LOCUS       40924_34889        3780 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPL5617_pUG_PuroR, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3780)
  AUTHORS   MacDonald C, Piper RC.
  TITLE     Puromycin- and methotrexate-resistance cassettes and optimized 
            Cre-recombinase expression plasmids for use in yeast
  JOURNAL   Yeast 32 (5), 423-438 (2015)
  PUBMED    25688547
REFERENCE   2  (bases 1 to 3780)
  AUTHORS   MacDonald C, Piper RC.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 3780)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3780)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; 
            date: "2015"; volume: "32"; issue: "5"; pages: "423-438"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-NOV-2014) Molecular Physiology "
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Sequence Builder (Lasergene) v. 8.0.3 
            Sequencing Technology :: Assembled
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    53..86
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        141..484
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             485..1081
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     terminator      1090..1287
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    1331..1364
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(1331..1364)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(1418..1436)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(1694..2282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2456..3313)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3314..3418)
                     /label=AmpR promoter
     promoter        3764..3780
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acaaccctta atataacttc
       61 gtataatgta tgctatacga agttattagg tctagagatc tgtttagctt gcctcgtccc
      121 cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac agtcttgacg
      181 tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg
      241 tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggct
      301 cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg aattgtcccc
      361 acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg ttcttctttc
      421 atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc gaacataaac
      481 aaccatgact gaatataaac caactgttag attggctact agagatgatg ttccaagagc
      541 tgttagaact ttggctgctg ctttcgctga ttatccagct actagacata ctgttgatcc
      601 agatagacat attgaaagag ttactgaatt gcaagaattg ttcttgacta gagttggttt
      661 ggatattggt aaagtttggg ttgctgatga tggtgctgct gttgctgttt ggactactcc
      721 agaatctgtt gaagctggtg ctgttttcgc tgaaattggt ccaagaatgg ctgaattgtc
      781 tggttctaga ttggctgctc aacaacaaat ggaaggtttg ttggctccac atagaccaaa
      841 agaaccagct tggttcttgg ctactgttgg tgtttctcca gatcatcaag gtaaaggttt
      901 gggttctgct gttgttttgc caggtgttga agctgctgaa agagctggtg ttccagcttt
      961 cttggaaact tctgctccaa gaaatttgcc attctatgaa agattgggtt tcactgttac
     1021 tgctgatgtt gaatgtccaa aagatagagc tacttggtgt atgactagaa aaccaggtgc
     1081 ttaaagagta ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt
     1141 ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc
     1201 ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat
     1261 cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg ccatctcgag
     1321 aacccttaat ataacttcgt ataatgtatg ctatacgaag ttattaggtg atatcagatc
     1381 cactagtggc ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt
     1441 cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1501 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1561 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1621 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1681 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1741 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1801 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1861 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     1921 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1981 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2041 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2101 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     2161 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2221 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2281 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2341 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2401 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2461 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2521 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2581 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2641 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2701 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2761 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2821 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2881 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2941 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3001 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3061 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3121 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3181 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3241 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3301 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3361 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3421 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     3481 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     3541 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     3601 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     3661 atgcggcatc agagcagatt gtactgagag tgcaccatat ggacatattg tcgttagaac
     3721 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
//
