LOCUS       sequence-200156-        8262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  I149A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8262)
  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 8262)
  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 Ret2 I149A
FEATURES             Location/Qualifiers
     source          1..8262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(214..233)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     333..355
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(393..411)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    453..956
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        993..1097
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1098..1166,1167..1958)
                     /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(1316..1335)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2129..2717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2618..2637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2871..2888
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3005..3026
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3041..3046,3047..3064,3065..3071)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3079..3095
                     /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     3084..3106
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3103..3119
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3103..3119
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3138..3158
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3140..3158
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3209..3558
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3559..3575
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3559..3575
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5254..5270)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5255..5271)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5273..5520
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5289..5307)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5538..5557)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5539..5557)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5567..5584)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5567..5583)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5576..5598)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5724..6179
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5811..5830)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6021..6042
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6480..6887
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6888..7982
                     /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 atcatgaaac tatgattcct aatttatata ttggaggatt ttctctaaaa aaaaaaaaat
       61 acaacaaata aaaaacactc aatgacctga ccatttgatg gagtttaagt caataccttc
      121 ttgaaccatt tcccataatg gtgaaagttc cctcaagaat tttactctgt cagaaacggc
      181 cttaacgacg tagtcgatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
      241 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
      301 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
      361 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
      421 taatgtcatg ataataatgg tttcttagga cggatcgctt gcctgtaact tacacgcgcc
      481 tcgtatcttt taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt
      541 tttgtatttg gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa
      601 aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt
      661 agacaagaaa agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat
      721 cacaggattt tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac
      781 ctgagagcag gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt
      841 tacatcttcg gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt
      901 aatttatata tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa
      961 aggacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1021 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1081 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1141 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1201 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1261 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1321 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1381 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1441 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1501 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1561 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1621 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     1681 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     1741 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     1801 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     1861 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1921 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     1981 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2041 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2101 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2161 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2221 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2281 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2341 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2401 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2461 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2521 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2581 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2641 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2701 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2761 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2821 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2881 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2941 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     3001 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     3061 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     3121 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctccgga
     3181 tgcaagggtt cgaatccctt agctctcatt attttttgct ttttctcttg aggtcacatg
     3241 atcgcaaaat ggcaaatggc acgtgaagct gtcgatattg gggaactgtg gtggttggca
     3301 aatgactaat taagttagtc aaggcgccat cctcatgaaa actgtgtaac ataataaccg
     3361 aagtgtcgaa aaggtggcac cttgtccaat tgaacacgct cgatgaaaaa aataagatat
     3421 atataaggtt aagtaaagcg tctgttagaa aggaagtttt tcctttttct tgctctcttg
     3481 tcttttcatc tactatttcc ttcgtgtaat acagggtcgt cagatacata gatacaattc
     3541 tattaccccc atccatactc tagaactagt ggatccatgg tggttttggc tgcttctatc
     3601 actacgcgtc aggggaaacc tctcttgtcc aggcaattca aagatctgtc caaggatcga
     3661 gtcctagagt tgctgtccaa tttccagaac ctagtctccg agatctcgtc agaccacaca
     3721 tttgtcgagg acaagcacgt ccgctacgtg tataggccct ttgataacta ctacattatt
     3781 cttatcacaa accgccagtc caatattatc aaggatttgg ccacattaaa tctgttttca
     3841 cagacgatca actcctactt gtctagtttc caagaccaag agatttttca caatgccttt
     3901 gagatcttgt cctcgttcga cgaaatcgtg tcaatgggcg ggtacaaaga gaacttgtcc
     3961 tttactcaag tgcaaacata cttgtctatg gagtcgcacg aagaaagaat ccaggaaatt
     4021 gccgagcgta ataaagagat cgaagctact gaggagagga agcgccgtgc taaggaaatt
     4081 gctagaaaag aacatgaaag gaaacacggg tttatgtctt caaatggcga ttacgacggc
     4141 gccaataggt tcatgggaag taaagatcct aacgttacaa acgccatcaa cagttactac
     4201 tcccatgctt ctcctgcagc acaacagtct tatctacaaa gttcccatgc cgctgctgct
     4261 gaggtggccc cagtggcctc cccaatggct acgtctcagc gtgcgggaca ttctgctact
     4321 ggcggtatga aattgggtgg tggtgccgga agaagagcgg gtgccgcacc acggccaagt
     4381 gcaatttctt cagcctcttc tggcactcca cctcctccgg aggaggatgt cccagagaac
     4441 aatggtattt tgatctctat caaggaggta atcaatgcag aattctccag agatggtacc
     4501 attcactctt ctgaattgaa aggtgtttta gaactaagaa ttaatgacca tgacttatct
     4561 cactcgaacc taaaactagc tgattctatt gacgttcgcg ataaatcctt ccagtttaag
     4621 acccatccaa acatagataa acaatcgttc ctatccacta aacttatctc cctgcgtgat
     4681 aaatctaagg ctttcccagc caacgaccaa agcctgggtg ttctaagatg gcgtaaagtc
     4741 gctcccgcag aagatgactc attaattcca ttaactctaa ctacatgggt ttccccctcc
     4801 gaatcacaac agggctttga tgtcattatt gagtatgaga gcgtcctgga aacagaactc
     4861 gctgatgtga tcttcaccat tccagttttc cctcaggagc cagtagacat aaatacagag
     4921 tcttctacat gctctgacgc tgaagttgtc aacatggacc aagaaatggg aacctccatt
     4981 aagattagca aaattgctgc caacgatgca ggtgcgctag cattcactat cgaagctcca
     5041 tatgaagatg cattgtaccc aatgaccgtt tccttccaag agtccacgag ggataaatta
     5101 gctaagagtt tcaccggtat ggccatacaa tctgttgtca tggccaatga ccacgatcaa
     5161 gagctccctt acgacgtcat cacctctttg aaatcagatg aatatcttgt ccaatagtcc
     5221 ccccgggctg caggaattcg atatcaagct tatcgatacc gtcgacctcg agtcatgtaa
     5281 ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa
     5341 ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta
     5401 agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt
     5461 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc
     5521 ggccggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     5581 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     5641 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     5701 tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5761 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5821 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5881 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5941 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6001 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     6061 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6121 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc
     6181 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atcgaccggt
     6241 cgaggagaac ttctagtata tctacatacc taatattatt gccttattaa aaatggaatc
     6301 ccaacaatta catcaaaatc cacattctct tcaaaatcaa ttgtcctgta cttccttgtt
     6361 catgtgtgtt caaaaacgtt atatttatag gataattata ctctatttct caacaagtaa
     6421 ttggttgttt ggccgagcgg tctaaggcgc ctgattcaag aaatatcttg accgcagtta
     6481 actgtgggaa tactcaggta tcgtaagatg caagagttcg aatctcttag caaccattat
     6541 ttttttcctc aacataacga gaacacacag gggcgctatc gcacagaatc aaattcgatg
     6601 actggaaatt ttttgttaat ttcagaggtc gcctgacgca tatacctttt tcaactgaaa
     6661 aattgggaga aaaaggaaag gtgagagcgc cggaaccggc ttttcatata gaatagagaa
     6721 gcgttcatga ctaaatgctt gcatcacaat acttgaagtt gacaatatta tttaaggacc
     6781 tattgttttt tccaataggt ggttagcaat cgtcttactt tctaactttt cttacctttt
     6841 acatttcagc aatatatata tatatatttc aaggatatac cattctaatg tctgccccta
     6901 agaagatcgt cgttttgcca ggtgaccacg ttggtcaaga aatcacagcc gaagccatta
     6961 aggttcttaa agctatttct gatgttcgtt ccaatgtcaa gttcgatttc gaaaatcatt
     7021 taattggtgg tgctgctatc gatgctacag gtgttccact tccagatgag gcgctggaag
     7081 cctccaagaa ggctgatgcc gttttgttag gtgctgtggg tggtcctaaa tggggtaccg
     7141 gtagtgttag acctgaacaa ggtttactaa aaatccgtaa agaacttcaa ttgtacgcca
     7201 acttaagacc atgtaacttt gcatccgact ctcttttaga cttatctcca atcaagccac
     7261 aatttgctaa aggtactgac ttcgttgttg tcagagaatt agtgggaggt atttactttg
     7321 gtaagagaaa ggaagacgat ggtgatggtg tcgcttggga tagtgaacaa tacaccgttc
     7381 cagaagtgca aagaatcaca agaatggccg ctttcatggc cctacaacat gagccaccat
     7441 tgcctatttg gtccttggat aaagctaatg ttttggcctc ttcaagatta tggagaaaaa
     7501 ctgtggagga aaccatcaag aacgaattcc ctacattgaa ggttcaacat caattgattg
     7561 attctgccgc catgatccta gttaagaacc caacccacct aaatggtatt ataatcacca
     7621 gcaacatgtt tggtgatatc atctccgatg aagcctccgt tatcccaggt tccttgggtt
     7681 tgttgccatc tgcgtccttg gcctctttgc cagacaagaa caccgcattt ggtttgtacg
     7741 aaccatgcca cggttctgct ccagatttgc caaagaataa ggtcaaccct atcgccacta
     7801 tcttgtctgc tgcaatgatg ttgaaattgt cattgaactt gcctgaagaa ggtaaggcca
     7861 ttgaagatgc agttaaaaag gttttggatg caggtatcag aactggtgat ttaggtggtt
     7921 ccaacagtac caccgaagtc ggtgatgctg tcgccgaaga agttaagaaa atccttgctt
     7981 aaaaagattc tcttttttta tgatatttgt acataaactt tataaatgaa attcataata
     8041 gaaacgacac gaaattacaa aatggaatat gttcataggg tagacgaaac tatatacgca
     8101 atctacatac atttatcaag aaggagaaaa aggaggatgt aaaggaatac aggtaagcaa
     8161 attgatacta atggctcaac gtgataagga aaaagaattg cactttaaca ttaatattga
     8221 caaggaggag ggcaccacac aaaaagttag gtgtaacaga aa
//
