LOCUS       40924_26201        6612 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Artificial vector pJL517, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6612)
  AUTHORS   Patel RD, Lodge JK, Baker LG.
  TITLE     Going green in Cryptococcus neoformans: the recycling of a 
            selectable drug marker
  JOURNAL   Fungal Genet. Biol. 47 (3), 191-198 (2010)
  PUBMED    19944774
REFERENCE   2  (bases 1 to 6612)
  AUTHORS   Patel RD, Lodge JK, Baker LG.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-FEB-2011) Molecular Microbiology, Washington 
            University, 660 S Euclid Ave, St. Louis, MO 63110, USA
REFERENCE   3  (bases 1 to 6612)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6612)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Fungal 
            Genet. Biol."; date: "2010"; volume: "47"; issue: "3"; pages: 
            "191-198"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-FEB-2011) Molecular Microbiology, Washington University, 660 S 
            Euclid Ave, St. Louis, MO 63110, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    310..343
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      354..1174
                     /label=Cryptococcus neoformans actin promoter
                     /note="Cryptococcus neoformans actin promoter"
                     /regulatory_class="promoter"
     CDS             1176..1967
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      1971..2913
                     /label=Cryptococcus neoformans NMT terminator
                     /note="Cryptococcus neoformans NMT terminator"
                     /regulatory_class="terminator"
     misc_feature    2914..2975
                     /label=LoxP site
                     /note="LoxP site"
     protein_bind    complement(2934..2967)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(3046..3064)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3071..3087)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3228..3656
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             4000..4791
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4812..5669
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5843..6431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgc cctttcgaca
      301 acccttaata taacttcgta taatgtatgc tatacgaagt tattaggtct agaaggatgt
      361 gagctggaga gcggggcacg ggaaggggag gactcacata agcatgcagg attcgagtgg
      421 catggtgtgc gctgagtgta tggttgtcgg aggagaggat gatggtaaca acaatagcag
      481 caacgtcact cgacgcgcgt ccggtgtgcc acacggggta acgccgagtc gccgtcaggg
      541 tcgccgagac cactctcaca gcgtcaccgt tggcaccagc tcagcttaca gcttctatcc
      601 tccgccagca tccacataca tcccctatac cgcatccccc acccactgcc caaggtgagt
      661 catcttcccg cccccttccc ttgcccgcca ctcagtcctc catcctccac taatccacct
      721 tatcgcaccc accgcctatc gcacatccga gcacaatgct gggcctgcca ggggctgcta
      781 gatggtgctc tccccacgct gatctgcatg ccggccattg gatcatgggt gctaggtgct
      841 gggtgctgga tgttggatgc tggatgctgg gtgcacgctt ggtcatttcc ttccaggatt
      901 gacggtcgcc gagaggacga cgtggcgttc gacaacgagg gccgatagca ccgcatcgcc
      961 tcgacctgca tccatctgcc ttgtcctttt ggtggaacaa tccatccgtg ctggtgccac
     1021 acgcatagct ggaagagatg gatgtgcgtt gaacagagct gccgtcagga ctttttggtg
     1081 cacggaccct attgtcctcc caatcttcac cgcgtctcct aatatgcagc ctctttgcta
     1141 attgtctttt tccattagta aactcgcccg gatccatgat tgaacaagat ggattgcacg
     1201 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     1261 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     1321 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     1381 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     1441 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     1501 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     1561 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     1621 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     1681 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     1741 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     1801 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     1861 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     1921 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gaattccccc
     1981 tcaaaccccg tctcttcatt ctctcaattt ctctcgaccg caacaagcgg atagacagtc
     2041 ttcttgacat tcttgataca ttgctaacgc tgtgctatga catgcatatt ttgcccaagt
     2101 acaaaagtgt cactcgacac taattgtaat ttcccattta caacgggaga caagaacctg
     2161 caaacatgga catacacaat gtaaaacatt ccaaggaaaa aaaggtagaa ggagtctgat
     2221 tttacagtcc tgtagcttga atgatgtaca ctaacccaag aatgaacacc gcctctgcca
     2281 acaagagtct ccactgtttc ctcatccagc cgagcatctt gaactgggat tgatcacgcc
     2341 caaaattcaa atcgcgtcag cgccctttat cccggaaacg acgatcaaag cattcgaatg
     2401 gaaaatgaca gcagatggga atcagactta ccgcgggtac atctggtcca aaaggaagcc
     2461 cttcaaaggc aatttcaaac catggcgtgg acggcgattc tagctcgtct tctttatgaa
     2521 gtctcttttt cttcttcttg gacgaggagg aggcagagga gcggcggtgt cttatttgng
     2581 gggccttgtg tgcgtgcttt cttttctttt ggatttttcg tccgcgggac gagtttggtc
     2641 gctctctgta ccatgcttgt gagtttgttt atgcctccgt ttccccttgc ttctagtctc
     2701 ttccttctcc acaacttttc catcttcttc ttcttcttct cctccttttt catcctcatt
     2761 tccatctacc tcttcttcca ctttttttaa actttttcca tcgctatccc ttntggggtt
     2821 ttcccaaccg gttcgggact ctcaaaaccc cagaatcacc agagtcctgc tttcctccct
     2881 cttcaccctc ttgtcggata tcatctgtcc cccagctctc gagaaccctt aatataactt
     2941 cgtataatgt atgctatacg aagttattag gtagtaaggg cgaattctgc agatatccat
     3001 cacactggcg gccgctcgag catgcatcta gagggcccaa ttcgccctat agtgagtcgt
     3061 attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     3121 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3181 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag
     3241 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3301 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3361 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3421 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3481 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3541 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3601 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3661 caaaattcag ggcgcaaggg ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag
     3721 aaacggtgct gaccccggat gaatgtcagc tactgggcta tctggacaag ggaaaacgca
     3781 agcgcaaaga gaaagcaggt agcttgcagt gggcttacat ggcgatagct agactgggcg
     3841 gttttatgga cagcaagcga accggaattg ccagctgggg cgccctctgg taaggttggg
     3901 aagccctgca aagtaaactg gatggctttc ttgccgccaa ggatctgatg gcgcagggga
     3961 tcaagatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4021 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4081 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4141 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4201 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4261 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atcccacctt
     4321 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4381 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4441 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4501 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4561 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4621 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4681 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4741 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaattgaa
     4801 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4861 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4921 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4981 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5041 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5101 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5161 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5221 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5281 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5341 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5401 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5461 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5521 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5581 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5641 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5701 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5761 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5821 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5881 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5941 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6001 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6061 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6121 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6181 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6241 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6301 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6361 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6421 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6481 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6541 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6601 aggaagcgga ag
//
