LOCUS       Exported                4477 bp DNA     circular SYN 29-MAY-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4477)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4477)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 29, 2024 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Imported using the Genbank importer. File name: 
            pPpT4_Alph_formatiert.gb
FEATURES             Location/Qualifiers
     source          1..4477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          140..1070
                     /mol_type="other DNA"
                     /db_xref="taxon:1182041"
                     /organism="Escherichia coli-Pichia pastoris shuttle vector
                     pPpT4"
     source          2972..3346
                     /mol_type="other DNA"
                     /db_xref="taxon:1182041"
                     /organism="Escherichia coli-Pichia pastoris shuttle vector
                     pPpT4"
     source          3348..3823
                     /mol_type="other DNA"
                     /db_xref="taxon:1182041"
                     /organism="Escherichia coli-Pichia pastoris shuttle vector
                     pPpT4"
     misc_feature    6..131
                     /label=P_AOX1_Syn_dBamHI
     regulatory      140..1070
                     /label=AOX1 promoter
                     /note="AOX1 promoter"
                     /regulatory_class="promoter"
     primer_bind     complement(1071..1083)
                     /label=rv(pPpT4)
     CDS             1084..1350
                     /codon_start=1
                     /label=Signal sequence (MF-alpha)
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
     CDS             1351..2067
                     /codon_start=1
                     /label=Gene of Interest GOI (eGFP)
                     /translation="ASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     2068..2087
                     /label=fw(pPpT4)
     terminator      2088..2334
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2349..2906
                     /label=P_ILV5
     promoter        2907..2970
                     /label=P_EM72_Syn
     CDS             2972..3346
                     /codon_start=1
                     /product="zeocin resistance protein"
                     /label=zeocin resistance protein
                     /protein_id="AFJ20653.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      3348..3823
                     /label=AOD terminator
                     /note="AOD terminator"
                     /regulatory_class="terminator"
     rep_origin      3870..4458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gatccaattc ccgctttgac tgcctgaaat ctccatcgcc tacaatgatg acatttggat
       61 ttggttgact catgttggta ttgtgaaata gacgcagatc gggaacactg aaaaatacac
      121 agttattatt catttaaata acatccaaag acgaaaggtt gaatgaaacc tttttgccat
      181 ccgacatcca caggtccatt ctcacacata agtgccaaac gcaacaggag gggatacact
      241 agcagcagac cgttgcaaac gcaggacctc cactcctctt ctcctcaaca cccacttttg
      301 ccatcgaaaa accagcccag ttattgggct tgattggagc tcgctcattc caattccttc
      361 tattaggcta ctaacaccat gactttatta gcctgtctat cctggccccc ctggcgaggt
      421 tcatgtttgt ttatttccga atgcaacaag ctccgcatta cacccgaaca tcactccaga
      481 tgagggcttt ctgagtgtgg ggtcaaatag tttcatgttc cccaaatggc ccaaaactga
      541 cagtttaaac gctgtcttgg aacctaatat gacaaaagcg tgatctcatc caagatgaac
      601 taagtttggt tcgttgaaat gctaacggcc agttggtcaa aaagaaactt ccaaaagtcg
      661 gcataccgtt tgtcttgttt ggtattgatt gacgaatgct caaaaataat ctcattaatg
      721 cttagcgcag tctctctatc gcttctgaac cccggtgcac ctgtgccgaa acgcaaatgg
      781 ggaaacaccc gctttttgga tgattatgca ttgtctccac attgtatgct tccaagattc
      841 tggtgggaat actgctgata gcctaacgtt catgatcaaa atttaactgt tctaacccct
      901 acttgacagc aatatataaa cagaaggaag ctgccctgtc ttaaaccttt ttttttatca
      961 tcattattag cttactttca taattgcgac tggttccaat tgacaagctt ttgattttaa
     1021 cgacttttaa cgacaacttg agaagatcaa aaaacaacta attattgaaa gaattccgaa
     1081 acgatgagat tcccatctat tttcaccgct gtcttgttcg ctgcctcctc tgcattggct
     1141 gcccctgtta acactaccac tgaagacgag actgctcaaa ttccagctga agcagttatc
     1201 ggttactctg accttgaggg tgatttcgac gtcgctgttt tgcctttctc taactccact
     1261 aacaacggtt tgttgttcat taacaccact atcgcttcca ttgctgctaa ggaagagggt
     1321 gtctctctcg agaagagaga ggccgaagct gctagcaaag gagaagaact tttcactgga
     1381 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt
     1441 ggagagggtg aaggtgatgc tacatacgga aagcttaccc ttaaatttat ttgcactact
     1501 ggaaaactac ctgttccatg gccaacactt gtcactactt tgacttatgg tgttcaatgc
     1561 ttttcccgtt atccggatca tatgaaacgg catgactttt tcaagagtgc catgcccgaa
     1621 ggttatgtac aggaacgcac tatatctttc aaagatgacg ggaactacaa gacgcgtgct
     1681 gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg agttaaaagg tattgatttt
     1741 aaagaagatg gaaacattct cggacacaaa cttgagtaca actataactc acacaatgta
     1801 tacatcacgg cagacaaaca aaagaatgga atcaaagcta acttcaaaat tcgtcacaac
     1861 attgaagatg gttccgttca actagcagac cattatcaac aaaatactcc aattggcgat
     1921 ggccctgtcc ttttaccaga caaccattac ctgtcgacac aatctgccct ttcgaaagat
     1981 cccaacgaaa agcgtgacca catggtcctt cttgagtttg taactgctgc tgggattaca
     2041 catggcatgg atgaattgta caagtaatac gtacctaggg cggccgctca agaggatgtc
     2101 agaatgccat ttgcctgaga gatgcaggct tcatttttga tactttttta tttgtaacct
     2161 atatagtata ggattttttt tgtcattttg tttcttctcg tacgagcttg ctcctgatca
     2221 gcctatctcg cagcagatga atatcttgtg gtaggggttt gggaaaatca ttcgagtttg
     2281 atgtttttct tggtatttcc cactcctctt cagagtacag aagattaagt gagaccttcg
     2341 tttgtgcgga tccttcagta atgtcttgtt tcttttgttg cagtggtgag ccattttgac
     2401 ttcgtgaaag tttctttaga atagttgttt ccagaggcca aacattccac ccgtagtaaa
     2461 gtgcaagcgt aggaagacca agactggcat aaatcaggta taagtgtcga gcactggcag
     2521 gtgatcttct gaaagtttct actagcagat aagatccagt agtcatgcat atggcaacaa
     2581 tgtaccgtgt ggatctaaga acgcgtccta ctaaccttcg cattcgttgg tccagtttgt
     2641 tgttatcgat caacgtgaca aggttgtcga ttccgcgtaa gcatgcatac ccaaggacgc
     2701 ctgttgcaat tccaagtgag ccagttccaa caatctttgt aatattagag cacttcattg
     2761 tgttgcgctt gaaagtaaaa tgcgaacaaa ttaagagata atctcgaaac cgcgacttca
     2821 aacgccaata tgatgtgcgg cacacaataa gcgttcatat ccgctgggtg actttctcgc
     2881 tttaaaaaat tatccgaaaa aattttctag agtgttgtta ctttatactt ccggctcgta
     2941 taatacgaca aggtgtaagg aggactaaac catggctaaa ctcacctctg ctgttccagt
     3001 cctgactgct cgtgatgttg ctggtgctgt tgagttctgg actgatagac tcggtttctc
     3061 ccgtgacttc gtagaggacg actttgccgg tgttgtacgt gacgacgtta ccctgttcat
     3121 ctccgcagtt caggaccagg ttgtgccaga caacactctg gcatgggtat gggttcgtgg
     3181 tctggacgaa ctgtacgctg agtggtctga ggtcgtgtct accaacttcc gtgatgcatc
     3241 tggtccagct atgaccgaga tcggtgaaca gccctggggt cgtgagtttg cactgcgtga
     3301 tccagctggt aactgcgtgc atttcgtcgc agaagagcag gactaacaat tgacacctta
     3361 cgattattta gagagtattt attagtttta ttgtatgtat acggatgttt tattatctat
     3421 ttatgccctt atattctgta actatccaaa agtcctatct tatcaagcca gcaatctatg
     3481 tccgcgaacg tcaactaaaa ataagctttt tatgctcttc tctctttttt tcccttcggt
     3541 ataattatac cttgcatcca cagattctcc tgccaaattt tgcataatcc tttacaacat
     3601 ggctatatgg gagcacttag cgccctccaa aacccatatt gcctacgcat gtataggtgt
     3661 tttttccaca atattttctc tgtgctctct ttttattaaa gagaagctct atatcggaga
     3721 agcttctgtg gccgttatat tcggccttat cgtgggacca cattgcctga attggtttgc
     3781 cccggaagat tggggaaact tggatctgat taccttagct gcaggtacca ctgagcgtca
     3841 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3901 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3961 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4021 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4081 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4141 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4201 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4261 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4321 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4381 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4441 gggcggagcc tatggaaaaa cgccagcaac gcggcct
//
