LOCUS       sequence-348250-        9576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HaloTag-Drs2 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9576)
  AUTHORS   Casler JC, Johnson N, Krahn AH, Pantazopoulou A, Day KJ, Glick BS
  TITLE     Clathrin adaptors mediate two sequential pathways of intra-Golgi 
            recycling.
  JOURNAL   J Cell Biol. 2022 Jan 3;221(1). pii: 212747. doi: 
            10.1083/jcb.202103199. Epub 2021 Nov 5.
  PUBMED    34739034
REFERENCE   2  (bases 1 to 9576)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol."; date: "2022-01-3"; volume: "221(1). pii: 212747. doi"; 
            pages: " 10.1083/jcb.202103199. Epub 2021 Nov 5"
COMMENT     Sequence Label: YIplac211-HaloTag-DRS2
FEATURES             Location/Qualifiers
     source          1..9576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             880..1770
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     primer_bind     complement(6074..6091)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6074..6090)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6083..6105)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6299..6318)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6418..6440
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6558..7361)
                     /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(7362..7577)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7584..7602)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7670..7774
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7775..7843,7844..8635)
                     /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     complement(7993..8012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8806..9394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9295..9314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9548..9565
                     /label=L4440
                     /note="L4440 vector, forward primer"
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 atgcctgcag gtcgactcta gtgccaaaac taaacgtcct aaagtcgaaa tcgaatacga
      301 ggaggaacac gaagttcaaa atgctgagca agaagtggca caataagctc acgcactata
      361 ctacgcaaca ttacatgatt tcagtttcta ttattgaatg aacaggattt ttccacgtgg
      421 attgggcagc gcactgtacc ataataacaa ccagttataa aatatttcaa ttcttttaca
      481 tatataaaaa tacataaaga ctattatttt tgtaagatgg cccattctgt aacacttaaa
      541 tttcttatca aaaaaaaaaa aaataatacc tttagtagct gttgggctaa taacattgca
      601 aattagaagc ttactattag aagcttacta ttacgggatt attgaggtaa aggttatttg
      661 aagcgtaaat cactgaaatc ttcctgatca cgtgattggc tttttttgcc agaaaaccca
      721 aatctgttta atgccactaa gttgaagaaa catataatca acaattgttg gcaattctaa
      781 ggggaatagt gcaacaagaa gccagtaacg ccaaatagag gttagcgcca agcaataatt
      841 tactggtagg ttctttcatt ccctgatttt accatgaata tggcagaaat cggtacaggt
      901 ttcccttttg acccacacta tgtcgaagta ttaggtgaaa gaatgcacta tgttgatgtc
      961 ggtcctagag atggtactcc agttttgttt ttacatggta acccaacatc ttcatacgtt
     1021 tggagaaaca tcatcccaca tgttgcacca actcatagat gtattgctcc agatttgatt
     1081 ggtatgggta aatctgataa gccagatttg ggttatttct ttgatgatca tgttagattc
     1141 atggatgctt ttattgaagc attgggtttg gaagaagttg ttttggttat tcatgattgg
     1201 ggttcagcat taggttttca ttgggctaag agaaacccag aaagagttaa gggtatcgct
     1261 tttatggaat tcattagacc aattccaact tgggatgaat ggccagaatt cgcaagagaa
     1321 acattccaag cttttagaac tacagatgtt ggtagaaagt tgatcatcga tcaaaacgtt
     1381 tttattgaag gtactttgcc aatgggtgtt gttagaccat taacagaagt tgaaatggat
     1441 cattacagag aaccattttt aaacccagtt gatagagaac cattgtggag atttccaaat
     1501 gaattaccaa ttgctggtga accagcaaac atcgttgctt tggttgaaga atacatggat
     1561 tggttacatc aatctccagt tccaaagttg ttattttggg gtacaccagg tgttttgatt
     1621 ccaccagcag aagctgcaag attggctaag tcattgccaa actgtaaggc tgttgatatt
     1681 ggtccaggtt taaatttgtt acaagaagat aatccagact tgataggtag tgaaatagcc
     1741 agatggttgt ccacattaga aatctcaggt ggtagctcag ggtctagtgg atccgacgac
     1801 agagaaaccc ccccaaagag gaaacctggg gaggacgaca cgcttttcga tattgacttc
     1861 ttagacgaca cgacttctca ttctggatcc cgttcaaagg tcaccaactc acatgcgaac
     1921 gcgaattata taccaccaag tcatgtactt cccgaagaga ctatcgactt agacgctgat
     1981 gatgataaca ttgaaaatga tgtccatgaa aatctattta tgagcaataa ccatgacgac
     2041 caaacgtctt ggaatgccaa tagattcgac tcagatgcat atcaaccaca atccctacga
     2101 gcagtcaagc ctcccggttt atttgctagg tttggtaatg gtctcaaaaa tgcctttacg
     2161 ttcaaacgga agaaagggcc cgaaagtttc gaaatgaatc attacaatgc ggtgactaat
     2221 aacgaattag acgataatta tctagactcg aggaataagt tcaacattaa aattcttttc
     2281 aaccggtaca ttttaagaaa gaacgttggc gacgcagagg gcaatggtga accaagagtc
     2341 atccacatca acgattctct cgctaattcc tccttcggtt atagtgacaa ccatatatct
     2401 accaccaagt ataattttgc gacctttctg cctaaatttt tgttccaaga attttccaaa
     2461 tacgctaatc tgtttttttt atgtacatct gccattcaac aggtgcctca cgtctcgcca
     2521 actaatagat acaccacaat tggtacttta ttagtggttt tgattgtttc tgccatgaag
     2581 gaatgtatcg aagatatcaa gagagctaat tctgataaag aattaaataa ttcgacagca
     2641 gaaatatttt cagaagcaca tgatgacttt gttgagaaac gatggattga tattcgtgta
     2701 ggtgacataa ttagagtaaa atcagaggaa cctattcctg ctgataccat aattttgtca
     2761 tcttcggaac cggagggtct ttgctacatt gaaactgcca acttggatgg tgaaacaaat
     2821 ttgaaaatca aacagtcgag agtagaaact gccaaattta tagacgttaa aactttgaag
     2881 aacatgaacg gaaaagttgt ttccgaacag ccgaactcta gcttgtatac ttatgaaggt
     2941 acaatgactt taaatgatcg tcaaataccg ttatctcctg accaaatgat tttaagaggt
     3001 gcaactttaa gaaatactgc atggatattt ggtttagtta tcttcactgg tcatgaaact
     3061 aagttattgc gtaatgctac tgcaacccca attaaaagaa ccgcggttga gaaaattatc
     3121 aacagacaga ttattgcatt gttcacagtt ttaatcgtgc taattttaat ttcttcaatt
     3181 ggtaatgtta ttatgtctac tgcagatgcc aaacatttat cgtaccttta cctggagggt
     3241 accaacaagg ctggcttatt cttcaaagac tttttaacat tttggattct attttcgaat
     3301 ctagttccta tttctctatt tgtcaccgtt gaattaatca aatattatca ggcttttatg
     3361 ataggttcag atctagattt gtactacgaa aaaactgata ccccaactgt ggttcgtaca
     3421 tcctctttgg ttgaagaact tggtcaaata gaatatatat tcagtgacaa gacaggaact
     3481 ttaacaagaa atattatgga atttaaatcc tgctctattg caggccattg ctatattgat
     3541 aaaatacctg aagataaaac ggccactgtt gaagatggga ttgaagttgg ttatagaaaa
     3601 tttgacgatt tgaagaaaaa gttaaatgat ccctctgatg aggattcacc tattatcaat
     3661 gacttcttaa cattactggc tacctgtcat actgtcattc cagaatttca aagcgatgga
     3721 tctattaaat atcaagcagc ctctccagat gaaggtgccc tcgttcaagg tggtgcagat
     3781 ttagggtata agtttatcat ccgtaaacca aactctgtaa ctgttttatt ggaggaaact
     3841 ggcgaggaaa aagagtatca actacttaac atttgtgaat ttaattccac caggaagaga
     3901 atgagcgcta tatttagatt tccggatggt tcgataaagt tattctgtaa aggtgctgat
     3961 actgtcattc tggaaagatt ggatgatgaa gctaatcaat atgtagaagc tacaatgaga
     4021 catttagaag attatgcatc tgagggtttg cggacgttgt gtttggcgat gagagatatc
     4081 tctgaaggag aatatgaaga atggaatagc atttataatg aggctgccac aacattagat
     4141 aacagagccg agaagctaga cgaagccgca aatctaatcg agaaaaatct aatattaatt
     4201 ggtgcaactg ctattgaaga taagttacag gacggtgttc cagaaactat ccacacatta
     4261 caagaagcgg gtattaaaat ttgggtttta actggtgaca gacaggaaac tgctattaac
     4321 attggtatga gttgccgttt attgagtgaa gacatgaatt tgttgatcat caacgaggaa
     4381 accagagatg atactgagag aaatttgtta gagaagatta acgctctaaa cgagcatcaa
     4441 ttgtcaacac atgatatgaa taccttagcg ctcgtcattg atgggaagtc attaggcttt
     4501 gccttggaac cagaattaga agattatttg ctaactgtgg cgaagctttg taaagcggtt
     4561 atatgttgcc gtgtatctcc actacagaaa gccttggttg ttaaaatggt aaaaagaaag
     4621 tcgtcttcac tactgctagc cattggcgat ggtgccaacg atgttagtat gatacaggca
     4681 gctcatgttg gtgtcggtat tagtggtatg gaaggtatgc aagcggctcg ttcagctgat
     4741 atagctgttg gccaatttaa atttttaaaa aaactattac ttgtccatgg ctcctggtct
     4801 tatcaaagaa tttctgccgc aattttgtac tctttttaca agaatacagc attatacatg
     4861 acgcagtttt ggtacgtttt tgccaatgcc ttttcaggtc aatccattat ggaatcatgg
     4921 acaatgagtt tctacaactt attcttcact gtttggcccc cttttgtcat tggtgtattt
     4981 gatcaatttg ttagtagtag attacttgag cggtacccac aattgtacaa attaggacag
     5041 aagggtcaat ttttctctgt ttacatcttc tggggatgga ttattaatgg ctttttccat
     5101 tctgcaatag tatttattgg caccatattg atttacagat atgggtttgc cttaaatatg
     5161 cacggcgaac tagctgatca ttggtcttgg ggtgtaactg tctacactac aagtgtcatt
     5221 attgttttgg gaaaggctgc tttggtaact aatcaatgga cgaaattcac cttaattgcc
     5281 attcctggtt ccctcttatt ttggttaata ttcttcccaa tttatgcttc tatatttcct
     5341 catgctaaca tctcaagaga gtattatggt gtggttaaac acacgtatgg atccggtgta
     5401 ttttggttaa ctttgatcgt tttaccaatt tttgcactgg taagagattt tctatggaag
     5461 tactataaaa gaatgtatga accagaaacg tatcatgtta ttcaagaaat gcagaaatac
     5521 aatatcagcg actctaggcc gcacgttcag caatttcaaa atgccatcag gaaggtgagg
     5581 caagtgcaaa gaatgaaaaa acaaagagga tttgcctttt cacaggctga agagggtggg
     5641 caagaaaaaa ttgtcagaat gtatgatact actcaaaaga gaggtaagta cggtgaatta
     5701 caagatgctt cggcgaaccc ctttaatgac aataatggac tgggaagtaa cgactttgaa
     5761 agtgcagaac cgttcattga aaatccattt gctgatggta atcaaaattc aaatagattc
     5821 agttcttcga gagatgatat ttcatttgat atatgaggca cagaaacaaa gagcagaaga
     5881 caaaaaatac aaaaataaaa ataagataaa aagaaaagtc attattacca ttactgaaat
     5941 atgtacattt accataaaat atatccgtta tatagaagtt ataatattaa gccatcactt
     6001 tgttatcggt ttttcattcc tcctggggtt gcttcgactg ggtacgtacc ttatgtcttc
     6061 taggctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     6121 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     6181 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     6241 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     6301 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     6361 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     6421 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcg
     6481 ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat acatgcattt
     6541 acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct
     6601 gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctctt
     6661 ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct atactgttga
     6721 cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa
     6781 ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa atctttgtcg
     6841 ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga gcccttgcat
     6901 gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc tgccgcctgc
     6961 ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat gtctgcccat
     7021 tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc aatgtcagca
     7081 aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt tagcggctta
     7141 actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag taaacaaatt
     7201 ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac atccaatgaa
     7261 gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc aacaggacta
     7321 ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct tcgtttcctg
     7381 catgtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt ttcttcaaca
     7441 ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt
     7501 tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa
     7561 aaaaatgatg aattgaaaag ctggcctcgt gatacgccta tttttatagg ttaatgtcat
     7621 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     7681 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     7741 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     7801 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     7861 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     7921 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     7981 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     8041 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     8101 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     8161 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     8221 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     8281 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     8341 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     8401 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     8461 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     8521 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     8581 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     8641 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     8701 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     8761 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     8821 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     8881 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     8941 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     9001 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     9061 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     9121 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     9181 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     9241 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     9301 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     9361 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     9421 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     9481 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     9541 cgagcgcagc gagtcagtga gcgaggaagc ggaaga
//
