LOCUS       sequence-191930-        2506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB1 with insert: promoter PTEF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2506)
  AUTHORS   Prielhofer R, Barrero JJ, Steuer S, Gassler T, Zahrl R, Baumann K, 
            Sauer M, Mattanovich D, Gasser B, Marx H
  TITLE     GoldenPiCS: a Golden Gate-derived modular cloning system for applied
            synthetic biology in the yeast Pichia pastoris.
  JOURNAL   BMC Syst Biol. 2017 Dec 8;11(1):123. doi: 10.1186/s12918-017-0492-3.
  PUBMED    29221460
REFERENCE   2  (bases 1 to 2506)
  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: "BMC Syst 
            Biol."; date: "2017-12-8"; volume: "11(1):123. doi"; pages: " 
            10.1186/s12918-017-0492-3"
COMMENT     Sequence Label: BB1_12_pTEF2
FEATURES             Location/Qualifiers
     source          1..2506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(802..818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(802..818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      866..897
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        898..1000
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1001..1816
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1074..1093)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1840..1867
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1879..2466)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1959..1978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(2506,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagttta tctagtcagc ttgattctag ctgatcgtgg accggaaggt
       61 gagccagtga gttgattgca gtccagttac gctggagtct gaggctcgtc ctgaatgata
      121 tgcggcctcg aattcgaaga cgcggagcgg tactaccatt gacactacac atccttaatt
      181 ccaatcctgt ctggcctcct tcacctttta accatcttgc ccattccaac tcgtgtcaga
      241 ttgcgtatca agtgaaaaaa aaaaaatttt aaatctttaa cccaatcagg taataactgt
      301 cgcctctttt atctgccgca ctgcatgagg tgtcccctta gtgggaaaga gtactgagcc
      361 aaccctggag gacagcaagg gaaaaatacc tacaacttgc ttcataatgg tcgtaaaaac
      421 aatccttgtc ggatataagt gttgtagact gtcccttatc ctctgcgatg ttcttcctct
      481 caaagtttgc gatttctctc tatcagaatt gccatcaaga gactcaggac taatttcgca
      541 gtcccacacg cactcgtaca tgattggctg aaatttccct aaagaatttc tttttcacga
      601 aaattttttt tttacacaag attttcagca gatataaaat ggagagcagg acctccgctg
      661 tgactcttct tttttttctt ttattctcac tacatacatt ttagttattc gccaacatgg
      721 cgtcttcgaa ttcgcgcgtg atcttacggc attatacgta tgatcggtcc acgatcagct
      781 agattatcta gtcagcttga tgtcatagct gtttcctgag gctcaatact gaccatttaa
      841 atcatacctg acctccatag cagaaagtca aaagcctccg accggaggct tttgacttga
      901 tcggcacgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt
      961 ttttgagtta tcgagatttt caggagctaa ggaagctaaa atgagccata ttcaacggga
     1021 aacgtcttgc ttgaagccgc gattaaattc caacatggat gctgatttat atgggtataa
     1081 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc
     1141 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     1201 tgagatggtc aggctaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     1261 tatccgtact cctgatgatg catggttact caccactgcg atcccaggga aaacagcatt
     1321 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     1381 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacggcg atcgcgtatt
     1441 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttggtgcga gtgattttga
     1501 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aactcttgcc
     1561 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     1621 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     1681 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     1741 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc acttgatgct
     1801 cgatgagttt ttctaatgag gacctaaatg taatcacctg gctcaccttc gggtgggcct
     1861 ttctgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1921 atgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1981 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2041 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2101 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2161 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2221 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2281 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2341 tctgctgaag ccagttacct cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2401 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2461 tctcaagaag atcctttgat tttctaccga agaaaggccc acccgt
//
