LOCUS       sequence-200181-        7130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N152T 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 N152T
FEATURES             Location/Qualifiers
     source          1..7130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(190..209)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     309..331
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(369..387)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    429..932
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        969..1073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1074..1142,1143..1934)
                     /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(1292..1311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2105..2693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2594..2613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2847..2864
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2981..3002
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3017..3022,3023..3040,3041..3047)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3055..3071
                     /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     3060..3082
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3079..3095
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3079..3095
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3114..3134
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3116..3134
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3185..3534
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3535..3551
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3535..3551
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4087..4104
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4117..4364
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4133..4151)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4382..4401)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4383..4401)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4411..4428)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4411..4427)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4420..4442)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4568..5023
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4655..4674)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4865..4886
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5324..5731
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5732..6826
                     /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 tatatattgg aggattttct ctaaaaaaaa aaaaatacaa caaataaaaa acactcaatg
       61 acctgaccat ttgatggagt ttaagtcaat accttcttga accatttccc ataatggtga
      121 aagttccctc aagaatttta ctctgtcaga aacggcctta acgacgtagt cgatatggtg
      181 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
      241 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
      301 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
      361 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
      421 ttaggacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa
      481 tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta
      541 aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa
      601 tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag
      661 atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt
      721 ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa
      781 aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta
      841 ttttttcttt aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa
      901 tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg
      961 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1021 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1081 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1141 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1201 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1261 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1321 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1381 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1441 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1501 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1561 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1621 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1681 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1741 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1801 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1861 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1921 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1981 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2041 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2101 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2161 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2221 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2281 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2341 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2401 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2461 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2521 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2581 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2641 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2701 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2761 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2821 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2881 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     2941 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     3001 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     3061 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     3121 accctcacta aagggaacaa aagctggagc tccggatgca agggttcgaa tcccttagct
     3181 ctcattattt tttgcttttt ctcttgaggt cacatgatcg caaaatggca aatggcacgt
     3241 gaagctgtcg atattgggga actgtggtgg ttggcaaatg actaattaag ttagtcaagg
     3301 cgccatcctc atgaaaactg tgtaacataa taaccgaagt gtcgaaaagg tggcaccttg
     3361 tccaattgaa cacgctcgat gaaaaaaata agatatatat aaggttaagt aaagcgtctg
     3421 ttagaaagga agtttttcct ttttcttgct ctcttgtctt ttcatctact atttccttcg
     3481 tgtaatacag ggtcgtcaga tacatagata caattctatt acccccatcc atactctaga
     3541 actagtggat ccatggtggt tttggctgct tctatcacta cgcgtcaggg gaaacctctc
     3601 ttgtccaggc aattcaaaga tctgtccaag gatcgagtcc tagagttgct gtccaatttc
     3661 cagaacctag tctccgagat ctcgtcagac cacacatttg tcgaggacaa gcacgtccgc
     3721 tacgtgtata ggccctttga taactactac attattctta tcacaaaccg ccagtccaat
     3781 attatcaagg atttggccac attaaatctg ttttcacaga cgatcaactc ctacttgtct
     3841 agtttccaag accaagagat ttttcacaat gcctttgaga tcttgtcctc gttcgacgaa
     3901 atcgtgtcaa tgggcgggta caaagagaac ttgtccttta ctcaagtgca aacatacttg
     3961 tctatggagt cgcacgaaga aagaatccag gaaattattg agcgtactaa agagatcgaa
     4021 gctactgagg agaggaagcg ccgtgctaag gaaattgcta gaaaagaaca tgaaaggaaa
     4081 cacgggcatc accatcacca tcactagccg ctcgagtcat gtaattagtt atgtcacgct
     4141 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     4201 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     4261 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     4321 ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcggccgg tacccaattc
     4381 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     4441 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     4501 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4561 cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4621 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4681 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4741 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4801 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4861 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4921 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4981 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     5041 tccttacgca tctgtgcggt atttcacacc gcatatcgac cggtcgagga gaacttctag
     5101 tatatctaca tacctaatat tattgcctta ttaaaaatgg aatcccaaca attacatcaa
     5161 aatccacatt ctcttcaaaa tcaattgtcc tgtacttcct tgttcatgtg tgttcaaaaa
     5221 cgttatattt ataggataat tatactctat ttctcaacaa gtaattggtt gtttggccga
     5281 gcggtctaag gcgcctgatt caagaaatat cttgaccgca gttaactgtg ggaatactca
     5341 ggtatcgtaa gatgcaagag ttcgaatctc ttagcaacca ttattttttt cctcaacata
     5401 acgagaacac acaggggcgc tatcgcacag aatcaaattc gatgactgga aattttttgt
     5461 taatttcaga ggtcgcctga cgcatatacc tttttcaact gaaaaattgg gagaaaaagg
     5521 aaaggtgaga gcgccggaac cggcttttca tatagaatag agaagcgttc atgactaaat
     5581 gcttgcatca caatacttga agttgacaat attatttaag gacctattgt tttttccaat
     5641 aggtggttag caatcgtctt actttctaac ttttcttacc ttttacattt cagcaatata
     5701 tatatatata tttcaaggat ataccattct aatgtctgcc cctaagaaga tcgtcgtttt
     5761 gccaggtgac cacgttggtc aagaaatcac agccgaagcc attaaggttc ttaaagctat
     5821 ttctgatgtt cgttccaatg tcaagttcga tttcgaaaat catttaattg gtggtgctgc
     5881 tatcgatgct acaggtgttc cacttccaga tgaggcgctg gaagcctcca agaaggctga
     5941 tgccgttttg ttaggtgctg tgggtggtcc taaatggggt accggtagtg ttagacctga
     6001 acaaggttta ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa gaccatgtaa
     6061 ctttgcatcc gactctcttt tagacttatc tccaatcaag ccacaatttg ctaaaggtac
     6121 tgacttcgtt gttgtcagag aattagtggg aggtatttac tttggtaaga gaaaggaaga
     6181 cgatggtgat ggtgtcgctt gggatagtga acaatacacc gttccagaag tgcaaagaat
     6241 cacaagaatg gccgctttca tggccctaca acatgagcca ccattgccta tttggtcctt
     6301 ggataaagct aatgttttgg cctcttcaag attatggaga aaaactgtgg aggaaaccat
     6361 caagaacgaa ttccctacat tgaaggttca acatcaattg attgattctg ccgccatgat
     6421 cctagttaag aacccaaccc acctaaatgg tattataatc accagcaaca tgtttggtga
     6481 tatcatctcc gatgaagcct ccgttatccc aggttccttg ggtttgttgc catctgcgtc
     6541 cttggcctct ttgccagaca agaacaccgc atttggtttg tacgaaccat gccacggttc
     6601 tgctccagat ttgccaaaga ataaggtcaa ccctatcgcc actatcttgt ctgctgcaat
     6661 gatgttgaaa ttgtcattga acttgcctga agaaggtaag gccattgaag atgcagttaa
     6721 aaaggttttg gatgcaggta tcagaactgg tgatttaggt ggttccaaca gtaccaccga
     6781 agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag attctctttt
     6841 tttatgatat ttgtacataa actttataaa tgaaattcat aatagaaacg acacgaaatt
     6901 acaaaatgga atatgttcat agggtagacg aaactatata cgcaatctac atacatttat
     6961 caagaaggag aaaaaggagg atgtaaagga atacaggtaa gcaaattgat actaatggct
     7021 caacgtgata aggaaaaaga attgcacttt aacattaata ttgacaagga ggagggcacc
     7081 acacaaaaag ttaggtgtaa cagaaaatca tgaaactatg attcctaatt
//
