LOCUS       Exported                7509 bp DNA     circular SYN 20-JUL-2025
DEFINITION  Cloning vector pSFS2A-mNeonGreen, complete sequence.
ACCESSION   MK431400
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7509)
  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 7509)
  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 7509)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7509)
  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
            (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..7509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(915..7509,1..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..4925
                     /gene="SAT1"
                     /label=SAT1
     CDS             join(3700..3709,4363..4925)
                     /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    4929..5058
                     /label=terminator region from Candida albicans URA3
                     /note="terminator region from Candida albicans URA3"
     protein_bind    complement(5063..5096)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     primer_bind     complement(5098..5114)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5151..5169)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5190..5206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5214..5230)
                     /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(5238..5268)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5283..5304)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5592..6180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6400..6502
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             6503..7159
                     /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 aaaaaaacga acattgaatt gaaaccaagc caactgaaaa
     4321 attccttatt taaatgactg tcatactaac ccatttttat agaagaagtt gctgctttag
     4381 ttatcgataa cggttctcat atgaaaattt cggtgatccc tgagcaggtg gcggaaacat
     4441 tggatgctga gaaccatttc attgttcgtg aagtgttcga tgtgcaccta tccgaccaag
     4501 gctttgaact atctaccaga agtgtgagcc cctaccggaa ggattacatc tcggatgatg
     4561 actctgatga agactctgct tgctatggcg cattcatcga ccaagagctt gtcgggaaga
     4621 ttgaactcaa ctcaacatgg aacgatctag cctctatcga acacattgtt gtgtcgcaca
     4681 cgcaccgagg caaaggagtc gcgcacagtc tcatcgaatt tgcgaaaaag tgggcactaa
     4741 gcagacagct ccttggcata cgattagaga cacaaacgaa caatgtacct gcctgcaatt
     4801 tgtacgcaaa atgtggcttt actctcggcg gcattgacct cttcacgtat aaaactagac
     4861 ctcaagtctc gaacgaaaca gcgatgtact ggtactggtt ctcgggagca caggatgacg
     4921 cctaacatat gtgaagtgtg aagggggaga ttttcacttt attagatttg tatatatgta
     4981 taataaataa ataaataagt taaataaata attagataag ggtggtaatt attactattt
     5041 acaatcaaag gtggtcctgc aggaagttcc tatactttct agagaatagg aacttcagat
     5101 ccactagttc tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga
     5161 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5221 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5281 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5341 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5401 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5461 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5521 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5581 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5641 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5701 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5761 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5821 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5881 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5941 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6001 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6061 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6121 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6181 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6241 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt cgaccgaata
     6301 aatacctgtg acggaagatc acttcgcaga ataaataaat cctggtgtcc ctgttgatac
     6361 cgggaagccc tgggccaact tttggcgaaa atgagacgtt gatcggcacg taagaggttc
     6421 caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt tgtcgagatt
     6481 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     6541 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     6601 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     6661 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     6721 acgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     6781 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     6841 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     6901 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     6961 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     7021 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     7081 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     7141 tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg
     7201 ttgctacgcc tgaataagtg ataataagcg gatgaatggc agaaattcga aagcaaattc
     7261 gacccggtcg tcggttcagg gcagggtcgt taaatagccg cttatgtcta ttgctggttt
     7321 accggtttat tgactaccgg aagcagtgtg accgtgtgct tctcaaatgc ctgaggccag
     7381 tttgctcagg ctctccccgt ggaggtaata attgacgata tgatcctttt tttctgatca
     7441 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7501 gttgagatc
//
