LOCUS       Exported                5489 bp DNA     circular SYN 09-AUG-2021
DEFINITION  Plasmid for chromosomal tagging of Ent5 with yeast codon-optimized 
            HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5489)
  AUTHORS   Glick Lab
  TITLE     Direct Submission
  JOURNAL   Exported Sep 20, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
COMMENT     Linearize with Bsu36I for pop-in integration at ENT5, then select 
            for pop-outs on 5-FOA.
FEATURES             Location/Qualifiers
     source          1..5489
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     CDS             245..548
                     /codon_start=2
                     /label=ENT5 (3' portion)
                     /translation="GNNDNKNNNINSNSDDDDDEFGDFQSETSPDTTAPKTSNSKIDDL
                     LDWDGPKSDTDTTAAAQTSLPFAEKKQQKARPQATKDKSKGNDAFSDLFSYSKSLV"
     CDS             549..566
                     /codon_start=1
                     /label=Linker
                     /translation="GSSGSS"
     CDS             567..1460
                     /codon_start=1
                     /product="yeast codon-optimized HaloTag"
                     /label=yo-HaloTag
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     misc_feature    1461..1963
                     /label=3' ENT5
     CDS             complement(2466..3269)
                     /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(3270..3490)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        3583..3687
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3688..4548
                     /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      4719..5307
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgctggtaa taatgacaac aaaaacaata atatcaacag caatagtgat gatgatgatg
      301 atgagtttgg cgacttccaa agcgaaacga gtcccgacac aacagcacct aaaacttcca
      361 atagtaaaat agatgatttg cttgactggg atggcccaaa gtcagacaca gataccaccg
      421 ctgctgcgca aacatcattg ccctttgcgg agaaaaaaca gcaaaaagcc cgtcctcagg
      481 ctacaaagga caagtcgaag ggaaatgatg cattttccga cttattctct tactccaagt
      541 cgttggtcgg cagcagtggt agtagcatgg cagaaatcgg tacaggtttc ccttttgacc
      601 cacactatgt cgaagtatta ggtgaaagaa tgcactatgt tgatgtcggt cctagagatg
      661 gtactccagt tttgttttta catggtaacc caacatcttc atacgtttgg agaaacatca
      721 tcccacatgt tgcaccaact catagatgta ttgctccaga tttgattggt atgggtaaat
      781 ctgataagcc agatttgggt tatttctttg atgatcatgt tagattcatg gatgctttta
      841 ttgaagcatt gggtttggaa gaagttgttt tggttattca tgattggggt tcagcattag
      901 gttttcattg ggctaagaga aacccagaaa gagttaaggg tatcgctttt atggaattca
      961 ttagaccaat tccaacttgg gatgaatggc cagaattcgc aagagaaaca ttccaagctt
     1021 ttagaactac agatgttggt agaaagttga tcatcgatca aaacgttttt attgaaggta
     1081 ctttgccaat gggtgttgtt agaccattaa cagaagttga aatggatcat tacagagaac
     1141 catttttaaa cccagttgat agagaaccat tgtggagatt tccaaatgaa ttaccaattg
     1201 ctggtgaacc agcaaacatc gttgctttgg ttgaagaata catggattgg ttacatcaat
     1261 ctccagttcc aaagttgtta ttttggggta caccaggtgt tttgattcca ccagcagaag
     1321 ctgcaagatt ggctaagtca ttgccaaact gtaaggctgt tgatattggt ccaggtttaa
     1381 atttgttaca agaagataat ccagacttga taggtagtga aatagccaga tggttgtcca
     1441 cattagaaat ctcaggttaa tcaccaaata ccccttcatc tcccaataca ttcaatacag
     1501 tcaacaaatt ttcattctca gatatatgta tccatatata cactaataat atcttactaa
     1561 aaacttgtct atttccttct tcaaatgtca gttttctttt tattgaatat caaaaaaata
     1621 ttatatagat acatttgtat atgtcattat ccatgcagaa gaaaatactt aaatacttaa
     1681 gtacacaagt atttattgat ttgctatttc ggtcttggca agtcccttgg ctacgtcatc
     1741 tacactcgcg tcatcctctt ccatatactc gtcgcctgtg cccaccacgc cttgttcaaa
     1801 catttgctta ccggttagct tcccatgatg agccttcatc cttctcacct ggtctctttc
     1861 atccagtttc agttcttggc gaaatttaga cctccattcc aagtacgatt ctcttgtgac
     1921 tttagtacca tggaacttgg cctgttcctt cttttcacgt tctggtaccg agctcgaatt
     1981 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     2041 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     2101 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     2161 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     2221 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     2281 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     2341 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcggg taataactga
     2401 tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca
     2461 gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa
     2521 cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata
     2581 atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct
     2641 cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt
     2701 ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg
     2761 tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct
     2821 aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta
     2881 acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg
     2941 tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct
     3001 tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc
     3061 atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat
     3121 gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt
     3181 gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca
     3241 gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca tgtttttgtt
     3301 ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt
     3361 atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc
     3421 gaatcaaaaa aatttcaaag aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa
     3481 ttgaattgaa aagctggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3541 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3601 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3661 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3721 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3781 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3841 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3901 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3961 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4021 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4081 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4141 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4201 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4261 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4321 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4381 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4441 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4501 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4561 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4621 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4681 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4741 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4801 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4861 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4921 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4981 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5041 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5101 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5161 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5221 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5281 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5341 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5401 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5461 agcgagtcag tgagcgagga agcggaaga
//
