LOCUS       sequence-200180-        7130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Q146F 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 Q146F
FEATURES             Location/Qualifiers
     source          1..7130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(384..403)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     503..525
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(563..581)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    623..1126
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        1163..1267
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1268..1336,1337..2128)
                     /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(1486..1505)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2299..2887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2788..2807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3041..3058
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3175..3196
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3211..3216,3217..3234,3235..3241)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3249..3265
                     /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     3254..3276
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3273..3289
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3273..3289
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3308..3328
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3310..3328
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3379..3728
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3729..3745
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3729..3745
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4281..4298
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4311..4558
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4327..4345)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4576..4595)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4577..4595)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4605..4622)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4605..4621)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4614..4636)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4762..5217
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4849..4868)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5059..5080
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5518..5925
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5926..7020
                     /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 tatacgcaat ctacatacat ttatcaagaa ggagaaaaag gaggatgtaa aggaatacag
       61 gtaagcaaat tgatactaat ggctcaacgt gataaggaaa aagaattgca ctttaacatt
      121 aatattgaca aggaggaggg caccacacaa aaagttaggt gtaacagaaa atcatgaaac
      181 tatgattcct aatttatata ttggaggatt ttctctaaaa aaaaaaaaat acaacaaata
      241 aaaaacactc aatgacctga ccatttgatg gagtttaagt caataccttc ttgaaccatt
      301 tcccataatg gtgaaagttc cctcaagaat tttactctgt cagaaacggc cttaacgacg
      361 tagtcgatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
      421 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
      481 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
      541 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
      601 ataataatgg tttcttagga cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt
      661 taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt tttgtatttg
      721 gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa aaaaataaac
      781 aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt agacaagaaa
      841 agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat cacaggattt
      901 tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac ctgagagcag
      961 gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt tacatcttcg
     1021 gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt aatttatata
     1081 tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa aggacccagg
     1141 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1201 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1261 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1321 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1381 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1441 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1501 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1561 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1621 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1681 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1741 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1801 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1861 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1921 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1981 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2041 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2101 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2161 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2221 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2281 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2341 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2401 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2461 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2521 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2581 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2641 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2701 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2761 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2821 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2881 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2941 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3001 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3061 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3121 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     3181 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     3241 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     3301 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctccgga tgcaagggtt
     3361 cgaatccctt agctctcatt attttttgct ttttctcttg aggtcacatg atcgcaaaat
     3421 ggcaaatggc acgtgaagct gtcgatattg gggaactgtg gtggttggca aatgactaat
     3481 taagttagtc aaggcgccat cctcatgaaa actgtgtaac ataataaccg aagtgtcgaa
     3541 aaggtggcac cttgtccaat tgaacacgct cgatgaaaaa aataagatat atataaggtt
     3601 aagtaaagcg tctgttagaa aggaagtttt tcctttttct tgctctcttg tcttttcatc
     3661 tactatttcc ttcgtgtaat acagggtcgt cagatacata gatacaattc tattaccccc
     3721 atccatactc tagaactagt ggatccatgg tggttttggc tgcttctatc actacgcgtc
     3781 aggggaaacc tctcttgtcc aggcaattca aagatctgtc caaggatcga gtcctagagt
     3841 tgctgtccaa tttccagaac ctagtctccg agatctcgtc agaccacaca tttgtcgagg
     3901 acaagcacgt ccgctacgtg tataggccct ttgataacta ctacattatt cttatcacaa
     3961 accgccagtc caatattatc aaggatttgg ccacattaaa tctgttttca cagacgatca
     4021 actcctactt gtctagtttc caagaccaag agatttttca caatgccttt gagatcttgt
     4081 cctcgttcga cgaaatcgtg tcaatgggcg ggtacaaaga gaacttgtcc tttactcaag
     4141 tgcaaacata cttgtctatg gagtcgcacg aagaaagaat cttcgaaatt attgagcgta
     4201 ataaagagat cgaagctact gaggagagga agcgccgtgc taaggaaatt gctagaaaag
     4261 aacatgaaag gaaacacggg catcaccatc accatcacta gccgctcgag tcatgtaatt
     4321 agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg
     4381 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
     4441 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
     4501 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg
     4561 ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     4621 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     4681 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     4741 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4801 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4861 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4921 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4981 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5041 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5101 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5161 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
     5221 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat cgaccggtcg
     5281 aggagaactt ctagtatatc tacataccta atattattgc cttattaaaa atggaatccc
     5341 aacaattaca tcaaaatcca cattctcttc aaaatcaatt gtcctgtact tccttgttca
     5401 tgtgtgttca aaaacgttat atttatagga taattatact ctatttctca acaagtaatt
     5461 ggttgtttgg ccgagcggtc taaggcgcct gattcaagaa atatcttgac cgcagttaac
     5521 tgtgggaata ctcaggtatc gtaagatgca agagttcgaa tctcttagca accattattt
     5581 ttttcctcaa cataacgaga acacacaggg gcgctatcgc acagaatcaa attcgatgac
     5641 tggaaatttt ttgttaattt cagaggtcgc ctgacgcata tacctttttc aactgaaaaa
     5701 ttgggagaaa aaggaaaggt gagagcgccg gaaccggctt ttcatataga atagagaagc
     5761 gttcatgact aaatgcttgc atcacaatac ttgaagttga caatattatt taaggaccta
     5821 ttgttttttc caataggtgg ttagcaatcg tcttactttc taacttttct taccttttac
     5881 atttcagcaa tatatatata tatatttcaa ggatatacca ttctaatgtc tgcccctaag
     5941 aagatcgtcg ttttgccagg tgaccacgtt ggtcaagaaa tcacagccga agccattaag
     6001 gttcttaaag ctatttctga tgttcgttcc aatgtcaagt tcgatttcga aaatcattta
     6061 attggtggtg ctgctatcga tgctacaggt gttccacttc cagatgaggc gctggaagcc
     6121 tccaagaagg ctgatgccgt tttgttaggt gctgtgggtg gtcctaaatg gggtaccggt
     6181 agtgttagac ctgaacaagg tttactaaaa atccgtaaag aacttcaatt gtacgccaac
     6241 ttaagaccat gtaactttgc atccgactct cttttagact tatctccaat caagccacaa
     6301 tttgctaaag gtactgactt cgttgttgtc agagaattag tgggaggtat ttactttggt
     6361 aagagaaagg aagacgatgg tgatggtgtc gcttgggata gtgaacaata caccgttcca
     6421 gaagtgcaaa gaatcacaag aatggccgct ttcatggccc tacaacatga gccaccattg
     6481 cctatttggt ccttggataa agctaatgtt ttggcctctt caagattatg gagaaaaact
     6541 gtggaggaaa ccatcaagaa cgaattccct acattgaagg ttcaacatca attgattgat
     6601 tctgccgcca tgatcctagt taagaaccca acccacctaa atggtattat aatcaccagc
     6661 aacatgtttg gtgatatcat ctccgatgaa gcctccgtta tcccaggttc cttgggtttg
     6721 ttgccatctg cgtccttggc ctctttgcca gacaagaaca ccgcatttgg tttgtacgaa
     6781 ccatgccacg gttctgctcc agatttgcca aagaataagg tcaaccctat cgccactatc
     6841 ttgtctgctg caatgatgtt gaaattgtca ttgaacttgc ctgaagaagg taaggccatt
     6901 gaagatgcag ttaaaaaggt tttggatgca ggtatcagaa ctggtgattt aggtggttcc
     6961 aacagtacca ccgaagtcgg tgatgctgtc gccgaagaag ttaagaaaat ccttgcttaa
     7021 aaagattctc tttttttatg atatttgtac ataaacttta taaatgaaat tcataataga
     7081 aacgacacga aattacaaaa tggaatatgt tcatagggta gacgaaacta
//
