LOCUS       sequence-200184-        7130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T158K mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7130)
  AUTHORS   Arakel EC, Richter KP, Clancy A, Schwappach B
  TITLE     delta-COP contains a helix C-terminal to its longin domain key to 
            COPI dynamics and function.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jun 21;113(25):6916-21. doi: 
            10.1073/pnas.1603544113. Epub 2016 Jun 13.
  PUBMED    27298352
REFERENCE   2  (bases 1 to 7130)
  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"; doi: 
            "10.1073/pnas.1603544113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-06-21- 21"; volume: "113"; issue: "25"; pages: "6916-21"
COMMENT     Sequence Label: p415 Ret2LD2a T158K
FEATURES             Location/Qualifiers
     source          1..7130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..25)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     125..147
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(185..203)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    245..748
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        785..889
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(890..958,959..1750)
                     /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(1108..1127)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1921..2509
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2410..2429
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2663..2680
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2797..2818
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2833..2838,2839..2856,2857..2863)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2871..2887
                     /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     2876..2898
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2895..2911
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2895..2911
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2930..2950
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2932..2950
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3001..3350
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3351..3367
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3351..3367
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3903..3920
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3933..4180
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3949..3967)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4198..4217)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4199..4217)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4227..4244)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4227..4243)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4236..4258)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4384..4839
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4471..4490)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4681..4702
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5140..5547
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5548..6642
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
ORIGIN
        1 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
       61 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
      121 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
      181 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
      241 gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg gaataatttg
      301 ggaatttact ctgtgtttat ttatttttat gttttgtatt tggattttag aaagtaaata
      361 aagaaggtag aagagttacg gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc
      421 aacaaaaagc gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat
      481 acattcgatt aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac
      541 acagacaaga tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg
      601 tagtatttgt tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt
      661 ttctttaatt tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta
      721 aattataatt atttttatag cacgtgatga aaaggaccca ggtggcactt ttcggggaaa
      781 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      841 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      901 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      961 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1021 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1081 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     1141 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1201 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1261 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1321 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1381 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1441 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1501 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1561 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     1621 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1681 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1741 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     1801 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1861 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1921 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1981 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2041 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2101 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2161 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2221 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2281 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2341 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2401 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2461 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2521 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2581 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2641 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     2701 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     2761 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     2821 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     2881 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     2941 tcactaaagg gaacaaaagc tggagctccg gatgcaaggg ttcgaatccc ttagctctca
     3001 ttattttttg ctttttctct tgaggtcaca tgatcgcaaa atggcaaatg gcacgtgaag
     3061 ctgtcgatat tggggaactg tggtggttgg caaatgacta attaagttag tcaaggcgcc
     3121 atcctcatga aaactgtgta acataataac cgaagtgtcg aaaaggtggc accttgtcca
     3181 attgaacacg ctcgatgaaa aaaataagat atatataagg ttaagtaaag cgtctgttag
     3241 aaaggaagtt tttccttttt cttgctctct tgtcttttca tctactattt ccttcgtgta
     3301 atacagggtc gtcagataca tagatacaat tctattaccc ccatccatac tctagaacta
     3361 gtggatccat ggtggttttg gctgcttcta tcactacgcg tcaggggaaa cctctcttgt
     3421 ccaggcaatt caaagatctg tccaaggatc gagtcctaga gttgctgtcc aatttccaga
     3481 acctagtctc cgagatctcg tcagaccaca catttgtcga ggacaagcac gtccgctacg
     3541 tgtataggcc ctttgataac tactacatta ttcttatcac aaaccgccag tccaatatta
     3601 tcaaggattt ggccacatta aatctgtttt cacagacgat caactcctac ttgtctagtt
     3661 tccaagacca agagattttt cacaatgcct ttgagatctt gtcctcgttc gacgaaatcg
     3721 tgtcaatggg cgggtacaaa gagaacttgt cctttactca agtgcaaaca tacttgtcta
     3781 tggagtcgca cgaagaaaga atccaggaaa ttattgagcg taataaagag atcgaagcta
     3841 aggaggagag gaagcgccgt gctaaggaaa ttgctagaaa agaacatgaa aggaaacacg
     3901 ggcatcacca tcaccatcac tagccgctcg agtcatgtaa ttagttatgt cacgcttaca
     3961 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     4021 ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat ttatatttca
     4081 aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt
     4141 gcttgagaag gttttgggac gctcgaaggc tttaatttgc ggccggtacc caattcgccc
     4201 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     4261 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     4321 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4381 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4441 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4501 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4561 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4621 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4681 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4741 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4801 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     4861 tacgcatctg tgcggtattt cacaccgcat atcgaccggt cgaggagaac ttctagtata
     4921 tctacatacc taatattatt gccttattaa aaatggaatc ccaacaatta catcaaaatc
     4981 cacattctct tcaaaatcaa ttgtcctgta cttccttgtt catgtgtgtt caaaaacgtt
     5041 atatttatag gataattata ctctatttct caacaagtaa ttggttgttt ggccgagcgg
     5101 tctaaggcgc ctgattcaag aaatatcttg accgcagtta actgtgggaa tactcaggta
     5161 tcgtaagatg caagagttcg aatctcttag caaccattat ttttttcctc aacataacga
     5221 gaacacacag gggcgctatc gcacagaatc aaattcgatg actggaaatt ttttgttaat
     5281 ttcagaggtc gcctgacgca tatacctttt tcaactgaaa aattgggaga aaaaggaaag
     5341 gtgagagcgc cggaaccggc ttttcatata gaatagagaa gcgttcatga ctaaatgctt
     5401 gcatcacaat acttgaagtt gacaatatta tttaaggacc tattgttttt tccaataggt
     5461 ggttagcaat cgtcttactt tctaactttt cttacctttt acatttcagc aatatatata
     5521 tatatatttc aaggatatac cattctaatg tctgccccta agaagatcgt cgttttgcca
     5581 ggtgaccacg ttggtcaaga aatcacagcc gaagccatta aggttcttaa agctatttct
     5641 gatgttcgtt ccaatgtcaa gttcgatttc gaaaatcatt taattggtgg tgctgctatc
     5701 gatgctacag gtgttccact tccagatgag gcgctggaag cctccaagaa ggctgatgcc
     5761 gttttgttag gtgctgtggg tggtcctaaa tggggtaccg gtagtgttag acctgaacaa
     5821 ggtttactaa aaatccgtaa agaacttcaa ttgtacgcca acttaagacc atgtaacttt
     5881 gcatccgact ctcttttaga cttatctcca atcaagccac aatttgctaa aggtactgac
     5941 ttcgttgttg tcagagaatt agtgggaggt atttactttg gtaagagaaa ggaagacgat
     6001 ggtgatggtg tcgcttggga tagtgaacaa tacaccgttc cagaagtgca aagaatcaca
     6061 agaatggccg ctttcatggc cctacaacat gagccaccat tgcctatttg gtccttggat
     6121 aaagctaatg ttttggcctc ttcaagatta tggagaaaaa ctgtggagga aaccatcaag
     6181 aacgaattcc ctacattgaa ggttcaacat caattgattg attctgccgc catgatccta
     6241 gttaagaacc caacccacct aaatggtatt ataatcacca gcaacatgtt tggtgatatc
     6301 atctccgatg aagcctccgt tatcccaggt tccttgggtt tgttgccatc tgcgtccttg
     6361 gcctctttgc cagacaagaa caccgcattt ggtttgtacg aaccatgcca cggttctgct
     6421 ccagatttgc caaagaataa ggtcaaccct atcgccacta tcttgtctgc tgcaatgatg
     6481 ttgaaattgt cattgaactt gcctgaagaa ggtaaggcca ttgaagatgc agttaaaaag
     6541 gttttggatg caggtatcag aactggtgat ttaggtggtt ccaacagtac caccgaagtc
     6601 ggtgatgctg tcgccgaaga agttaagaaa atccttgctt aaaaagattc tcttttttta
     6661 tgatatttgt acataaactt tataaatgaa attcataata gaaacgacac gaaattacaa
     6721 aatggaatat gttcataggg tagacgaaac tatatacgca atctacatac atttatcaag
     6781 aaggagaaaa aggaggatgt aaaggaatac aggtaagcaa attgatacta atggctcaac
     6841 gtgataagga aaaagaattg cactttaaca ttaatattga caaggaggag ggcaccacac
     6901 aaaaagttag gtgtaacaga aaatcatgaa actatgattc ctaatttata tattggagga
     6961 ttttctctaa aaaaaaaaaa atacaacaaa taaaaaacac tcaatgacct gaccatttga
     7021 tggagtttaa gtcaatacct tcttgaacca tttcccataa tggtgaaagt tccctcaaga
     7081 attttactct gtcagaaacg gccttaacga cgtagtcgat atggtgcact
//
