LOCUS       sequence-200182-        7131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  K153Q mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7131)
  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 7131)
  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 K153Q
FEATURES             Location/Qualifiers
     source          1..7131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(77..96)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     196..218
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(256..274)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    316..819
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        856..960
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(961..1029,1030..1821)
                     /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(1179..1198)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1992..2580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2481..2500
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2734..2751
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2868..2889
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2904..2909,2910..2927,2928..2934)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2942..2958
                     /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     2947..2969
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2966..2982
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2966..2982
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3001..3021
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3003..3021
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3072..3421
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3422..3438
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3422..3438
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3974..3991
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4005..4252
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4021..4039)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4270..4289)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4271..4289)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4299..4316)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4299..4315)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4308..4330)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4456..4911
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4543..4562)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4753..4774
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5212..5619
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5620..6714
                     /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 atggtgaaag ttccctcaag aattttactc tgtcagaaac ggccttaacg acgtagtcga
       61 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
      121 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
      181 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
      241 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
      301 tggtttctta ggacggatcg cttgcctgta acttacacgc gcctcgtatc ttttaatgat
      361 ggaataattt gggaatttac tctgtgttta tttattttta tgttttgtat ttggatttta
      421 gaaagtaaat aaagaaggta gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt
      481 taaaaaattt caacaaaaag cgtactttac atatatattt attagacaag aaaagcagat
      541 taaatagata tacattcgat taacgataag taaaatgtaa aatcacagga ttttcgtgtg
      601 tggtcttcta cacagacaag atgaaacaat tcggcattaa tacctgagag caggaagagc
      661 aagataaaag gtagtatttg ttggcgatcc ccctagagtc ttttacatct tcggaaaaca
      721 aaaactattt tttctttaat ttcttttttt actttctatt tttaatttat atatttatat
      781 taaaaaattt aaattataat tatttttata gcacgtgatg aaaaggaccc aggtggcact
      841 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
      901 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
      961 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1021 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     1081 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     1141 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     1201 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     1261 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     1321 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1381 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1441 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     1501 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     1561 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     1621 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     1681 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     1741 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     1801 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     1861 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     1921 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1981 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2041 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2101 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     2161 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2221 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2281 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2341 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2401 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2461 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2521 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2581 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2641 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     2701 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     2761 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     2821 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     2881 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     2941 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
     3001 gcaattaacc ctcactaaag ggaacaaaag ctggagctcc ggatgcaagg gttcgaatcc
     3061 cttagctctc attatttttt gctttttctc ttgaggtcac atgatcgcaa aatggcaaat
     3121 ggcacgtgaa gctgtcgata ttggggaact gtggtggttg gcaaatgact aattaagtta
     3181 gtcaaggcgc catcctcatg aaaactgtgt aacataataa ccgaagtgtc gaaaaggtgg
     3241 caccttgtcc aattgaacac gctcgatgaa aaaaataaga tatatataag gttaagtaaa
     3301 gcgtctgtta gaaaggaagt ttttcctttt tcttgctctc ttgtcttttc atctactatt
     3361 tccttcgtgt aatacagggt cgtcagatac atagatacaa ttctattacc cccatccata
     3421 ctctagaact agtggatcca tggtggtttt ggctgcttct atcactacgc gtcaggggaa
     3481 acctctcttg tccaggcaat tcaaagatct gtccaaggat cgagtcctag agttgctgtc
     3541 caatttccag aacctagtct ccgagatctc gtcagaccac acatttgtcg aggacaagca
     3601 cgtccgctac gtgtataggc cctttgataa ctactacatt attcttatca caaaccgcca
     3661 gtccaatatt atcaaggatt tggccacatt aaatctgttt tcacagacga tcaactccta
     3721 cttgtctagt ttccaagacc aagagatttt tcacaatgcc tttgagatct tgtcctcgtt
     3781 cgacgaaatc gtgtcaatgg gcgggtacaa agagaacttg tcctttactc aagtgcaaac
     3841 atacttgtct atggagtcgc acgaagaaag aatccaggaa attattgagc gtaatcaaga
     3901 gatcgaagct actgaggaga ggaagcgccg tgctaaggaa attgctagaa aagaacatga
     3961 aaggaaacac gggcatcacc atcaccatca ctagcccgct cgagtcatgt aattagttat
     4021 gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag
     4081 acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt
     4141 atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat
     4201 actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt gcggccggta
     4261 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     4321 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     4381 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     4441 ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4501 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4561 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4621 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4681 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4741 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     4801 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4861 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     4921 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatcgaccg gtcgaggaga
     4981 acttctagta tatctacata cctaatatta ttgccttatt aaaaatggaa tcccaacaat
     5041 tacatcaaaa tccacattct cttcaaaatc aattgtcctg tacttccttg ttcatgtgtg
     5101 ttcaaaaacg ttatatttat aggataatta tactctattt ctcaacaagt aattggttgt
     5161 ttggccgagc ggtctaaggc gcctgattca agaaatatct tgaccgcagt taactgtggg
     5221 aatactcagg tatcgtaaga tgcaagagtt cgaatctctt agcaaccatt atttttttcc
     5281 tcaacataac gagaacacac aggggcgcta tcgcacagaa tcaaattcga tgactggaaa
     5341 ttttttgtta atttcagagg tcgcctgacg catatacctt tttcaactga aaaattggga
     5401 gaaaaaggaa aggtgagagc gccggaaccg gcttttcata tagaatagag aagcgttcat
     5461 gactaaatgc ttgcatcaca atacttgaag ttgacaatat tatttaagga cctattgttt
     5521 tttccaatag gtggttagca atcgtcttac tttctaactt ttcttacctt ttacatttca
     5581 gcaatatata tatatatatt tcaaggatat accattctaa tgtctgcccc taagaagatc
     5641 gtcgttttgc caggtgacca cgttggtcaa gaaatcacag ccgaagccat taaggttctt
     5701 aaagctattt ctgatgttcg ttccaatgtc aagttcgatt tcgaaaatca tttaattggt
     5761 ggtgctgcta tcgatgctac aggtgttcca cttccagatg aggcgctgga agcctccaag
     5821 aaggctgatg ccgttttgtt aggtgctgtg ggtggtccta aatggggtac cggtagtgtt
     5881 agacctgaac aaggtttact aaaaatccgt aaagaacttc aattgtacgc caacttaaga
     5941 ccatgtaact ttgcatccga ctctctttta gacttatctc caatcaagcc acaatttgct
     6001 aaaggtactg acttcgttgt tgtcagagaa ttagtgggag gtatttactt tggtaagaga
     6061 aaggaagacg atggtgatgg tgtcgcttgg gatagtgaac aatacaccgt tccagaagtg
     6121 caaagaatca caagaatggc cgctttcatg gccctacaac atgagccacc attgcctatt
     6181 tggtccttgg ataaagctaa tgttttggcc tcttcaagat tatggagaaa aactgtggag
     6241 gaaaccatca agaacgaatt ccctacattg aaggttcaac atcaattgat tgattctgcc
     6301 gccatgatcc tagttaagaa cccaacccac ctaaatggta ttataatcac cagcaacatg
     6361 tttggtgata tcatctccga tgaagcctcc gttatcccag gttccttggg tttgttgcca
     6421 tctgcgtcct tggcctcttt gccagacaag aacaccgcat ttggtttgta cgaaccatgc
     6481 cacggttctg ctccagattt gccaaagaat aaggtcaacc ctatcgccac tatcttgtct
     6541 gctgcaatga tgttgaaatt gtcattgaac ttgcctgaag aaggtaaggc cattgaagat
     6601 gcagttaaaa aggttttgga tgcaggtatc agaactggtg atttaggtgg ttccaacagt
     6661 accaccgaag tcggtgatgc tgtcgccgaa gaagttaaga aaatccttgc ttaaaaagat
     6721 tctctttttt tatgatattt gtacataaac tttataaatg aaattcataa tagaaacgac
     6781 acgaaattac aaaatggaat atgttcatag ggtagacgaa actatatacg caatctacat
     6841 acatttatca agaaggagaa aaaggaggat gtaaaggaat acaggtaagc aaattgatac
     6901 taatggctca acgtgataag gaaaaagaat tgcactttaa cattaatatt gacaaggagg
     6961 agggcaccac acaaaaagtt aggtgtaaca gaaaatcatg aaactatgat tcctaattta
     7021 tatattggag gattttctct aaaaaaaaaa aaatacaaca aataaaaaac actcaatgac
     7081 ctgaccattt gatggagttt aagtcaatac cttcttgaac catttcccat a
//
