aacgcccggc ggcaaccgag
      301 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
      361 aagcgagctc gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
      421 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
      481 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
      541 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
      601 ctgaaacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
      661 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
      721 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
      781 tatcccatat caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca
      841 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
      901 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
      961 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     1021 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     1081 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgcccga
     1141 tcaatcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1201 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1261 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1321 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1381 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1441 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1501 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1561 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1621 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1681 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     1741 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     1801 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     1861 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtag tcggtctcat
//
