LOCUS       62056_19615        8238 bp DNA     circular SYN 14-APR-2019
DEFINITION  Cloning vector pSFS2A-mNeonGreen, complete sequence.
ACCESSION   MK431400
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8238)
  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 8238)
  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 8238)
  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"
FEATURES             Location/Qualifiers
     source          1..8238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    30..41
                     /label=GGSG linker
                     /note="GGSG linker"
     CDS             42..749
                     /label=mNeonGreen
                     /note="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
     misc_recomb     complement(759..792)
                     /label=FLP recombination target sequence (FRT)
                     /note="FLP recombination target sequence (FRT)"
     misc_feature    797..1344
                     /label=promoter region from Candida albicans MAL2
                     /note="promoter region from Candida albicans MAL2"
     CDS             1351..2619
                     /label=FLP
                     /note="site-specific recombinase"
     misc_feature    2623..3010
                     /label=terminator region from Candida albicans ACT1
                     /note="terminator region from Candida albicans ACT1"
     misc_feature    3017..3514
                     /label=promoter region from Candida albicans ACT1
                     /note="promoter region from Candida albicans ACT1"
     gene            3515..4741
                     /gene="SAT1"
                     /label=SAT1
     CDS             join(3515..3524,4179..4741)
                     /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    4745..4874
                     /label=terminator region from Candida albicans URA3
                     /note="terminator region from Candida albicans URA3"
     protein_bind    complement(4879..4912)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     primer_bind     complement(4914..4930)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4967..4985)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5006..5022)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5030..5046)
                     /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(5054..5084)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5099..5120)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5408..5996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6216..6318
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             6319..6975
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(7466..7571)
                     /label=AmpR promoter
     rep_origin      7598..8053
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     8194..8210
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8220..8238
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcgaattggg taccgggccc cccctcgagg gtggtagtgg tatggtttct aaaggtgaag
       61 aagataatat ggctagtttg cctgcaacac atgaattaca tatttttggt agtattaatg
      121 gtgttgattt tgatatggtt ggtcaaggta ctggtaatcc aaatgatggt tatgaagaat
      181 tgaatttgaa atcaactaaa ggagatttgc aattttctcc atggattttg gttccacata
      241 ttggttatgg tttccatcaa tatttgccat atccagatgg aatgtcacca tttcaagctg
      301 ctatggttga tggttctggt tatcaagttc atagaactat gcaattcgaa gatggtgctt
      361 cattgactgt taattataga tatacttatg aaggttctca tattaaaggt gaagctcaag
      421 ttaaaggtac tggttttcca gctgatggtc cagttatgac taattcatta actgctgctg
      481 attggtgtag atcaaagaaa acttatccaa atgataaaac tattatttca acttttaaat
      541 ggtcttatac tactggtaat ggtaaaagat atagatcaac tgctagaact acttatactt
      601 tcgctaaacc aatggctgct aattatttga aaaatcaacc aatgtatgtt tttagaaaaa
      661 ctgaattaaa acattctaaa actgaattga atttcaaaga atggcaaaaa gcattcactg
      721 atgttatggg tatggatgaa ttgtataaat agctcgagga agttcctata ctttctagag
      781 aataggaact tcggatccaa taatgattgg tttgatattt ttgtctagta ccatctgtac
      841 cattacactt aaattatctt tatatctgtc taactcgact gtctggattt cattgatgta
      901 gtcgtatgca tcgttagttc caaaaaatat tgtcatcaat ttgatattgg tttccgactc
      961 taaaattttt ggaagaattt gtctagcgtg ctctgagttg tagccactga aaccacggtt
     1021 aataacatcc aattttcgga tatacacatt ctgtaatgct ggatgaaagc catactgggt
     1081 acaactaaac tgggtgatgg agtcaccgaa caacacaaat ttaccgtatt ccatgattgc
     1141 tatggttgag aatttttttt ttttcttgtc ccacgccatt tttcaaatta tgcagttgag
     1201 aatgttagtt tttgtgtaca ccccgttcgc tgaatatttc ggaataattc aaagattggg
     1261 gagtggggga ggcgatagac gaagacacgg tataaaaatg ggcaaaattt tccccaactt
     1321 tttgcagtgg tttaactaat aatcgtcgac atgccacaat ttgatatatt atgtaaaaca
     1381 ccacctaagg tgcttgttcg tcagtttgtg gaaaggtttg aaagaccttc aggtgagaaa
     1441 atagcattat gtgctgctga actaacctat ttatgttgga tgattacaca taacggaaca
     1501 gcaatcaaga gagccacatt catgagctat aatactatca taagcaattc gttgagtttc
     1561 gatattgtca ataaatcact ccagtttaaa tacaagacgc aaaaagcaac aattttggaa
     1621 gcctcattaa agaaattgat tcctgcttgg gaatttacaa ttattcctta ctatggacaa
     1681 aaacatcaat ctgatatcac tgatattgta agtagtttgc aattacagtt cgaatcatcg
     1741 gaagaagcag ataagggaaa tagccacagt aaaaaaatgc ttaaagcact tctaagtgag
     1801 ggtgaaagca tctgggagat cactgagaaa atactaaatt cgtttgagta tacttcgaga
     1861 tttacaaaaa caaaaacttt ataccaattc ctcttcctag ctactttcat caattgtgga
     1921 agattcagcg atattaagaa cgttgatccg aaatcattta aattagtcca aaataagtat
     1981 ttgggagtaa taatccagtg tttagtgaca gagacaaaga caagcgttag taggcacata
     2041 tacttcttta gcgcaagggg taggatcgat ccacttgtat atttggatga atttttgagg
     2101 aattctgaac cagtcctaaa acgagtaaat aggaccggca attcttcaag caataaacag
     2161 gaataccaat tattaaaaga taacttagtc agatcgtaca ataaagcttt gaagaaaaat
     2221 gcgccttatt caatctttgc tataaaaaat ggcccaaaat ctcacattgg aagacatttg
     2281 atgacctcat ttctttcaat gaagggccta acggagttga ctaatgttgt gggaaattgg
     2341 agcgataagc gtgcttctgc cgtggccagg acaacgtata ctcatcagat aacagcaata
     2401 cctgatcact acttcgcact agtttctcgg tactatgcat atgatccaat atcaaaggaa
     2461 atgatagcat tgaaggatga gactaatcca attgaggagt ggcagcatat agaacagcta
     2521 aagggtagtg ctgaaggaag catacgatac cccgcatgga atgggataat atcacaggag
     2581 gtactagact acctttcatc ctacataaat agacgcatat aagagtgaaa ttctggaaat
     2641 ctggaaatct ggttttgtat tcttgttatt cttctttttg ttattacata tataacttgt
     2701 tactttttta aaaaaatctt tgtatatttt ataaatatat aaaactaaat ttaagaaaaa
     2761 gagaaaaatg ttttatttga gagattgaaa ttttacttga atttagctta gcttttataa
     2821 agtattatta tgtaaaaaaa caaaacaaat atacattaaa aagttaagac tataaaatag
     2881 ccacccaagg catttctata tcttgttgtt gttgttttca tcttctgtat cagaggaact
     2941 tattttatta ttttcgtcac gggtattttc tcttgtttga tgattcatcc cattcattcc
     3001 atcataaaat gtcgagcgtc aaaactagag aataataaag aaaacgatct tttcaaaaag
     3061 aaaaaacctt ttagttttcc tttgttgttg ttgtgggtgt gtgctattta tattatatag
     3121 tttactcata ataccataaa atattcggtt tgattaggtt attttaataa gctaatttgt
     3181 ttctaatcgt gtaatttatg ctgtgtatat taagtagtgt gtgcactgcc caaaaatgtt
     3241 tgttgtttat agtcggttaa agagaaaaaa gaaaaaaaga tccatacaca cacgttaatt
     3301 agttgttcaa cgtaatacac tcatattttg ttcttatttg ctttcggtcg ctgttctcac
     3361 caagatttat tgccaacgaa acaatttttt tttatatatt ttcagatttt tctttttttc
     3421 ctttcctttc cttttctaat tttcactcct ggttttcttt ctttcttaga aacattatct
     3481 cgatattaat attaaaaaaa tataatcatt caaaatggac ggtggtatgt tttagtttag
     3541 cttcaattct aattgattga ttaatcagtt gattggtttc aatatgacaa atgggtaggg
     3601 tgggaaaact tcattttcaa ttcagatcaa acttttttgt tgtcgacata atatttctcg
     3661 tttgggatgt tactgtcaca ttaataatac acacacatca gcttataatt ttgaaagtaa
     3721 tttatcagat atgttgtgac gatcaatgga aatggctaac ttcaatgtat ctgttcttcc
     3781 cctttttcaa agttcacgtt ttttgattga ttgattgatc tgtcggcagt ggtttcaaaa
     3841 ccattcggtg agtaatccta tcaatcaatg ttacgacaaa aggctcaata ttcaaaattg
     3901 caatgtttta tgttttccta cgtgtacttg tgcaaggcaa ttgattcaac attgcttttg
     3961 gtgtttgacg agtttctagt ttggacttgt gttgttatct gggctataca gatttcccgg
     4021 ctcactatga attttttttt tcgacgctca gtgcacacaa ctataaacaa cacaaacaca
     4081 aacacagcaa gaaaaaaaaa aaacgaacat tgaattgaaa ccaagccaac tgaaaaattc
     4141 cttatttaaa tgactgtcat actaacccat ttttatagaa gaagttgctg ctttagttat
     4201 cgataacggt tctcatatga aaatttcggt gatccctgag caggtggcgg aaacattgga
     4261 tgctgagaac catttcattg ttcgtgaagt gttcgatgtg cacctatccg accaaggctt
     4321 tgaactatct accagaagtg tgagccccta ccggaaggat tacatctcgg atgatgactc
     4381 tgatgaagac tctgcttgct atggcgcatt catcgaccaa gagcttgtcg ggaagattga
     4441 actcaactca acatggaacg atctagcctc tatcgaacac attgttgtgt cgcacacgca
     4501 ccgaggcaaa ggagtcgcgc acagtctcat cgaatttgcg aaaaagtggg cactaagcag
     4561 acagctcctt ggcatacgat tagagacaca aacgaacaat gtacctgcct gcaatttgta
     4621 cgcaaaatgt ggctttactc tcggcggcat tgacctcttc acgtataaaa ctagacctca
     4681 agtctcgaac gaaacagcga tgtactggta ctggttctcg ggagcacagg atgacgccta
     4741 acatatgtga agtgtgaagg gggagatttt cactttatta gatttgtata tatgtataat
     4801 aaataaataa ataagttaaa taaataatta gataagggtg gtaattatta ctatttacaa
     4861 tcaaaggtgg tcctgcagga agttcctata ctttctagag aataggaact tcagatccac
     4921 tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt
     4981 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5041 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5101 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5161 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5221 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5281 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5341 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5401 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5461 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5521 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5581 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5641 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5701 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5761 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5821 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     5881 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5941 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6001 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6061 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttcgac cgaataaata
     6121 cctgtgacgg aagatcactt cgcagaataa ataaatcctg gtgtccctgt tgataccggg
     6181 aagccctggg ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac
     6241 tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttgtc gagattttca
     6301 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     6361 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     6421 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     6481 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattacgt
     6541 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     6601 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     6661 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     6721 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
     6781 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     6841 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     6901 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     6961 tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggttgc
     7021 tacgcctgaa taagtgataa taagcggatg aatggcagaa attcgaaagc aaattcgacc
     7081 cggtcgtcgg ttcagggcag ggtcgttaaa tagccgctta tgtctattgc tggtttaccg
     7141 gtttattgac taccggaagc agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg
     7201 ctcaggctct ccccgtggag gtaataattg acgatatgat cctttttttc tgatcaaaag
     7261 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7321 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7381 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7441 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7501 aagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7561 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta
     7621 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     7681 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     7741 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     7801 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     7861 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     7921 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     7981 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     8041 acgcttacaa tttccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     8101 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     8161 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt
     8221 aatacgactc actatagg
//
