LOCUS       sequence-200157-        8262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  I155A 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 I155A
FEATURES             Location/Qualifiers
     source          1..8262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(211..230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     330..352
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(390..408)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    450..953
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        990..1094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1095..1163,1164..1955)
                     /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(1313..1332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2126..2714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2615..2634
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2868..2885
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3002..3023
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3038..3043,3044..3061,3062..3068)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3076..3092
                     /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     3081..3103
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3100..3116
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3100..3116
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3135..3155
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3137..3155
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3206..3555
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3556..3572
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3556..3572
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5251..5267)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5252..5268)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      5270..5517
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5286..5304)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5535..5554)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5536..5554)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5564..5581)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5564..5580)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5573..5595)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5721..6176
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5808..5827)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6018..6039
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6477..6884
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6885..7979
                     /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 atgaaactat gattcctaat ttatatattg gaggattttc tctaaaaaaa aaaaaataca
       61 acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa taccttcttg
      121 aaccatttcc cataatggtg aaagttccct caagaatttt actctgtcag aaacggcctt
      181 aacgacgtag tcgatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
      241 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
      301 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
      361 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
      421 tgtcatgata ataatggttt cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg
      481 tatcttttaa tgatggaata atttgggaat ttactctgtg tttatttatt tttatgtttt
      541 gtatttggat tttagaaagt aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa
      601 aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact ttacatatat atttattaga
      661 caagaaaagc agattaaata gatatacatt cgattaacga taagtaaaat gtaaaatcac
      721 aggattttcg tgtgtggtct tctacacaga caagatgaaa caattcggca ttaatacctg
      781 agagcaggaa gagcaagata aaaggtagta tttgttggcg atccccctag agtcttttac
      841 atcttcggaa aacaaaaact attttttctt taatttcttt ttttactttc tatttttaat
      901 ttatatattt atattaaaaa atttaaatta taattatttt tatagcacgt gatgaaaagg
      961 acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1021 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1081 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1141 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1201 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1261 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1321 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1381 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1441 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1501 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1561 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1621 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     1681 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1741 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1801 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1861 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1921 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1981 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2041 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2101 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2161 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2221 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2281 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2341 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2401 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2461 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2521 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2581 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2641 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2701 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2761 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2821 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2881 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2941 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     3001 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     3061 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     3121 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctccggatgc
     3181 aagggttcga atcccttagc tctcattatt ttttgctttt tctcttgagg tcacatgatc
     3241 gcaaaatggc aaatggcacg tgaagctgtc gatattgggg aactgtggtg gttggcaaat
     3301 gactaattaa gttagtcaag gcgccatcct catgaaaact gtgtaacata ataaccgaag
     3361 tgtcgaaaag gtggcacctt gtccaattga acacgctcga tgaaaaaaat aagatatata
     3421 taaggttaag taaagcgtct gttagaaagg aagtttttcc tttttcttgc tctcttgtct
     3481 tttcatctac tatttccttc gtgtaataca gggtcgtcag atacatagat acaattctat
     3541 tacccccatc catactctag aactagtgga tccatggtgg ttttggctgc ttctatcact
     3601 acgcgtcagg ggaaacctct cttgtccagg caattcaaag atctgtccaa ggatcgagtc
     3661 ctagagttgc tgtccaattt ccagaaccta gtctccgaga tctcgtcaga ccacacattt
     3721 gtcgaggaca agcacgtccg ctacgtgtat aggccctttg ataactacta cattattctt
     3781 atcacaaacc gccagtccaa tattatcaag gatttggcca cattaaatct gttttcacag
     3841 acgatcaact cctacttgtc tagtttccaa gaccaagaga tttttcacaa tgcctttgag
     3901 atcttgtcct cgttcgacga aatcgtgtca atgggcgggt acaaagagaa cttgtccttt
     3961 actcaagtgc aaacatactt gtctatggag tcgcacgaag aaagaatcca ggaaattatt
     4021 gagcgtaata aagaggccga agctactgag gagaggaagc gccgtgctaa ggaaattgct
     4081 agaaaagaac atgaaaggaa acacgggttt atgtcttcaa atggcgatta cgacggcgcc
     4141 aataggttca tgggaagtaa agatcctaac gttacaaacg ccatcaacag ttactactcc
     4201 catgcttctc ctgcagcaca acagtcttat ctacaaagtt cccatgccgc tgctgctgag
     4261 gtggccccag tggcctcccc aatggctacg tctcagcgtg cgggacattc tgctactggc
     4321 ggtatgaaat tgggtggtgg tgccggaaga agagcgggtg ccgcaccacg gccaagtgca
     4381 atttcttcag cctcttctgg cactccacct cctccggagg aggatgtccc agagaacaat
     4441 ggtattttga tctctatcaa ggaggtaatc aatgcagaat tctccagaga tggtaccatt
     4501 cactcttctg aattgaaagg tgttttagaa ctaagaatta atgaccatga cttatctcac
     4561 tcgaacctaa aactagctga ttctattgac gttcgcgata aatccttcca gtttaagacc
     4621 catccaaaca tagataaaca atcgttccta tccactaaac ttatctccct gcgtgataaa
     4681 tctaaggctt tcccagccaa cgaccaaagc ctgggtgttc taagatggcg taaagtcgct
     4741 cccgcagaag atgactcatt aattccatta actctaacta catgggtttc cccctccgaa
     4801 tcacaacagg gctttgatgt cattattgag tatgagagcg tcctggaaac agaactcgct
     4861 gatgtgatct tcaccattcc agttttccct caggagccag tagacataaa tacagagtct
     4921 tctacatgct ctgacgctga agttgtcaac atggaccaag aaatgggaac ctccattaag
     4981 attagcaaaa ttgctgccaa cgatgcaggt gcgctagcat tcactatcga agctccatat
     5041 gaagatgcat tgtacccaat gaccgtttcc ttccaagagt ccacgaggga taaattagct
     5101 aagagtttca ccggtatggc catacaatct gttgtcatgg ccaatgacca cgatcaagag
     5161 ctcccttacg acgtcatcac ctctttgaaa tcagatgaat atcttgtcca atagtccccc
     5221 cgggctgcag gaattcgata tcaagcttat cgataccgtc gacctcgagt catgtaatta
     5281 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
     5341 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     5401 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     5461 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc
     5521 cggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     5581 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     5641 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     5701 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5761 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5821 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5881 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5941 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6001 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     6061 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6121 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     6181 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatc gaccggtcga
     6241 ggagaacttc tagtatatct acatacctaa tattattgcc ttattaaaaa tggaatccca
     6301 acaattacat caaaatccac attctcttca aaatcaattg tcctgtactt ccttgttcat
     6361 gtgtgttcaa aaacgttata tttataggat aattatactc tatttctcaa caagtaattg
     6421 gttgtttggc cgagcggtct aaggcgcctg attcaagaaa tatcttgacc gcagttaact
     6481 gtgggaatac tcaggtatcg taagatgcaa gagttcgaat ctcttagcaa ccattatttt
     6541 tttcctcaac ataacgagaa cacacagggg cgctatcgca cagaatcaaa ttcgatgact
     6601 ggaaattttt tgttaatttc agaggtcgcc tgacgcatat acctttttca actgaaaaat
     6661 tgggagaaaa aggaaaggtg agagcgccgg aaccggcttt tcatatagaa tagagaagcg
     6721 ttcatgacta aatgcttgca tcacaatact tgaagttgac aatattattt aaggacctat
     6781 tgttttttcc aataggtggt tagcaatcgt cttactttct aacttttctt accttttaca
     6841 tttcagcaat atatatatat atatttcaag gatataccat tctaatgtct gcccctaaga
     6901 agatcgtcgt tttgccaggt gaccacgttg gtcaagaaat cacagccgaa gccattaagg
     6961 ttcttaaagc tatttctgat gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa
     7021 ttggtggtgc tgctatcgat gctacaggtg ttccacttcc agatgaggcg ctggaagcct
     7081 ccaagaaggc tgatgccgtt ttgttaggtg ctgtgggtgg tcctaaatgg ggtaccggta
     7141 gtgttagacc tgaacaaggt ttactaaaaa tccgtaaaga acttcaattg tacgccaact
     7201 taagaccatg taactttgca tccgactctc ttttagactt atctccaatc aagccacaat
     7261 ttgctaaagg tactgacttc gttgttgtca gagaattagt gggaggtatt tactttggta
     7321 agagaaagga agacgatggt gatggtgtcg cttgggatag tgaacaatac accgttccag
     7381 aagtgcaaag aatcacaaga atggccgctt tcatggccct acaacatgag ccaccattgc
     7441 ctatttggtc cttggataaa gctaatgttt tggcctcttc aagattatgg agaaaaactg
     7501 tggaggaaac catcaagaac gaattcccta cattgaaggt tcaacatcaa ttgattgatt
     7561 ctgccgccat gatcctagtt aagaacccaa cccacctaaa tggtattata atcaccagca
     7621 acatgtttgg tgatatcatc tccgatgaag cctccgttat cccaggttcc ttgggtttgt
     7681 tgccatctgc gtccttggcc tctttgccag acaagaacac cgcatttggt ttgtacgaac
     7741 catgccacgg ttctgctcca gatttgccaa agaataaggt caaccctatc gccactatct
     7801 tgtctgctgc aatgatgttg aaattgtcat tgaacttgcc tgaagaaggt aaggccattg
     7861 aagatgcagt taaaaaggtt ttggatgcag gtatcagaac tggtgattta ggtggttcca
     7921 acagtaccac cgaagtcggt gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa
     7981 aagattctct ttttttatga tatttgtaca taaactttat aaatgaaatt cataatagaa
     8041 acgacacgaa attacaaaat ggaatatgtt catagggtag acgaaactat atacgcaatc
     8101 tacatacatt tatcaagaag gagaaaaagg aggatgtaaa ggaatacagg taagcaaatt
     8161 gatactaatg gctcaacgtg ataaggaaaa agaattgcac tttaacatta atattgacaa
     8221 ggaggagggc accacacaaa aagttaggtg taacagaaaa tc
//
