LOCUS       Exported                7510 bp DNA     circular SYN 23-DEC-2024
DEFINITION  Cloning vector pSFS2A-mNeonGreen, complete sequence.
ACCESSION   MK431400
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7510)
  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 7510)
  AUTHORS   Mancera E, Frazer C, Porman AM, Ruiz S, Johnson AD, Bennett RJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-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 7510)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7510)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 23, 2024 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front
            Microbiol 10, 357 (2019)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (23-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"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(915..7510,1..914)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(142..246)
                     /label=AmpR promoter
     rep_origin      273..729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     870..886
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        896..914
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(944..977)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     misc_feature    982..1529
                     /label=promoter region from Candida albicans MAL2
                     /note="promoter region from Candida albicans MAL2"
     CDS             1536..2804
                     /label=FLP
                     /note="site-specific recombinase"
     misc_feature    2808..3195
                     /label=terminator region from Candida albicans ACT1
                     /note="terminator region from Candida albicans ACT1"
     misc_feature    3202..3699
                     /label=promoter region from Candida albicans ACT1
                     /note="promoter region from Candida albicans ACT1"
     gene            3700..4926
                     /gene="SAT1"
                     /label=SAT1
     CDS             join(3700..3709,4364..4926)
                     /codon_start=1
                     /transl_table=11
                     /gene="SAT1"
                     /product="streptomycin acetyltransferase"
                     /label=SAT1
                     /note="nourseothricin resistance marker"
                     /protein_id="QBY25799.1"
                     /translation="MDGEEVAALVIDNGSHMKISVIPEQVAETLDAENHFIVREVFDVH
                     LSDQGFELSTRSVSPYRKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEH
                     IVVSHTHRGKGVAHSLIEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDL
                     FTYKTRPQVSNETAMYWYWFSGAQDDA"
     misc_feature    4930..5059
                     /label=terminator region from Candida albicans URA3
                     /note="terminator region from Candida albicans URA3"
     protein_bind    complement(5064..5097)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     primer_bind     complement(5099..5115)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5152..5170)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5191..5207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5215..5231)
                     /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)."
     promoter        complement(5239..5269)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5284..5305)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5593..6181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6401..6503
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             6504..7160
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
ORIGIN
        1 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
       61 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      121 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      181 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      241 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
      301 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
      361 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
      421 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
      481 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
      541 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
      601 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
      661 gttaaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
      721 ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
      781 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
      841 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata
      901 cgactcacta tagggcgaat tgggtaccgg gccccccctc gaggaagttc ctatactttc
      961 tagagaatag gaacttcgga tccaataatg attggtttga tatttttgtc tagtaccatc
     1021 tgtaccatta cacttaaatt atctttatat ctgtctaact cgactgtctg gatttcattg
     1081 atgtagtcgt atgcatcgtt agttccaaaa aatattgtca tcaatttgat attggtttcc
     1141 gactctaaaa tttttggaag aatttgtcta gcgtgctctg agttgtagcc actgaaacca
     1201 cggttaataa catccaattt tcggatatac acattctgta atgctggatg aaagccatac
     1261 tgggtacaac taaactgggt gatggagtca ccgaacaaca caaatttacc gtattccatg
     1321 attgctatgg ttgagaattt tttttttttc ttgtcccacg ccatttttca aattatgcag
     1381 ttgagaatgt tagtttttgt gtacaccccg ttcgctgaat atttcggaat aattcaaaga
     1441 ttggggagtg ggggaggcga tagacgaaga cacggtataa aaatgggcaa aattttcccc
     1501 aactttttgc agtggtttaa ctaataatcg tcgacatgcc acaatttgat atattatgta
     1561 aaacaccacc taaggtgctt gttcgtcagt ttgtggaaag gtttgaaaga ccttcaggtg
     1621 agaaaatagc attatgtgct gctgaactaa cctatttatg ttggatgatt acacataacg
     1681 gaacagcaat caagagagcc acattcatga gctataatac tatcataagc aattcgttga
     1741 gtttcgatat tgtcaataaa tcactccagt ttaaatacaa gacgcaaaaa gcaacaattt
     1801 tggaagcctc attaaagaaa ttgattcctg cttgggaatt tacaattatt ccttactatg
     1861 gacaaaaaca tcaatctgat atcactgata ttgtaagtag tttgcaatta cagttcgaat
     1921 catcggaaga agcagataag ggaaatagcc acagtaaaaa aatgcttaaa gcacttctaa
     1981 gtgagggtga aagcatctgg gagatcactg agaaaatact aaattcgttt gagtatactt
     2041 cgagatttac aaaaacaaaa actttatacc aattcctctt cctagctact ttcatcaatt
     2101 gtggaagatt cagcgatatt aagaacgttg atccgaaatc atttaaatta gtccaaaata
     2161 agtatttggg agtaataatc cagtgtttag tgacagagac aaagacaagc gttagtaggc
     2221 acatatactt ctttagcgca aggggtagga tcgatccact tgtatatttg gatgaatttt
     2281 tgaggaattc tgaaccagtc ctaaaacgag taaataggac cggcaattct tcaagcaata
     2341 aacaggaata ccaattatta aaagataact tagtcagatc gtacaataaa gctttgaaga
     2401 aaaatgcgcc ttattcaatc tttgctataa aaaatggccc aaaatctcac attggaagac
     2461 atttgatgac ctcatttctt tcaatgaagg gcctaacgga gttgactaat gttgtgggaa
     2521 attggagcga taagcgtgct tctgccgtgg ccaggacaac gtatactcat cagataacag
     2581 caatacctga tcactacttc gcactagttt ctcggtacta tgcatatgat ccaatatcaa
     2641 aggaaatgat agcattgaag gatgagacta atccaattga ggagtggcag catatagaac
     2701 agctaaaggg tagtgctgaa ggaagcatac gataccccgc atggaatggg ataatatcac
     2761 aggaggtact agactacctt tcatcctaca taaatagacg catataagag tgaaattctg
     2821 gaaatctgga aatctggttt tgtattcttg ttattcttct ttttgttatt acatatataa
     2881 cttgttactt ttttaaaaaa atctttgtat attttataaa tatataaaac taaatttaag
     2941 aaaaagagaa aaatgtttta tttgagagat tgaaatttta cttgaattta gcttagcttt
     3001 tataaagtat tattatgtaa aaaaacaaaa caaatataca ttaaaaagtt aagactataa
     3061 aatagccacc caaggcattt ctatatcttg ttgttgttgt tttcatcttc tgtatcagag
     3121 gaacttattt tattattttc gtcacgggta ttttctcttg tttgatgatt catcccattc
     3181 attccatcat aaaatgtcga gcgtcaaaac tagagaataa taaagaaaac gatcttttca
     3241 aaaagaaaaa accttttagt tttcctttgt tgttgttgtg ggtgtgtgct atttatatta
     3301 tatagtttac tcataatacc ataaaatatt cggtttgatt aggttatttt aataagctaa
     3361 tttgtttcta atcgtgtaat ttatgctgtg tatattaagt agtgtgtgca ctgcccaaaa
     3421 atgtttgttg tttatagtcg gttaaagaga aaaaagaaaa aaagatccat acacacacgt
     3481 taattagttg ttcaacgtaa tacactcata ttttgttctt atttgctttc ggtcgctgtt
     3541 ctcaccaaga tttattgcca acgaaacaat ttttttttat atattttcag atttttcttt
     3601 ttttcctttc ctttcctttt ctaattttca ctcctggttt tctttctttc ttagaaacat
     3661 tatctcgata ttaatattaa aaaaatataa tcattcaaaa tggacggtgg tatgttttag
     3721 tttagcttca attctaattg attgattaat cagttgattg gtttcaatat gacaaatggg
     3781 tagggtggga aaacttcatt ttcaattcag atcaaacttt tttgttgtcg acataatatt
     3841 tctcgtttgg gatgttactg tcacattaat aatacacaca catcagctta taattttgaa
     3901 agtaatttat cagatatgtt gtgacgatca gtggaaatgg ctaacttcaa tgtatctgtt
     3961 cttccccttt ttcaaagttc acgttttttg attgattgat tgatctgtcg gcagtggttt
     4021 caaaaccatt cggtgagtaa tcctatcaat caatgttacg acaaaaggct caatattcaa
     4081 aattgcaatg ttttatgttt tcctacgtgt acttgtgcaa ggcaattgat tcaacattgc
     4141 ttttggtgtt tgacgagttt ctagtttgga cttgtgttgt tatctgggct atacagattt
     4201 cccggctcac tatgaatttt ttttttcgac gctcagtgca cacaactata aacaacacaa
     4261 acacaaacac agcaagaaaa aaaaaaaacg aacattgaat tgaaaccaag ccaactgaaa
     4321 aattccttat ttaaatgact gtcatactaa cccattttta tagaagaagt tgctgcttta
     4381 gttatcgata acggttctca tatgaaaatt tcggtgatcc ctgagcaggt ggcggaaaca
     4441 ttggatgctg agaaccattt cattgttcgt gaagtgttcg atgtgcacct atccgaccaa
     4501 ggctttgaac tatctaccag aagtgtgagc ccctaccgga aggattacat ctcggatgat
     4561 gactctgatg aagactctgc ttgctatggc gcattcatcg accaagagct tgtcgggaag
     4621 attgaactca actcaacatg gaacgatcta gcctctatcg aacacattgt tgtgtcgcac
     4681 acgcaccgag gcaaaggagt cgcgcacagt ctcatcgaat ttgcgaaaaa gtgggcacta
     4741 agcagacagc tccttggcat acgattagag acacaaacga acaatgtacc tgcctgcaat
     4801 ttgtacgcaa aatgtggctt tactctcggc ggcattgacc tcttcacgta taaaactaga
     4861 cctcaagtct cgaacgaaac agcgatgtac tggtactggt tctcgggagc acaggatgac
     4921 gcctaacata tgtgaagtgt gaagggggag attttcactt tattagattt gtatatatgt
     4981 ataataaata aataaataag ttaaataaat aattagataa gggtggtaat tattactatt
     5041 tacaatcaaa ggtggtcctg caggaagttc ctatactttc tagagaatag gaacttcaga
     5101 tccactagtt ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg
     5161 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5221 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5281 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5341 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5401 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5461 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5521 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5581 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5641 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5701 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5761 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5821 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5881 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5941 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6001 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6061 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6121 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6181 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6241 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tcgaccgaat
     6301 aaatacctgt gacggaagat cacttcgcag aataaataaa tcctggtgtc cctgttgata
     6361 ccgggaagcc ctgggccaac ttttggcgaa aatgagacgt tgatcggcac gtaagaggtt
     6421 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttgtcgagat
     6481 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     6541 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     6601 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     6661 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     6721 tacgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     6781 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     6841 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     6901 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     6961 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     7021 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     7081 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     7141 atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg
     7201 gttgctacgc ctgaataagt gataataagc ggatgaatgg cagaaattcg aaagcaaatt
     7261 cgacccggtc gtcggttcag ggcagggtcg ttaaatagcc gcttatgtct attgctggtt
     7321 taccggttta ttgactaccg gaagcagtgt gaccgtgtgc ttctcaaatg cctgaggcca
     7381 gtttgctcag gctctccccg tggaggtaat aattgacgat atgatccttt ttttctgatc
     7441 aaaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7501 tgttgagatc
//
