LOCUS       Exported                5772 bp DNA     circular SYN 14-JAN-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5772)
  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..5772
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     complement(17..39)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     139..158
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     171..226
                     /label=Primer 1
     promoter        177..843
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase"
     primer_bind     177..202
                     /label=Gib_SacI_GAProm_f
     primer_bind     177..202
                     /label=p16_GAPprom_f
     primer_bind     177..198
                     /label=NotI_GAPprom_f
     primer_bind     687..706
                     /label=GPD fw 1
     primer_bind     734..753
                     /label=GPD fw 2
     primer_bind     761..784
                     /label=p426-sense(GPD_pr)
     primer_bind     complement(804..849)
                     /label=Primer 2
     primer_bind     complement(819..844)
                     /label=mCherry_GAPprom_r
     CDS             879..1581
                     /codon_start=1
                     /label=yomTagRFP657
                     /translation="MSELITENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTQRIKV
                     VEGGPLPFAFDILATSFMYGSHTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEAHTEMLYPADGGLEGRTALALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDYRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     terminator      1601..1788
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1656..1680)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     1848..1878
                     /label=Primer 1
     misc_feature    1855..2358
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     1855..1875
                     /label=CEN/ARS_fwd
     primer_bind     complement(1855..1875)
                     /label=Gib_ARS_p16_rev
     primer_bind     complement(2354..2373)
                     /label=CEN/ARS_rev
     primer_bind     2354..2373
                     /label=Gib_ARS_p16_fwd
     primer_bind     2406..2424
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2462..2484)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2584..2603
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2748..2935
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             2936..3598
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     primer_bind     complement(3572..3604)
                     /label=Primer 2
     primer_bind     complement(3664..3683)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3665..3683)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3770..3787)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3941..4529)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4021..4040)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4700..5560)
                     /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     5323..5342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5561..5665)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5733..5751
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
       61 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
      121 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgtcat
      181 tatcaatact gccatttcaa agaatacgta aataattaat agtagtgatt ttcctaactt
      241 tatttagtca aaaaattagc cttttaattc tgctgtaacc cgtacatgcc caaaataggg
      301 ggcgggttac acagaatata taacatcgta ggtgtctggg tgaacagttt attcctggca
      361 tccactaaat ataatggagc ccgcttttta agctggcatc cagaaaaaaa aagaatccca
      421 gcaccaaaat attgttttct tcaccaacca tcagttcata ggtccattct cttagcgcaa
      481 ctacagagaa caggggcaca aacaggcaaa aaacgggcac aacctcaatg gagtgatgca
      541 acctgcctgg agtaaatgat gacacaaggc aattgaccca cgcatgtatc tatctcattt
      601 tcttacacct tctattacct tctgctctct ctgatttgga aaaagctgaa aaaaaaggtt
      661 gaaaccagtt ccctgaaatt attcccctac ttgactaata agtatataaa gacggtaggt
      721 attgattgta attctgtaaa tctatttctt aaacttctta aattctactt ttatagttag
      781 tctttttttt agttttaaaa caccaagaac ttagtttcga ataaacacac ataaacaaac
      841 aaagtcgacg gatcggtgac ggtgctggtt taattaacat gagcgaattg atcacagaaa
      901 atatgcacat gaagttgtat atggagggaa ccgtcaataa ccatcatttc aaatgcactt
      961 ctgaaggtga gggcaaacct tacgaaggga cccaaacaca gagaattaag gtagtagaag
     1021 ggggtccact gccatttgcc tttgacatcc tagcaacatc cttcatgtac ggttcccaca
     1081 cattcataaa ccacactcaa ggaataccag acttttggaa gcagagtttt ccagaaggtt
     1141 ttacgtggga aagagttact acttacgagg acggtggtgt attaacagcc acacaagata
     1201 catcattaca agatggttgt ctgatctaca acgttaaaat cagaggtgtt aactttcctt
     1261 caaatgggcc agttatgcaa aagaaaactt taggctggga ggcccatact gaaatgcttt
     1321 acccagcaga tggcggcttg gaaggcagga cggctttggc tttaaagcta gttggtggag
     1381 gacatttgat ttgtaatttc aaaacaactt acagaagcaa aaagcctgct aaaaatctta
     1441 agatgccagg agtgtactat gtcgattaca gattggaacg tattaaagag gctgataagg
     1501 aaacctatgt ggaacaacat gaagtggcag tcgcgagata ttgtgatctt ccttctaaac
     1561 taggtcataa actcaattaa ggcgcgccac ttctaaataa gcgaatttct tatgatttat
     1621 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     1681 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     1741 ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca gatccgctag
     1801 ggataacagg gtaatataga tctgtttagc ttgcctcgtc cccgccgggt caccgatcgc
     1861 ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg gaataatttg ggaatttact
     1921 ctgtatttat ttatttttat gttttgtatt tggattttag aaagtaaata aagaaggtag
     1981 aagagttacg gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc
     2041 gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat acattcgatt
     2101 aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac acagacaaga
     2161 tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg tagtatttgt
     2221 tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt ttctttaatt
     2281 tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta aattataatt
     2341 atttttatag cacgtgatga aaaggaccct aagaaaccat tattatcatg acattaacct
     2401 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     2461 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     2521 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg acgtcggggc tggcttaact
     2581 atgcggcatc agagcagatt gtactgagag tgcaccataa ttccgtttta agagcttggt
     2641 gagcgctagg agtcactgcc aggtatcgtt tgaacacggc attagtcagg gaagtcataa
     2701 cacagtcctt tcccgcaatt ttctttttct attactcttg gcctcctcta gtacactcta
     2761 tattttttta tgcctcggta atgattttca tttttttttt tccacctagc ggatgactct
     2821 ttttttttct tagcgattgg cattatcaca taatgaatta tacattatat aaagtaatgt
     2881 gatttcttcg aagaatatac taaaaaatga gcaggcaaga taaacgaagg caaagatgac
     2941 agagcagaaa gccctagtaa agcgtattac aaatgaaacc aagattcaga ttgcgatctc
     3001 tttaaagggt ggtcccctag cgatagagca ctcgatcttc ccagaaaaag aggcagaagc
     3061 agtagcagaa caggccacac aatcgcaagt gattaacgtc cacacaggta tagggtttct
     3121 ggaccatatg atacatgctc tggccaagca ttccggctgg tcgctaatcg ttgagtgcat
     3181 tggtgactta cacatagacg accatcacac cactgaagac tgcgggattg ctctcggtca
     3241 agcttttaaa gaggccctag gggccgtgcg tggagtaaaa aggtttggat caggatttgc
     3301 gcctttggat gaggcacttt ccagagcggt ggtagatctt tcgaacaggc cgtacgcagt
     3361 tgtcgaactt ggtttgcaaa gggagaaagt aggagatctc tcttgcgaga tgatcccgca
     3421 ttttcttgaa agctttgcag aggctagcag aattaccctc cacgttgatt gtctgcgagg
     3481 caagaatgat catcaccgta gtgagagtgc gttcaaggct cttgcggttg ccataagaga
     3541 agccacctcg cccaatggta ccaacgatgt tccctccacc aaaggtgttc ttatgtagga
     3601 gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg gatctgccgg
     3661 tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg
     3721 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3781 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3841 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3901 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3961 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4021 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4081 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     4141 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4201 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4261 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4321 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4381 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4441 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4501 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4561 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4621 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4681 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4741 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4801 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4861 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4921 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4981 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5041 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5101 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5161 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5221 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5281 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5341 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5401 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5461 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5521 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5581 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5641 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5701 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5761 tcgcgcgttt cg
//
