LOCUS       sequence-231147-        4527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Colorize the nuclei.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4527)
  AUTHORS   Barrero JJ, Casler JC, Valero F, Ferrer P, Glick BS
  TITLE     An improved secretion signal enhances the secretion of model 
            proteins from Pichia pastoris.
  JOURNAL   Microb Cell Fact. 2018 Oct 12;17(1):161. doi: 
            10.1186/s12934-018-1009-5.
  PUBMED    30314480
REFERENCE   2  (bases 1 to 4527)
  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: "Microb Cell
            Fact."; date: "2018-10-12"; volume: "17(1):161. doi"; pages: " 
            10.1186/s12934-018-1009-5"
COMMENT     Sequence Label: Htb2-GFP
FEATURES             Location/Qualifiers
     source          1..4527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..483
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     primer_bind     complement(955..983)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     terminator      1718..1964
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1979..2392
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2399..2446
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2465..3493
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KK"
     terminator      3537..3784
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3553..3571)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3859..4447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3939..3958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatcttttt tgtagaaatg tcttggtgtc ctcgtccaat caggtagcca tctctgaaat
       61 atctggctcc gttgcaactc cgaacgacct gctggcaacg taaaattctc cggggtaaaa
      121 cttaaatgtg gagtaatgga accagaaacg tctcttccct tctctctcct tccaccgccc
      181 gttaccgtcc ctaggaaatt ttactctgct ggagagcttc ttctacggcc cccttgcagc
      241 aatgctcttc ccagcattac gttgcgggta aaacggaggt cgtgtacccg acctagcagc
      301 ccagggatgg aaaagtcccg gccgtcgctg gcaataatag cgggcggacg catgtcatga
      361 gattattgga aaccaccaga atcgaatata aaaggcgaac acctttccca attttggttt
      421 ctcctgaccc aaagacttta aatttaattt atttgtccct atttcaatca attgaacaac
      481 tatatggcac caaaagcaga gaagaaacca gcctctaagg ccccagctga gaagaagcct
      541 actgccggaa agaagacttc ttcttcttcc gaaccaaaga agagatctaa ggttagaaag
      601 gagtcttacg cttcctacat ctacaaggtt ttgaagcaga ctcaccctga caccggtatc
      661 tctcagaagg ccatgtctat catgaactct ttcgtcaacg acatcttcga gagaattgct
      721 tctgaagctt ccaagttggc ttcttacaac aagaagtcta ccatctctgc aagagagatc
      781 cagaccgctg ttagactgat tcttccaggt gaattgtcta agcacgccgt ctctgaaggt
      841 actcgtgccg ttaccaaata cacctcatca acccaggcta cggatccagg aagttcgggg
      901 tccagcggtt cttcaggcag ttccatggat agcactgaga gccttttcac tggagttgtc
      961 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
     1021 ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa
     1081 ctacctgttc cttggccaac acttgtcact actttaactt atggtgttca gtgcttttca
     1141 agatacccgg atcatatgaa acagcatgac tttttcaaga gtgccatgcc cgaaggttat
     1201 gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg tgctgaagtc
     1261 aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa
     1321 gatggaaaca ttcttgggca caaattggaa tacaactata actcacacaa tgtatacatc
     1381 atggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattagaca caacattgaa
     1441 gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct
     1501 gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa agatcccaac
     1561 gaaaagagag accacatggt ccttaaagag tttgtaacag ctgctgggat tacacatggc
     1621 agctccggct ccagcgggta agtttgtagc cttagacatg actgttcctc agttcaagtt
     1681 gggcacttac gagaagaccg gtcttgctag attctaatca agaggatgtc agaatgccat
     1741 ttgcctgaga gatgcaggct tcatttttga tactttttta tttgtaacct atatagtata
     1801 ggattttttt tgtcattttg tttcttctcg tacgagcttg ctcctgatca gcctatctcg
     1861 cagctgatga atatcttgtg gtaggggttt gggaaaatca ttcgagtttg atgtttttct
     1921 tggtatttcc cactcctctt cagagtacag aagattaagt gagaccttcg tttgtgcgga
     1981 tcccccacac accatagctt caaaatgttt ctactccttt tttactcttc cagattttct
     2041 cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac taaatttccc
     2101 ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa aagaaaaaag
     2161 agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta tcacgtttct
     2221 ttttcttgaa aaattttttt tttgattttt ttctctttcg atgacctccc attgatattt
     2281 aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg ttctattaca
     2341 acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta agggcggtgt
     2401 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     2461 aaccatggga aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     2521 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     2581 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     2641 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     2701 tgacattggg gagtttagcg agagcctgac ctattgcatc tcccgccgtt cacagggtgt
     2761 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctacaaccgg tcgcggaggc
     2821 tatggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     2881 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     2941 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     3001 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     3061 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     3121 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     3181 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     3241 atcgccacga ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     3301 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     3361 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     3421 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     3481 ggcaaagaaa tagaggcctc gcctcggaga tccgtccccc ttttcctttg tcgatatcat
     3541 gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa
     3601 ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt
     3661 attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc
     3721 atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat
     3781 ttgcaagctg gagaccaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3841 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3901 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3961 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4021 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4081 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4141 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4201 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4261 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4321 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4381 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4441 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4501 acgttaaggg attttggtca tgagatc
//
