LOCUS       sequence-472645-        9374 bp    DNA     circular SYN 14-MAY-2025
DEFINITION  Expression of FL ScVPS38-3xFLAG under its own promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9374)
  AUTHORS   Rostislavleva K, Soler N, Ohashi Y, Zhang L, Pardon E, Burke JE, 
            Masson GR, Johnson C, Steyaert J, Ktistakis NT, Williams RL
  TITLE     Structure and flexibility of the endosomal Vps34 complex reveals the
            basis of its function on membranes.
  JOURNAL   Science. 2015 Oct 9;350(6257):aac7365. doi: 10.1126/science.aac7365.
  PUBMED    26450213
REFERENCE   2  (bases 1 to 9374)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2015-10-9"; volume: "350(6257):aac7365. doi"; pages: " 
            10.1126/science.aac7365"
COMMENT     Sequence Label: pYO759
FEATURES             Location/Qualifiers
     source          1..9374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..81
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     73..90
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     74..90
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     100..119
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        100..118
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            complement(174..1293)
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     terminator      complement(174..371)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(377..949)
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     promoter        complement(950..1293)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     protein_bind    1381..1498
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     1679..1702
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             1835..1855
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     gap             1924..2023
                     /estimated_length=100
     CDS             2145..2318
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2328..2348
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2379..2399
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2421..2441
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(7183..7203)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7183..7201)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7222..7238)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7222..7238)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7235..7257)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7246..7262
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(join(7270..7276,7277..7294,7295..7300))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7315..7336
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7453..7470)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7624..8212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7704..7723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8383..9174,9175..9243))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9006..9025
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9244..9348)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
       61 cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc
      121 gaattggagc tcgttttcga cactggatgg cggcgttagt atcgaatcga cagcagtata
      181 gcgaccagca ttcacatacg attgacgcat gatattactt tctgcgcact taacttcgca
      241 tctgggcaga tgatgtcgag gcgaaaaaaa atataaatca cgctaacatt tgattaaaat
      301 agaacaacta caatataaaa aaactataca aatgacaagt tcttgaaaac aagaatcttt
      361 ttattgtcag tactgattag gggcagggca tgctcatgta gagcgcctgc tcgccgtccg
      421 aggcggtgcc gtcgtacagg gcggtgtcca ggccgcagag ggtgaacccc atccgccggt
      481 acgcgtggat cgccggtgcg ttgacgttgg tgacctccag ccagaggtgc ccggcgcccc
      541 gctcgcgggc gaactccgtc gcgagcccca tcaacgcgcg cccgaccccg tgcccccggt
      601 gctccggggc gacctcgatg tcctcgacgg tcagccggcg gttccagccg gagtacgaga
      661 cgaccacgaa gcccgccagg tcgccgtcgt ccccgtacgc gacgaacgtc cgggagtccg
      721 ggtcgccgtc ctccccgtcg tccgattcgt cgtccgattc gtcgtcgggg aacaccttgg
      781 tcaggggcgg gtccaccggc acctcccgca gggtgaagcc gtccccggtg gcggtgacgc
      841 ggaagacggt gtcggtggtg aaggacccat ccagtgcctc gatggcctcg gcgtcccccg
      901 ggacactggt gcggtaccgg taagccgtgt cgtcaagagt ggtacccatg gttgtttatg
      961 ttcggatgtg atgtgagaac tgtatcctag caagatttta aaaggaagta tatgaaagaa
     1021 gaacctcagt ggcaaatcct aaccttttat atttctctac aggggcgcgg cgtggggaca
     1081 attcaacgcg tctgtgaggg gagcgtttcc ctgctcgcag gtctgcagcg aggagccgta
     1141 atttttgctt cgcgccgtgc ggccatcaaa atgtatggat gcaaatgatt atacatgggg
     1201 atgtatgggc taaatgtacg ggcgacagtc acatcatgcc cctgagctgc gcacgtcaag
     1261 actgtcaagg agggtattct gggcctccat gtcgctggcc gggtgacccg gcggggacga
     1321 ggcaagctaa acagatctgc ggccgcggat ctttgcaaag atccaaaaaa agcatgcgta
     1381 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc
     1441 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
     1501 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaattg gcagtaacct
     1561 ggccccacaa acctcaaatg aacgaatcaa attaacaacc agatatctcg actgaaaaaa
     1621 aaggtttaaa ccagttccct gaaattattc ccctacttga ctaataagta tataaagacg
     1681 gtaggtattg attgtaattc tgtaaatcta tttcttaaac ttcttgaatt ctacttttat
     1741 agttagtctt ttttttagtt ttaaaacacc aagaacttag tttcgaataa acacacataa
     1801 ataaaccatg atgggagcag gggcgggtgc tagcgagaac ctgtacttcc agggcgccca
     1861 cgatgaagcc gtagacaaca aattcaacaa agaacaacaa aacgcgttct atgagatctt
     1921 acannnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     1981 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnntttacct aacttaaacg
     2041 aagaacaacg aaacgccttc atccaaagtt taaaagatga cccaagccaa agcgctaacc
     2101 ttttagcaga agctaaaaag ctaaatgatg ctcaggcgcc gaaagtagac aacaaattca
     2161 acaaagaaca acaaaacgcg ttctatgaga tcttacattt acctaactta aacgaagaac
     2221 aacgaaacgc cttcatccaa agtttaaaag atgacccaag ccaaagcgct aaccttttag
     2281 cagaagctaa aaagctaaat gatgctcagg cgccgaaagt agacgcggaa aacctgtatt
     2341 tccagggagg cggatcttct agaccaacta ctgcctctga aaatttgtac tttcaaggcg
     2401 agttgaagac cactgcttct gaaaacttgt acttccaagg tgagttactc gagggtgaat
     2461 tgaagactgc tgctttggct caacacgacg aagccatggg cggatccctt agaagaaagg
     2521 gggagtctgt gaagaaaagg ttggtgtcag gtacagcagc ctcattgcaa gaactctcct
     2581 tacagccttc ttctaatggt attgcatcgg ctggaaagag cgaagatgga gaggctggtg
     2641 atggaatgtc tggagaattt ggggcagcat tgactgattc gaatgtggag aacaatgtta
     2701 aggcatttga tttgatcttt acgaagctta aaagtacgaa tcaactggaa cgagtggcag
     2761 cgtcatacga gttgaaatcg tgtctgattt cgctagctcg agaggtttct acagagtact
     2821 tccagcgttt cagcaatgat ataaataata aggttttcga gttgatccat gggaccgatt
     2881 ctaatgagaa gcttggtggt gtgctagctg ttgatacgtt gattgatttc tattcacata
     2941 ctgatgaatt gcccaatcag acgtcgagat tagcgaatta cttgcgtgtg ttaataccgt
     3001 ccaacgacat cgaggtgatg agagctgcgg cgacgacgtt ggggaagtta gcggtgcccg
     3061 gtgggacatt aacgtcagag tttgttgagt tcgaggtgaa aacttgcatt gaatggttga
     3121 cgacttctcc ggagagttcc tcttcgaatt ccaagcaaga gtacagaaag catgcatctt
     3181 tgttgattat cagttcgatc gcgcacaact cgccatattt actatacccg ttcgtgaact
     3241 cgatcttaga taacatatgg agggctctaa gagacacgaa gctagttatc agaaccgatg
     3301 cagcggtcac gctagggaaa tgtttgacaa ttatcaacga tcgtgactct tctttgacta
     3361 aaaaatgggt acaaagactt ttcaatggct gcatatacgg attgcaacta aatacaacgg
     3421 tgtctataca cggtactcta ttggtataca gacaacttgt gtccctagaa gggggatacc
     3481 tgaaggacaa atatgaagaa atttacgaga caacaatgaa gcatagagac aataagaata
     3541 ttattatcag aaaggaaata tatgctatca ttcctgtttt ggcagcattt gatccaaaac
     3601 tcttcactca aaaatatttg gatgcaacca tgatacatta cctaacacta ttgaagaaca
     3661 tgacttctca ccaaattgcg ctatcggata aaggccctat attagtctcc atcggtgata
     3721 ttgcatatca tgtgggttcc gaaattggcc cataccttga tgccattgta gagaatttaa
     3781 gggatggtct agaaacaaag ttcaaaacta ggaaagaatt cgaaaaggac atattttatt
     3841 gtgttagcaa actagcatgt gctgttaggc cattgttagc caaatattta aacagaggat
     3901 tgcttgacgc tctattggca tgtcccttga gcgatcatat gcaagaaact cttctggtta
     3961 tttgcgaaaa aatcccttcg atggaagtca cagttaatga gaagctttta aatatcattt
     4021 gtttggttct ttcaggggag aagttcagac ctccaggatc accaacgcca atgaaatcgc
     4081 tatcgagcga aactgcaagg gcttatagag accaatcttt actcagaaag acaggtgaaa
     4141 ataataatga tactctcgac gcaatgattc tcagaaaagc tttaagaatg ctttcagata
     4201 ttaaaccaaa atattctttg actgaattta ttaggagagt tataatatcc tacatggaac
     4261 atgataatat gcaagtcaga aaactagctg ctctgacttc ttgtgacttg ttcgtaaagg
     4321 attcaatctg caaacaaacc tcattatatg cattaaatgt ggtttcagag gttttgagta
     4381 agctattaac ggtcgcaatt acagatccag tggcggaaat ccgtctagaa atactaaaac
     4441 atttagacag gaactttgat ccacaacttt ctcaaccaga taatacaaaa cttttattta
     4501 tggcgctgaa tgacgaagtt tttgccatcc aaatggaagc aatgaaaatc tgtggtagac
     4561 ttgccttggt gaacccagca tatgtcattc catctttaag gaaaacttta attcaactat
     4621 tgactgagct aaaacactct aaaatgacaa gaaaaaagga agaatgtgca tcattgctct
     4681 gtactttgat cagttccagt agcgacgtta ccaaacctta ccttgagcct gtgattgaga
     4741 ttttgcttcc aaagtctcaa gatccatctt ctgctgtagc atcaacagcg ttaaaagctt
     4801 taggagaaat atcagtcgtt ggtggtgaag atatgctacc tttcttggat gaattaatgc
     4861 cattaatcat cgatacattc caagatcaat ccaattcttt caagagaggg gctgcactaa
     4921 aggctttagg acaactagct gcctcttcag gatacgttat cgatccttta ttagactacc
     4981 ctgaattact cggtgtgctt atcaatattt tgaaatcaga aagttcacag aatattcgca
     5041 gagaaactgt cagattgatc ggtattctcg gggcattaga tccatacaaa catagagaag
     5101 tggaaagaac atctataaca aatattactt ctgagcaaaa tgcgcctcct attgatgtcg
     5161 cattactaat gcaaggtatg tcaccatcta acgaagaata ctacccaaca gttgtcatcg
     5221 gcgttctaat gaaaatattg aaagatcctt ccttatccat acaccattct actgtcattc
     5281 aagcaataat gcatattttc caaacaatgg gtcttcgttg tgtcattttc ttaaagcaga
     5341 ttattcctgg ttttatactt gtgatgcata cttgtccacc atcccttctt gaactttatt
     5401 tccaacaact cagtgtatta atatccattg taaagcaaca tatcagacca catgttaccg
     5461 aaattgttga tgttatttca gaattcttcc ctgtattgaa gttgcagcta acgatcatat
     5521 ctgttattga atcgttatcg agggctttag aaggtgagtt taatcctcat ttacctacta
     5581 ttctttcttt gtttttagat gttttggaga aagatttgaa tgatattttg gaagctcaaa
     5641 aaattgaatg gcatgaaggt caatcaaaca agaaagttgt atctgttaga attttgaaat
     5701 ctttggtggt gttcggtcct catctagaag aatatgtcca tcttgttcta cctaccatta
     5761 ttaaactatc tgaattcagc tcagggaatt tgaaaaaagc tgcaattatc actattggta
     5821 ggttatccaa gaatgttaat gcattggaaa tgtcttcaag aatcgttcaa gctttggtca
     5881 gagtcttaaa tacttcggaa acggaatacg tgaaagcaac aatgaacacc ttaagtttac
     5941 tacttttgca attaaacatt gatttcactg tattcatacc agtaatcaat aaaacgttgg
     6001 taaaacagaa tatccaacat accatatatg atcgcttagt tgcaaagctt ctaaacaatg
     6061 aacctttacc aacaaaaatt attattgaca aggactttga tttacctaac aaagaaatgg
     6121 atgatgtcaa ggttgcatcg aagaaactcc cggtcaatca aatggttttg aaaaatgctt
     6181 gggattgtag ccaacagaga acaaaagaag attggcaaga gtggattaga agactttcca
     6241 tatctttact caaggaatca tcatctcatg cattgcgtgc atgctctgga ttggcaggta
     6301 tatattaccc attggctaga gaacttttca acgcttcgtt tgcaagttgt tggggtgaac
     6361 tttacacaca gtaccaagag gatttgatac aatcactttg cgcagcttta tcttcaccaa
     6421 acaacccacc tgaaatccat cagactctat taaatttagt tgagttcatg gaacacgatg
     6481 acaaacccct accaatatcg atttcgactt taggcgaata cgcccaacgt tgccatgcct
     6541 atgcaaaggc tcttcattac aaggaattag agtttatcga agaaccttcg acatctacga
     6601 ttgagtcatt aataacaatt aataatcaat tacatcaaac ggatgcggca atcggtatat
     6661 tgaagcacgc tcaacaacac catgatttgc aattgaaaga aacttggtac gagaaacttc
     6721 aaagatggga tgatgcattg aatgcataca acaaacgtga agctgctggg gaagattcag
     6781 tagaggtgac cattggtaaa atgagatctc tacatgcttt gggtgaatgg gatcaactgt
     6841 ctgaactcgc cgctgataag tgggcttctt ctcaactaga agttaagaga gctatcgcac
     6901 cacttgccgc tggtgcagca tggggcttag cacaatggga tagagtagag caatatattg
     6961 aagtgatgaa gccacaatct ccagacaaag ctttctttga cgctattcta tgtctccatc
     7021 gtaacaattt tgaaaaggcg gcggaacaca ttttctctgc cagagatctt ctagttactg
     7081 aaatgtctgc attggtaaat gagagttata atagagctta cggtgttgtg gtaagaactc
     7141 aaatggttgc tgagttgggg gcccggtacc cagcttttgt tccctttagt gagggttaat
     7201 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     7261 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     7321 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     7381 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     7441 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     7501 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7561 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7621 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7681 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7741 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7801 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7861 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7921 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7981 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     8041 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     8101 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     8161 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     8221 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     8281 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     8341 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     8401 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     8461 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     8521 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8581 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8641 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8701 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8761 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8821 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8881 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8941 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     9001 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     9061 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     9121 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     9181 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     9241 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     9301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     9361 aaaagtgcca cctg
//
