LOCUS       Exported                5397 bp ds-DNA     circular SYN 07-JAN-2025
DEFINITION  Plasmid for tagging Sec26 with HaloTag by gene replacement.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5397)
  AUTHORS   Adam Krahn
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 2, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Linearize with AflII for integration at SEC26.
FEATURES             Location/Qualifiers
     source          1..5397
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     CDS             55..363
                     /codon_start=1
                     /label=SEC26
                     /translation="VKSQLPTLHAYLRELVKGTNMGILTPSESLGEDDCRFLSCNLYAK
                     SSFGEDALANLCIEKDSKTNDVIGYVRIRSKGQGLALSLGDRVALIAKKTNKLALTH"
     CDS             371..401
                     /codon_start=1
                     /label=GSS linker
                     /translation="GSSGSSGSSG"
     CDS             407..1297
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /note="yeast codon-optimized"
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     misc_feature    1316..1976
                     /label=3' SEC26
     CDS             complement(2374..3177)
                     /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(3178..3398)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        3491..3595
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3596..4456
                     /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      4627..5215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagggtcaa
       61 atcacaacta ccaacattgc atgcttattt gagagaactg gtcaagggaa ctaatatggg
      121 tattctaaca ccatcagagt cgttgggaga agatgattgt aggttcttaa gttgtaatct
      181 gtatgcgaag tcgtcctttg gtgaagatgc cctagccaac ttgtgtatcg aaaaggattc
      241 caaaaccaat gatgtcatag gttatgttcg tatccgatca aagggacaag gtttggctct
      301 gtccctaggt gacagagtgg cattgattgc taagaagacc aataaacttg ctctcactca
      361 tgtggatcca ggaagttcgg ggtccagcgg ttcttcaggc gctagcatgg cagaaatcgg
      421 tacaggtttc ccttttgacc cacactatgt cgaagtatta ggtgaaagaa tgcactatgt
      481 tgatgtcggt cctagagatg gtactccagt tttgttttta catggtaacc caacatcttc
      541 atacgtttgg agaaacatca tcccacatgt tgcaccaact catagatgta ttgctccaga
      601 tttgattggt atgggtaaat ctgataagcc agatttgggt tatttctttg atgatcatgt
      661 tagattcatg gatgctttta ttgaagcatt gggtttggaa gaagttgttt tggttattca
      721 tgattggggt tcagcattag gttttcattg ggctaagaga aacccagaaa gagttaaggg
      781 tatcgctttt atggaattca ttagaccaat tccaacttgg gatgaatggc cagaattcgc
      841 aagagaaaca ttccaagctt ttagaactac agatgttggt agaaagttga tcatcgatca
      901 aaacgttttt attgaaggta ctttgccaat gggtgttgtt agaccattaa cagaagttga
      961 aatggatcat tacagagaac catttttaaa cccagttgat agagaaccat tgtggagatt
     1021 tccaaatgaa ttaccaattg ctggtgaacc agcaaacatc gttgctttgg ttgaagaata
     1081 catggattgg ttacatcaat ctccagttcc aaagttgtta ttttggggta caccaggtgt
     1141 tttgattcca ccagcagaag ctgcaagatt ggctaagtca ttgccaaact gtaaggctgt
     1201 tgatattggt ccaggtttaa atttgttaca agaagataat ccagacttga taggtagtga
     1261 aatagccaga tggttgtcca cattagaaat ctcaggttaa tctagaagcg gccgcaccac
     1321 cctgagctga tgcaatgtaa gttataatgt caaaggagat aagaataaaa cttaaaaaat
     1381 tcgaaaagaa ataaaaaaat aaggaaaatc tttaaataga ttttaattgc gtaggaatct
     1441 ttgttctcat ttttaaacaa gtattcactt ttatgtacat tttctttcct aaaaatcttt
     1501 atacagagtt gagttgtgag ttgtgtaaga gacgtccaat tttataacaa tttgtagaca
     1561 tctcagtgcc ccaaagagaa ctatatgatt gaaaaatgga gagagatagt atcttgtaca
     1621 gtttatatat ttacatgacg aatgggaaat tatatatgcc ctttgccttt ctgccataga
     1681 gcatggctaa ctggccagga ccatgagatt gtattcttat ttctcctctc cgaaaaatgg
     1741 gaactggaac ccactcaaca aagttctcgc ctggtaatct aattgagctg aggtgtttat
     1801 atttatgtgc ataccaaaag ttttaggcca tgagtcctgg tttgcatcga tttcagggaa
     1861 gaatttgata tcttcagccg tcaggccgaa actaatccca ccaaatctgt tcccagactg
     1921 gttgcttata cctttatatt ctctgattct tggtaggacg atttcggtca atgtgcctgg
     1981 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2041 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2101 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     2161 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     2221 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     2281 gcgagacgaa agggcgggta ataactgata taattaaatt gaagctctaa tttgtgagtt
     2341 tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa
     2401 atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt
     2461 tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca catcatccac
     2521 ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat
     2581 aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat
     2641 aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga
     2701 tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac
     2761 ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt
     2821 cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt
     2881 attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa
     2941 tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt
     3001 ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt
     3061 acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt
     3121 ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat
     3181 ttatcttcgt ttcctgcatg tttttgttct gtgcagttgg gttaagaata ctgggcaatt
     3241 tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct
     3301 tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaagaa accgaaatca
     3361 aaaaaaagaa taaaaaaaaa atgatgaatt gaattgaaaa gctggcctcg tgatacgcct
     3421 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3481 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3541 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3601 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3661 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3721 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3781 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3841 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3901 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3961 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4021 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4081 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4141 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4201 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4261 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4321 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4381 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4441 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4501 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4561 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4621 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4681 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4741 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4801 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4861 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4921 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4981 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5041 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5101 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5161 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5221 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5281 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5341 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaaga
//
