LOCUS       40924_26211        6799 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Artificial vector pJL529, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6799)
  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 6799)
  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 6799)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6799)
  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..6799
                     /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..1178
                     /label=Cryptococcus neoformans actin promoter
                     /note="Cryptococcus neoformans actin promoter"
                     /regulatory_class="promoter"
     CDS             1179..2201
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     regulatory      2205..3100
                     /label=Cryptococcus neoformans GAL7 terminator
                     /note="Cryptococcus neoformans GAL7 terminator"
                     /regulatory_class="terminator"
     misc_feature    3101..3162
                     /label=LoxP site
                     /note="LoxP site"
     protein_bind    complement(3121..3154)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(3233..3251)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3258..3274)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3415..3843
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             4187..4978
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4999..5856
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      6030..6618
                     /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 aactcgccca acatgtctat ggaagaagaa ggtctcaccg
     1201 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc
     1261 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc
     1321 tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg
     1381 catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga
     1441 cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac
     1501 tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta
     1561 gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc
     1621 gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg
     1681 acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact
     1741 gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca
     1801 atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg
     1861 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct
     1921 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
     1981 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
     2041 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
     2101 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
     2161 gtggaaaccg acgccccagc actcgtccga gggcaaagga atagagtaga tgccgaccga
     2221 acaagagctg atttcgagaa cgcctcagcc agcaactcgc gcgagcctag caaggcaaat
     2281 gcgagagaac ggccttacgc ttggtggcac agttctcgtc cacagttcgc taagctcgct
     2341 cggctgggtc gcgggagggc cggtcgcagt gattcaggcc cttctggatt gtgttggtcc
     2401 ccagggcacg attgtcatgc ccacgcactc gggtgatctg actgatcccg cagattggag
     2461 atcgccgccc gtgcctgccg attgggtgca gatccacaga cgggtgtggt cgccatgatc
     2521 gcgagtcgat agtggctcca agagcgaagc gagcaggact ggcgccggcc aaagcggtcg
     2581 acagtgctcc gagaacgggt gcgcatagaa attgcatcaa cgcatatagc gctagcagca
     2641 cgccatagtg actggcgatg ctgtcggaat ggacgatcga ccgcaaaccg aagtcggcgg
     2701 cttttctgct gcaaaaacgc tggactggca tgaacttcgg tgaaaaaccg cagcaggagg
     2761 caaacaatga agatctggcg cgccactcag ttgttgtctc gaagcatgta ctgtagtttc
     2821 cagtcttcat tcaagttcaa agatccgtac tctgtgcagc tggttcctgc tggtattggc
     2881 tttcttctaa atatttgtaa tggcggctgg ccgctgtaac gacattgttt gacagagttt
     2941 cataacaagt tgatagctaa cggtgcctct cctacgtaga acagattgca tcttggttaa
     3001 aaaagaaaca gcgttttaat cctgtgcatc tgcaaagacg aaacaggtcc atctttgtct
     3061 gcagtaccag gtttgcacat cacatggttg cgcggatccg agctctcgag aacccttaat
     3121 ataacttcgt ataatgtatg ctatacgaag ttattaggta gtaagggcga attctgcaga
     3181 tatccatcac actggcggcc gctcgagcat gcatctagag ggcccaattc gccctatagt
     3241 gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     3301 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     3361 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc
     3421 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3481 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3541 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3601 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3661 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3721 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3781 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3841 attttaacaa aattcagggc gcaagggctg ctaaaggaag cggaacacgt agaaagccag
     3901 tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct ggacaaggga
     3961 aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc gatagctaga
     4021 ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc cctctggtaa
     4081 ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg
     4141 caggggatca agatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4201 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4261 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4321 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4381 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4441 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4501 ccaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4561 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4621 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4681 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4741 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4801 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4861 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4921 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4981 aattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5041 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5101 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5161 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5221 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5281 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5341 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5401 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5461 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5521 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5581 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5641 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5701 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5761 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5821 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5881 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5941 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6001 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6061 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6661 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6721 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6781 gtgagcgagg aagcggaag
//
