LOCUS       Exported                6955 bp DNA     circular SYN 14-NOV-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6955)
  AUTHORS   Jason Yu
  TITLE     Direct Submission
  JOURNAL   Exported Sep 8, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6955
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     165..183
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(221..243)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     325..349
                     /label=HR-MET17-fwd
     CDS             complement(376..1710)
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     promoter        1711..1945
                     /label=MET17 promoter
     primer_bind     complement(1920..1943)
                     /label=HR-MET17_rev
     primer_bind     complement(2140..2161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2352..2371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2588..2610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2602..2619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2603..2619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2626..2645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2626..2644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2647..2670
                     /label=p16NotI_GAP_f
     misc_feature    2653..2734
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     complement(2713..2765)
                     /label=GAPprom_p16_r
     primer_bind     2713..2765
                     /label=p16_GAPprom_f
     primer_bind     complement(2713..2739)
                     /label=uni_p16_r
     primer_bind     2739..2765
                     /label=p16NotI_GAP_f
     promoter        2740..3406
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase"
     primer_bind     2740..2761
                     /label=NotI_GAPprom_f
     primer_bind     complement(3382..3412)
                     /label=mCherry_GAPprom_r
     gene            3431..4147
                     /product="Yeast codon optimised (Pombe) enhanced cyan 
                     fluorescent protein (mEx: 435 nm, mEm: 485 nm). From Watson
                     et al, Gene 2008). Complete sequence homology to pAW8-yECFP
                     (http://www.euroscarf.de/plasmid_details.php?accno=P30547) 
                     "
                     /label=eCFP
     primer_bind     4160..4188
                     /label=ADH1term_mCherry_r
     terminator      4168..4355
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4331..4355)
                     /label=p16SalI_ADH1_r
     primer_bind     complement(4339..4384)
                     /label=p16_ADH1term_r
     primer_bind     complement(4342..4355)
                     /label=SalI_ADH1term_r
     primer_bind     4358..4384
                     /label=ADH1term_p16_f
     primer_bind     4358..4384
                     /label=uni_p16_f
     misc_feature    4358..4362
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(4374..4392)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4374..4392)
                     /label=T3
                     /note="T3 promoter, forward primer"
     primer_bind     complement(4413..4429)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4413..4429)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4426..4448)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4437..4453
                     /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(4461..4491)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4506..4527
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4644..4661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4815..5403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4895..4914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5574..6434)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6197..6216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6435..6539)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(6569..6955,1..117)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac tttctatttt
       61 taatttatat atttatatta aaaaatttaa attataatta tttttatagc acgtgatgaa
      121 aaggacccta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
      181 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
      241 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
      301 cgtcagcggg tgttggcggt cgagcttgtg agagaaagta ggtttataca taattttaca
      361 actcattacg cacactcatg gtttttggcc agcgaaaaca gtttcaaaag attgctggaa
      421 gtctgcaata atgtcatcaa taaattcgat accaacagag acacgaatta agtccttggt
      481 aacaccagat gccaactttt ctttgtcatt taattgtttg tgggtagtga agtatggagc
      541 aatgactaag gtcttggcat caccaacatt ggccaagtta gaggcaagct ttaaattgtc
      601 aacaacttga gcaccagaaa gtttgaatgg gtcagtttcc ttgtcggcat ttggtaagtc
      661 ttttacaccg aaagataaga caccaccgaa accgttagat agatacttct tagcattttc
      721 atgatgagaa tgagatgcta aaccagggta tgaaacccaa gatacgtatg gggattgttc
      781 taaccatttg gctaacttca atgcattttc accgtgtctt tcagctctca aagataatgt
      841 ttcaacacct tgtagtagca agaaagaggc aaatgggttc atcaatggac ccaaatctct
      901 taatagttca gttctaacat gaacgatgta tgccaagtta ccgtaggctt cattgtagat
      961 agtaccgtga tatccttcgg caggttgaga gaattgaggg aacttttctg ggtagtcctt
     1021 ccatgggaac ttaccagagt caacaataat accaccgata gtagtaccat gaccaccaat
     1081 ccatttggta gcagaatgtg ttacaatatc agcaccgtat ttaattggct gacagaagta
     1141 accaccggca ccaaatgtgt tgtcaacgac aactggaata ccgtgtttgt gagcaattgc
     1201 aacaattttt tcaaaatccg gaacattgta ctttggatta ccaatggttt ccaaataaac
     1261 agccttggtt ctttcatcaa agaccttttc gaattcttct ggattgtcac cttcaacaaa
     1321 tctagcctcg ataccaaatc ttttgaacga gattttgaac tggttataag taccaccgta
     1381 taagtaagaa gtggaaacga tgttgtcacc agtgtgtgcc aaaccttgga tggcaagggt
     1441 ttgagcggct tgaccggagg aaacagccaa agcagcagca ccaccttcta aagcagcaat
     1501 tctttcttcc aaaacattac tggttgggtt ttggaaacgg gaatagacgt aacctggaac
     1561 ttctagacca aacaattgcg aaccatgctt agagttttcg aaaacataag aagtggtggc
     1621 gtaaattggt acagctctgg atctgtgagc attgtcacca gggttctctt ggccggcgtg
     1681 tagttgaaca gtatcgaaat gagatggcat tgtatggatg ggggtaatag aattgtatct
     1741 atgtatctga cgaccctgta ttacacgaag gaaatagtag atgaaaagac aagagagcaa
     1801 gaaaaaggaa aaacttcctt tctaacagac gctttactta accttatata tatcttattt
     1861 ttttcatcga gcgtgttcaa ttggacaagg tgccaccttt tcgacacttc ggttattatg
     1921 ttacacagtt ttcatgagga tggcggatcc tatgcggtgt gaaataccgc acagatgcgt
     1981 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     2041 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2101 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2161 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2221 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2281 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2341 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc tggcaagtgt
     2401 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2461 gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     2521 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     2581 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     2641 tagggcgaat tggagctcca ccgcggtggc ggccgctcta gaactagtgg gtcccccggg
     2701 ctgcaggaat tcgatatcaa gcttatcgat accgtcgact cattatcaat actgccattt
     2761 caaagaatac gtaaataatt aatagtagtg attttcctaa ctttatttag tcaaaaaatt
     2821 agccttttaa ttctgctgta acccgtacat gcccaaaata gggggcgggt tacacagaat
     2881 atataacatc gtaggtgtct gggtgaacag tttattcctg gcatccacta aatataatgg
     2941 agcccgcttt ttaagctggc atccagaaaa aaaaagaatc ccagcaccaa aatattgttt
     3001 tcttcaccaa ccatcagttc ataggtccat tctcttagcg caactacaga gaacaggggc
     3061 acaaacaggc aaaaaacggg cacaacctca atggagtgat gcaacctgcc tggagtaaat
     3121 gatgacacaa ggcaattgac ccacgcatgt atctatctca ttttcttaca ccttctatta
     3181 ccttctgctc tctctgattt ggaaaaagct gaaaaaaaag gttgaaacca gttccctgaa
     3241 attattcccc tacttgacta ataagtatat aaagacggta ggtattgatt gtaattctgt
     3301 aaatctattt cttaaacttc ttaaattcta cttttatagt tagtcttttt tttagtttta
     3361 aaacaccaag aacttagttt cgaataaaca cacataaaca aacaaaggtg acggtgctgg
     3421 tttaattaac atgtctaaag gtgaagaatt attcactggt gttgtcccaa ttttggttga
     3481 attagatggt gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc
     3541 tacttacggt aaattgacct taaaatttat ttgtactact ggtaaattgc cagttccatg
     3601 gccaacctta gtcactactt taacttgggg tgttcaatgt ttttctagat acccagatca
     3661 tatgaaacaa catgactttt tcaagtctgc catgccagaa ggttatgttc aagaaagaac
     3721 tatttttttc aaagatgacg gtaactacaa gaccagagct gaagtcaagt ttgaaggtga
     3781 taccttagtt aatagaatcg aattaaaagg tattgatttt aaagaagatg gtaacatttt
     3841 aggtcacaaa ttggaataca tttataactc tcacaatgtt tacatcactg ctgacaaaca
     3901 aaagaatggt atcaaagcta acttcaaaat tagacacaac attgaagatg gttctgttca
     3961 attagctgac cattatcaac aaaatactcc aattggtgat ggtccagtct tgttaccaga
     4021 caaccattac ttatccactc aatctgcctt atccaaagat ccaaacgaaa agagagacca
     4081 catggtcttg ttagaatttg ttactgctgc tggtattacc catggtatgg atgaattgta
     4141 caaataaggc gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt
     4201 aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa
     4261 cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag
     4321 tatgaggtcg ctcttattga ccacacctct accggcagta ccagcttttg ttccctttag
     4381 tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4441 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4501 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4561 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4621 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4681 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4741 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4801 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4861 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4921 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4981 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5041 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5101 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5161 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5221 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5281 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5341 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5401 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5461 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5521 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5581 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5641 tgactgcccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5701 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5761 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5821 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5881 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5941 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgaaaaaa agcggttagc
     6001 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6061 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6121 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6181 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6241 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6301 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6361 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6421 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6481 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6541 acatttcccc gaaaagtgcc acctgacgga tcgcttgcct gtaacttaca cgcgcctcgt
     6601 atcttttaat gatggaataa tttgggaatt tactctgtat ttatttattt ttatgttttg
     6661 tatttggatt ttagaaagta aataaagaag gtagaagagt tacggaatga agaaaaaaaa
     6721 ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt tacatatata tttattagac
     6781 aagaaaagca gattaaatag atatacattc gattaacgat aagtaaaatg taaaatcaca
     6841 ggattttcgt gtgtggtctt ctacacagac aagatgaaac aattcggcat taatacctga
     6901 gagcaggaag agcaagataa aaggtagtat ttgttggcga tccccctaga gtctt
//
