LOCUS       pSUC2-yEGFPCLN2P        6515 bp    DNA     circular UNA 04-MAR-2015
DEFINITION  Digestion of pSUC2 - Fragment 2 inserted into yEGFP3-CLN2PEST-
            pRS406.
ACCESSION   urn.local...8h-46io5ta
VERSION     urn.local...8h-46io5ta
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     source          1..>2023
                     /organism="pRS406"
                     /mol_type="other DNA"
     misc_feature    complement(29..51)
                     /label="pGEX_3_primer"
     promoter        202..415
                     /label="URA3_promoter"
     gene            417..1217
                     /gene="URA3"
                     /label="URA3"
     CDS             417..1220
                     /gene="URA3"
                     /note="ORF frame 3"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKEL
                     LELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVK
                     LQYSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLS
                     TGEYTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYR
                     TVDDVVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN*"
                     /label="URA3"
     rep_origin      complement(1483..1789)
                     /label="f1_origin"
     misc_feature    complement(1822..1965)
                     /label="lacZ_a"
     misc_feature    1936..1958
                     /label="M13_pUC_fwd_primer"
     misc_feature    1951..1967
                     /label="M13_forward20_primer"
     promoter        1977..1995
                     /label="T7_promoter"
     misc_feature    <2020..2024
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     ligation        2020..2023
                     /label="Ligation"
     promoter        2024..2725
                     /created_by="User"
                     /label="SUC2 promoter"
     misc_feature    2725..>2729
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    <2726..2730
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     ligation        2726..2729
                     /label="Ligation"
     misc_feature    2730..4201
                     /label="pFA6a-yEGFP3-CLN2PEST-natMX6 (modified)
                     extraction5"
                     /note="Geneious type: Inserted sequence"
     primer_bind     2731..2756
                     /created_by="primer3"
                     /%GC="23.08"
                     /Tm="47.3"
                     /Hairpin="5.0"
                     /Primer-Dimer="2.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Product_Size="1470.0"
                     /Pair_Hairpin="4.0"
                     /Pair_Primer-Dimer="0.0"
                     /Pair_Tm_Diff="6.85"
                     /Sequence="ATTAAATGTCTAAAGGTGAAGAATTA"
                     /annotation_group="PrimerPlugin
                     Group9_urn:local:.:1314758220459.12"
                     /label="1st forward primer"
     primer_bind     2731..2754
                     /created_by="primer3"
                     /%GC="25.0"
                     /Tm="46.47"
                     /Hairpin="5.0"
                     /Primer-Dimer="2.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Product_Size="1469.0"
                     /Pair_Hairpin="4.0"
                     /Pair_Primer-Dimer="0.0"
                     /Pair_Tm_Diff="6.53"
                     /Sequence="ATTAAATGTCTAAAGGTGAAGAAT"
                     /annotation_group="PrimerPlugin
                     Group5_urn:local:.:1314758220459.12"
                     /label="5th forward primer"
     primer_bind     2731..2753
                     /created_by="primer3"
                     /%GC="26.09"
                     /Tm="45.86"
                     /Hairpin="5.0"
                     /Primer-Dimer="0.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Product_Size="1470.0"
                     /Pair_Hairpin="4.0"
                     /Pair_Primer-Dimer="1.0"
                     /Pair_Tm_Diff="6.84"
                     /Sequence="ATTAAATGTCTAAAGGTGAAGAA"
                     /annotation_group="PrimerPlugin
                     Group8_urn:local:.:1314758220459.12"
                     /label="2nd forward primer"
     primer_bind     2732..2754
                     /created_by="primer3"
                     /%GC="26.09"
                     /Tm="45.86"
                     /Hairpin="4.0"
                     /Primer-Dimer="2.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Product_Size="1469.0"
                     /Pair_Hairpin="4.0"
                     /Pair_Primer-Dimer="0.0"
                     /Pair_Tm_Diff="6.84"
                     /Sequence="TTAAATGTCTAAAGGTGAAGAAT"
                     /annotation_group="PrimerPlugin
                     Group7_urn:local:.:1314758220459.12"
                     /label="3rd forward primer"
     primer_bind     2734..2756
                     /created_by="primer3"
                     /%GC="26.09"
                     /Tm="45.86"
                     /Hairpin="4.0"
                     /Primer-Dimer="2.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Product_Size="1467.0"
                     /Pair_Hairpin="4.0"
                     /Pair_Primer-Dimer="1.0"
                     /Pair_Tm_Diff="6.84"
                     /Sequence="AAATGTCTAAAGGTGAAGAATTA"
                     /annotation_group="PrimerPlugin
                     Group6_urn:local:.:1314758220459.12"
                     /label="4th forward primer"
     ORF             2736..3452
                     /created_by="User"
                     /label="EGFP"
     CDS             2739..3986
                     /gene="yEGFP3-CLN2Pest"
                     /product="modified unstable green fluorescent protein"
     misc_feature    3994..4196
                     /gene="ADH1t"
                     /product="ADH1 terminator"
     misc_feature    complement(4181..4200)
                     /created_by="primer3"
                     /%GC="60.0"
                     /Tm="54.15"
                     /Hairpin="8.0"
                     /Primer-Dimer="0.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Product_Size="1470.0"
                     /Pair_Hairpin="4.0"
                     /Pair_Primer-Dimer="0.0"
                     /Pair_Tm_Diff="6.85"
                     /Sequence="GATCTGCCGGTAGAGGTGTG"
                     /annotation_group="PrimerPlugin
                     Group9_urn:local:.:1314758220459.12"
                     /label="1st reverse primer"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(4181..4199)
                     /created_by="primer3"
                     /%GC="57.9"
                     /Tm="53.0"
                     /Hairpin="8.0"
                     /Primer-Dimer="0.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Product_Size="1469.0"
                     /Pair_Hairpin="4.0"
                     /Pair_Primer-Dimer="0.0"
                     /Pair_Tm_Diff="6.53"
                     /Sequence="ATCTGCCGGTAGAGGTGTG"
                     /annotation_group="PrimerPlugin
                     Group5_urn:local:.:1314758220459.12"
                     /label="5th reverse primer"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(4182..4200)
                     /created_by="primer3"
                     /%GC="57.9"
                     /Tm="52.7"
                     /Hairpin="8.0"
                     /Primer-Dimer="0.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Product_Size="1469.0"
                     /Pair_Hairpin="4.0"
                     /Pair_Primer-Dimer="0.0"
                     /Pair_Tm_Diff="6.84"
                     /Sequence="GATCTGCCGGTAGAGGTGT"
                     /annotation_group="PrimerPlugin
                     Group7_urn:local:.:1314758220459.12"
                     /label="3rd reverse primer"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(4182..4200)
                     /created_by="primer3"
                     /%GC="57.9"
                     /Tm="52.7"
                     /Hairpin="8.0"
                     /Primer-Dimer="0.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Product_Size="1467.0"
                     /Pair_Hairpin="4.0"
                     /Pair_Primer-Dimer="1.0"
                     /Pair_Tm_Diff="6.84"
                     /Sequence="GATCTGCCGGTAGAGGTGT"
                     /annotation_group="PrimerPlugin
                     Group6_urn:local:.:1314758220459.12"
                     /label="4th reverse primer"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(4182..4200)
                     /created_by="primer3"
                     /%GC="57.9"
                     /Tm="52.7"
                     /Hairpin="8.0"
                     /Primer-Dimer="0.0"
                     /Monovalent_Salt_Concentration="50.0"
                     /Divalent_Salt_Concentration="0.0"
                     /DNTP_Concentration="0.0"
                     /Product_Size="1470.0"
                     /Pair_Hairpin="4.0"
                     /Pair_Primer-Dimer="1.0"
                     /Pair_Tm_Diff="6.84"
                     /Sequence="GATCTGCCGGTAGAGGTGT"
                     /annotation_group="PrimerPlugin
                     Group8_urn:local:.:1314758220459.12"
                     /label="2nd reverse primer"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    4201
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     source          <4202..6515
                     /organism="pRS406"
                     /mol_type="other DNA"
     ligation        4202..4205
                     /label="Ligation"
     misc_feature    complement(4202..4218)
                     /label="pBluescriptSK_primer"
     promoter        complement(4254..4273)
                     /label="T3_promoter"
     misc_feature    complement(4290..4308)
                     /label="M13_reverse_primer"
     misc_feature    complement(4307..4329)
                     /label="M13_pUC_rev_primer"
     rep_origin      complement(4681..5300)
                     /label="pBR322_origin"
     gene            complement(5455..6315)
                     /gene="Ampicillin"
                     /label="Ampicillin"
     CDS             complement(5455..6315)
                     /gene="Ampicillin"
                     /note="ORF frame 3"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /label="Ampicillin"
     promoter        complement(6357..6385)
                     /label="AmpR_promoter"
ORIGIN      
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     1981 acgactcact atagggcgaa ttgggtaccg ggccccccct cgagacatac taagacattt
     2041 accgtatggg agttgttgtc ctagcgtagt tctcgctccc ccagcaaagc tcaaaaaagt
     2101 acgtcattta gaatagtttg tgagcaaatt accagtcggt atgctacgtt agaaaggccc
     2161 acagtattct tctaccaaag gcgtgccttt gttgaactcg atccattatg agggcttcca
     2221 ttattccccg catttttatt actctgaaca ggaataaaaa gaaaaaaccc agtttaggaa
     2281 attatccggg ggcgaagaaa tacgcgtagc gttaatcgac cccacgtcca gggtttttcc
     2341 atggaggttt ctggaaaaac tgacgaggaa tgtgattata aatcccttta tgtgatgtct
     2401 aagactttta aggtacgccc gatgtttgcc tattaccatc atagagacgt ttcttttcga
     2461 ggaatgctta aacgactttg tttgacaaaa atgttgccta agggctctat agtaaaccat
     2521 ttggaagaaa gatttgacga cttttttttt ttggatttcg atcctataat ccttcctcct
     2581 gaaaagaaac atataaatag atatgtatta ttcttcaaaa cattctcttg ttcttgtgct
     2641 ttttttttac catatatctt actttttttt ttctctcaga gaaacaagca aaacaaaaag
     2701 cttttctttt cactaacgta tatggaattc attaaatgtc taaaggtgaa gaattattca
     2761 ctggtgttgt cccaattttg gttgaattag atggtgatgt taatggtcac aaattttctg
     2821 tctccggtga aggtgaaggt gatgctactt acggtaaatt gaccttaaaa tttatttgta
     2881 ctactggtaa attgccagtt ccatggccaa ccttagtcac tactttcggt tatggtgttc
     2941 aatgttttgc gagataccca gatcatatga aacaacatga ctttttcaag tctgccatgc
     3001 cagaaggtta tgttcaagaa agaactattt ttttcaaaga tgacggtaac tacaagacca
     3061 gagctgaagt caagtttgaa ggtgatacct tagttaatag aatcgaatta aaaggtattg
     3121 attttaaaga agatggtaac attttaggtc acaaattgga atacaactat aactctcaca
     3181 atgtttacat catggctgac aaacaaaaga atggtatcaa agttaacttc aaaattagac
     3241 acaacattga agatggttct gttcaattag ctgaccatta tcaacaaaat actccaattg
     3301 gtgatggtcc agtcttgtta ccagacaacc attacttatc cactcaatct gccttatcca
     3361 aagatccaaa cgaaaagaga gaccacatgg tcttgttaga atttgttact gctgctggta
     3421 ttacccatgg tatggatgaa ttgtacaaag catccaactt gaacatttcg agaaagctta
     3481 ccatatcaac cccatcatgc tctttcgaaa attcaaatag cacatccatt ccttcgcccg
     3541 cttcctcatc tcaaagccac actccaatga gaaacatgag ctcactctct gataacagcg
     3601 ttttcagccg gaatatggaa caatcatcac caatcactcc aagtatgtac caatttggtc
     3661 agcagcagtc aaacagtata tgtggtagca ccgttagtgt gaatagtctg gtgaatacaa
     3721 ataacaaaca aaggatctac gaacaaatca cgggtcctaa cagcaataac gcaaccaatg
     3781 attatattga tttgctaaac ctaaatgagt ctaacaagga aaaccaaaat cccgcaacgg
     3841 cgcattacct caatgggggc ccacccaaga caagcttcat taaccatgga atgttcccct
     3901 cgccaactgg gaccataaat agcggtaaat ctagcagtgc ctcatcttta atttcttttg
     3961 gtatgggcaa tacccaagta atatagggcg cgccacttct aaataagcga atttcttatg
     4021 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
     4081 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag
     4141 gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta ccggcagatc
     4201 ggatccacta gttctagagc ggccgccacc gcggtggagc tccagctttt gttcccttta
     4261 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4321 ttatccgctc acaattccac acaacatagg agccggaagc ataaagtgta aagcctgggg
     4381 tgcctaatga gtgaggtaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4441 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4501 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4561 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4621 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4681 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4741 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4801 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4861 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4921 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4981 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5041 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5101 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     5161 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5221 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5281 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5341 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5401 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5461 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5521 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5581 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5641 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5701 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5761 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5821 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5881 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5941 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6001 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6061 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6121 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6181 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6241 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6301 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6361 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6421 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6481 ctataaaaat aggcgtatca cgaggccctt tcgtc
//
