LOCUS       Exported                7543 bp ds-DNA     circular SYN 06-JUN-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    PmCHS_p426TEF
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7543)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 19, 2017 from SnapGene 3.3.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7543
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     rep_origin      69..1411
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    401..448
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        1438..1542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1543..2403
                     /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"
     rep_origin      2574..3162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3486..3516
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3524..3540
                     /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)."
     primer_bind     3548..3564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3585..3603
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3621..4019
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     4023..4029
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4035..5219
                     /codon_start=1
                     /product="Formerly called PmCHS3"
                     /label=PmCHS
                     /translation="MSKTVEEIWAAQRARGPATVLAIGTAAPANVVYQADYPDYYFRIT
                     KSEHMTELKEKFRRMCDKSMITKRHMHLSEELLKNNPDICAYMAPSLDARQDMVVVEVP
                     KLGKEAAAKAIKEWGRPKSAITHLIFCTTSGVDMPGADFQLTKLLGLCPSVRRTMLYQQ
                     GCFAGGTVLRLAKDLAENNAGARVLVVCSEITAVTFRGPSETHLDSMVGQALFGDGASA
                     IIVGADPDPVIERPLFQIVSAAQTILPDSDGAIDGHLREVGLTFHLLKDVPGLISKNIE
                     KSLKEAFAPLGIDDWNSIFWIVHPGGPAILDQVEAKLRLKVEKLKTTRTVLSEYGNMSS
                     ACVLFILDEMRRNSMEEGKATTGEGLHWGVLFGFGPGLTVETVVLHSLPIAEAN"
     primer_bind     5225..5230
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5264..5280)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5283..5530
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(5549..5567)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5577..5593)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5738..6193
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(6324..7127)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(7128..7343)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagtatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg
      121 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg
      181 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac
      241 gcatataagt acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata
      301 tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc
      361 gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt
      421 cctattctct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa
      481 gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga
      541 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat
      601 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
      661 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta
      721 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag
      781 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc
      841 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct
      901 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt
      961 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg
     1021 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa
     1081 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa
     1141 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta
     1201 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt
     1261 acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt
     1321 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
     1381 gttctacaaa atgaagcaca gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc
     1441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2221 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2701 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3361 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     3421 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt acctcactca ttaggcaccc
     3481 caggctttac actttatgct tccggctcct atgttgtgtg gaattgtgag cggataacaa
     3541 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     3601 agggaacaaa agctggagct catagcttca aaatgtttct actccttttt tactcttcca
     3661 gattttctcg gactccgcgc atcgccgtac cacttcaaaa cacccaagca cagcatacta
     3721 aatttcccct ctttcttcct ctagggtgtc gttaattacc cgtactaaag gtttggaaaa
     3781 gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa aatttttatc
     3841 acgtttcttt ttcttgaaaa tttttttttt gatttttttc tctttcgatg acctcccatt
     3901 gatatttaag ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc
     3961 tattacaact ttttttactt cttgctcatt agaaagaaag catagcaatc taatctaagt
     4021 tttctagaac tagtatgtcg aagaccgtag aggagatttg ggcggcgcag cgggcgaggg
     4081 gaccagccac ggtgctggcc atcggcactg ctgcgccggc caatgtggtg taccaggccg
     4141 attatccgga ctactacttt aggatcacca agagcgagca catgacagag ctcaaggaga
     4201 agttccgacg aatgtgtgac aagtcgatga taacgaagcg gcacatgcac ttgtcggagg
     4261 agctgttgaa aaacaaccct gacatctgtg cctacatggc cccttccctc gacgcccgcc
     4321 aagatatggt cgtggtggag gtacccaagc tcggcaagga ggcggccgcc aaggccatca
     4381 aggaatgggg ccgcccaaag tcggccatca cccacctcat cttctgcacc acctccggcg
     4441 tcgacatgcc cggcgccgat ttccagctca ccaagctact cggcctctgc ccctccgttc
     4501 gccgcaccat gctctaccag cagggctgct tcgccggcgg tacggttctc cgccttgcca
     4561 aggacctcgc cgagaacaat gcgggcgcga gggtcctcgt cgtctgctcc gagatcaccg
     4621 ccgtcacctt ccgcggcccc tcggagactc acctcgatag catggtcggc caggccctgt
     4681 tcggtgatgg tgcctctgcc atcatcgtcg gtgccgaccc cgaccccgtc atagaaaggc
     4741 cactctttca aattgtatct gcggctcaga ccatccttcc cgactcggat ggggcaatag
     4801 acggccatct ccgagaagtg ggtctaacct tccacctcct caaggacgta cctgggctca
     4861 tctcaaagaa catcgagaag agcctaaagg aggcgtttgc accgctgggc atcgacgact
     4921 ggaactcgat attttggatt gttcatccag gcgggccggc cattctagac caggtggagg
     4981 cgaagctgcg tctgaaagtg gagaagctga agacaacgag aacagttttg agtgagtacg
     5041 ggaatatgtc gagcgcttgc gtgttgttca tacttgacga gatgaggagg aacagcatgg
     5101 aagaagggaa ggcgacgacc ggtgaagggt tacattgggg agttttgttt ggttttgggc
     5161 cgggcttgac agtggagacg gtcgtcttgc atagtttgcc catcgccgag gccaactaaa
     5221 ctagtggatc ccccgggctg caggaattcg atatcaagct tatcgatacc gtcgacctcg
     5281 agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac
     5341 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
     5401 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
     5461 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc
     5521 tttaatttgc ggccggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     5581 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     5641 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     5701 tcccaacagt tgcgcagcct gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta
     5761 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5821 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5881 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5941 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     6001 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     6061 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     6121 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     6181 acgtttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     6241 gcatagggta ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat
     6301 gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc
     6361 cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt
     6421 cctcttccaa caataataat gtcagatcct gtagagacca catcatccac ggttctatac
     6481 tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa
     6541 tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct
     6601 ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc
     6661 ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc
     6721 gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct
     6781 gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg
     6841 tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc
     6901 ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa
     6961 caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc
     7021 aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca
     7081 ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt
     7141 ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct
     7201 tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc
     7261 ttctgttcgg agattaccga atcaaaaaaa tttcaaagaa accgaaatca aaaaaaagaa
     7321 taaaaaaaaa atgatgaatt gaattgaaaa gctgtggtat ggtgcactct cagtacaatc
     7381 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     7441 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     7501 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cga
//
