LOCUS       62056_9145        6447 bp DNA     circular SYN 14-APR-2019
DEFINITION  Cloning vector pEM003, complete sequence.
ACCESSION   MK425747
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6447)
  AUTHORS   Mancera E, Frazer C, Porman AM, Ruiz-Castro S, Johnson AD, Bennett 
            RJ.
  TITLE     Genetic Modification of Closely Related Candida Species
  JOURNAL   Front Microbiol 10, 357 (2019)
  PUBMED    30941104
REFERENCE   2  (bases 1 to 6447)
  AUTHORS   Mancera E, Frazer C, Porman AM, Ruiz S, Johnson AD, Bennett RJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-JAN-2019) Departmento de Ingenieria Genetica, Centro 
            de Investigacion y de Estudios Avanzados del Instituto Politecnico 
            Nacional, Unidad Irapuato, Km. 9.6 Libramiento Norte Carrera 
            Irapuato-Leon, Irapuato, Guanajuato 36824, Mexico
REFERENCE   3  (bases 1 to 6447)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Microbiol 10, 357 (2019)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-JAN-2019) Departmento de Ingenieria Genetica, Centro de 
            Investigacion y de Estudios Avanzados del Instituto Politecnico 
            Nacional, Unidad Irapuato, Km. 9.6 Libramiento Norte Carrera 
            Irapuato-Leon, Irapuato, Guanajuato 36824, Mexico"
FEATURES             Location/Qualifiers
     source          1..6447
                     /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"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     gene            337..3264
                     /gene="ARG4"
                     /label=ARG4
                     /note="derived from Candida albicans"
     misc_feature    337..1321
                     /gene="ARG4"
                     /label=contains promoter and 5' UTR
                     /note="contains promoter and 5' UTR"
     CDS             1322..2728
                     /codon_start=1
                     /transl_table=11
                     /gene="ARG4"
                     /product="argininosuccinate lyase"
                     /label=ARG4
                     /protein_id="QBY25889.1"
                     /translation="MSQQQDKQPSENKLWGGRFTGATDPLMDLYNASLPYDKVMYDADL
                     TGTKVYTQGLNKLGLITTEELHLIHQGLEQIRQEWHDNKFIIKAGDEDIHTANERRLGE
                     IIGKNISGKVHTGRSRNDQVATDMRIFVRESLLNLSKILHQFITAILERAHKEIDVLMP
                     GYTHLQKAQPIRWAHWLSSYATYFTEDYKRLQEIITRVNQSPLGSGALAGHPYGIDREF
                     LAKGLGFDGVIGNSLTAVSDRDFVVESLFWSTLFMNHISRFSEDLIIYSSGEFGFIKLA
                     DAYSTGSSLMPQKKNPDSLELLRGKSGRVFGQLSGFLMSIKSIPSTYNKDMQEDKEPLF
                     DALTTVEHSILIATGVISTLLIDKQNMEKALTMDMLATDLADYLVRKGVPFRETHHISG
                     ECVRKAEEEKLSGIDQLSFEQFQQIDSRFEKDVMETFDFEASVERRDAIGGTAKSAVLK
                     QLENLKSILS"
     misc_feature    2729..3264
                     /gene="ARG4"
                     /label=contains 3' UTR and terminator
                     /note="contains 3' UTR and terminator"
     promoter        complement(3335..3353)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3360..3376)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3514..3813
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             4165..4956
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             5166..5537
                     /label=BleoR
                     /note="antibiotic-binding protein"
     rep_origin      5678..6266
                     /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 gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gccctttgaa tcgatgcttt ctttactgca
      361 aaattatcaa catttttcga tagttcgtta ttattgtttt gttgcgaaat gtagcaatct
      421 atgctttaca tcctatgtgg tactccgttt gttcctttca tacatgcaaa actacaattg
      481 gtatttctaa ccgatattat caatcctttt gtattatcta ataatgggaa aaaaaaaaga
      541 caggaatcac gtcatcgttg tacatttttt acgtgaacta gaagactaat attcacctac
      601 gctgttcttt gaactcaagt ttacaatcat tttgataatc gattgagatc ggtatggtat
      661 tcttaaattg agattgcctc gtagagaggt taatacttag atgtatcata tttaaagatt
      721 tataggaatc actttttgga tgaatatttt ttcaacaaaa gctgttgcga aaaactatta
      781 aaattaaaat gaaaaatggt ctaactaaat taagatccta atagtaatat ctactagaaa
      841 tatatcaaat caacaataaa caagacccct ttagtaagat ttttcaagag tagtctcaaa
      901 taaaccagaa tgaaggattg aaggttttat tcaataaata attggacaga aagttattgt
      961 acagttttaa caacaacaat ttcaaaattt caatttatga catattgacc gacataattc
     1021 agtcatttct aatattcaaa taacaagaca ccttgtataa atgaatccac aatcgtatat
     1081 gaacaaccaa attttatgac taactttggg tcgtgtactt ttgacgattg agtcattttt
     1141 accaattagg taattgaatt ttgttaattt gtcattaacc gcgctggcca tcccaacaat
     1201 aacaacaatc aatttaaaaa cttgacaagg caaaaaaaaa aaaattttat ctttgaaaaa
     1261 tttttcttcc ttttcttctt ctggtcttta cataacagct atcaagaata atcaattaat
     1321 aatgtcacaa caacaagata aacaacctag tgaaaataaa ttatggggtg gtcgtttcac
     1381 cggtgccact gatccattga tggatttata taatgcttca ttaccttatg ataaagttat
     1441 gtatgatgct gatttaactg gtactaaagt atacacacaa ggattaaaca aattaggatt
     1501 aattaccact gaagaattgc atttgattca tcaaggatta gaacaaatca gacaagaatg
     1561 gcatgataat aaattcatta ttaaggcagg tgatgaggat attcatactg ctaatgaacg
     1621 tcgattagga gaaatcattg ggaaaaacat ttctggtaaa gttcacactg gtagatcaag
     1681 aaatgatcaa gttgctaccg atatgagaat tttcgttcgt gaatcattac ttaatttaag
     1741 taaaatttta catcaattca tcacggcaat tcttgaacgg gcacataaag aaatcgatgt
     1801 attaatgcct ggttatactc atttacaaaa agctcaacct attagatggg cacattggtt
     1861 aagttcatat gcaacttatt tcaccgaaga ttataaacgt ttacaagaaa ttataactcg
     1921 tgttaaccaa tctccattag gatctggtgc tttagcaggt catccttatg gtattgatcg
     1981 tgaattttta gctaaaggat taggatttga tggtgtcatt gggaattctt taactgcagt
     2041 ttcagatcgt gattttgttg ttgaaagttt attttggagt acattattta tgaatcatat
     2101 ttctcgattc tcagaagatc taattattta ttctagtggt gaatttgggt tcataaaatt
     2161 agctgatgct tattctactg gttcttcatt gatgccacaa aagaaaaacc ctgactcatt
     2221 agaattatta cgtgggaaaa gtggtagagt atttggtcaa ttatctggat ttttaatgtc
     2281 gatcaaatct attccatcta cttataataa agatatgcaa gaggataaag aaccattatt
     2341 tgatgcattg actacagtgg aacattcaat tttaattgct actggtgtga tttctacttt
     2401 actgattgat aaacaaaata tggaaaaagc tttaactatg gatatgttgg ctactgattt
     2461 agctgattat ttagttagaa aaggagttcc atttagagaa actcatcata tatctggtga
     2521 atgtgttaga aaagctgaag aagaaaaatt atctggaatt gatcaattat cttttgaaca
     2581 atttcaacaa attgattctc gttttgaaaa agacgttatg gaaacttttg attttgaagc
     2641 tagtgtggaa agaagagatg ctattggtgg tactgctaaa agtgccgttt taaaacaatt
     2701 agaaaattta aaatcaattt taagttaatt tataaatagt catataataa tcacagtatt
     2761 gtttaactta cataaaatat aatcaattaa cacagagata ccttgtactc caatactagt
     2821 gttataatgt ttataacagc agctttcaag aataaggaac taaataaaag cttcttccat
     2881 cacaacacac aacacgtcta tatgccataa atacataacg gtcttcctaa taatatccta
     2941 accatacttg caaatgaaaa atacatcaaa ttcagtcatt agtttttttt cctcttgtta
     3001 ttatcatcct catcatcact atattcctca tccagataaa acacacttcc attgtaaaga
     3061 tcgtctgaat cataatcgga gactacctct tcattttctg aagatttgga tttttcgaaa
     3121 taaggatcaa aatcacgata tacagcttcc aaacgataag aatgattaga tattttcaat
     3181 agagaatcat tgagtctgtt gagctctaat gagagtcctt tgattgcttt caatactttc
     3241 ctcacatcaa taatattcac attcaagggc gaattctgca gatatccatc acactggcgg
     3301 ccgctcgagc atgcatctag agggcccaat tcgccctata gtgagtcgta ttacaattca
     3361 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     3421 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     3481 ccttcccaac agttgcgcag cctatacgta cggcagttta aggtttacac ctataaaaga
     3541 gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac gccggggcga
     3601 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt
     3661 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
     3721 gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
     3781 aaaaacgcca ttaacctgat gttctgggga atataaatgt caggcatgag attatcaaaa
     3841 aggatcttca cctagatcct tttcacgtag aaagccagtc cgcagaaacg gtgctgaccc
     3901 cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag
     3961 caggtagctt gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca
     4021 agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta
     4081 aactggatgg ctttctcgcc gccaaggatc tgatggcgca ggggatcaag ctctgatcaa
     4141 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4201 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4261 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4321 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4381 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4441 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4501 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4561 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4621 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4681 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4741 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4801 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4861 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4921 cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa ttattaacgc ttacaatttc
     4981 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acaggtggca
     5041 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5101 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatagcac gtgaggaggg
     5161 ccaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag
     5221 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg
     5281 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc
     5341 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt
     5401 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg
     5461 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg
     5521 tggccgagga gcaggactga cacgtgctaa aacttcattt ttaatttaaa aggatctagg
     5581 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5641 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5701 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5761 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5821 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5881 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5941 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6001 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6061 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6121 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6181 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6241 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6301 gcttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6361 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     6421 gcgagtcagt gagcgaggaa gcggaag
//
