LOCUS       sequence-198858-        8262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ret2 (full length).
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
FEATURES             Location/Qualifiers
     source          1..8262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /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(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2622..2643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /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     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2755..2775
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2826..3175
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3176..3192
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3176..3192
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4871..4887)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4872..4888)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4890..5137
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4906..4924)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5155..5174)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5156..5174)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5184..5201)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5184..5200)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5193..5215)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5341..5796
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5428..5447)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5638..5659
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6097..6504
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6505..7599
                     /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"
     primer_bind     complement(8093..8112)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8212..8234
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2761 aaccctcact aaagggaaca aaagctggag ctccggatgc aagggttcga atcccttagc
     2821 tctcattatt ttttgctttt tctcttgagg tcacatgatc gcaaaatggc aaatggcacg
     2881 tgaagctgtc gatattgggg aactgtggtg gttggcaaat gactaattaa gttagtcaag
     2941 gcgccatcct catgaaaact gtgtaacata ataaccgaag tgtcgaaaag gtggcacctt
     3001 gtccaattga acacgctcga tgaaaaaaat aagatatata taaggttaag taaagcgtct
     3061 gttagaaagg aagtttttcc tttttcttgc tctcttgtct tttcatctac tatttccttc
     3121 gtgtaataca gggtcgtcag atacatagat acaattctat tacccccatc catactctag
     3181 aactagtgga tccatggtgg ttttggctgc ttctatcact acgcgtcagg ggaaacctct
     3241 cttgtccagg caattcaaag atctgtccaa ggatcgagtc ctagagttgc tgtccaattt
     3301 ccagaaccta gtctccgaga tctcgtcaga ccacacattt gtcgaggaca agcacgtccg
     3361 ctacgtgtat aggccctttg ataactacta cattattctt atcacaaacc gccagtccaa
     3421 tattatcaag gatttggcca cattaaatct gttttcacag acgatcaact cctacttgtc
     3481 tagtttccaa gaccaagaga tttttcacaa tgcctttgag atcttgtcct cgttcgacga
     3541 aatcgtgtca atgggcgggt acaaagagaa cttgtccttt actcaagtgc aaacatactt
     3601 gtctatggag tcgcacgaag aaagaatcca ggaaattatt gagcgtaata aagagatcga
     3661 agctactgag gagaggaagc gccgtgctaa ggaaattgct agaaaagaac atgaaaggaa
     3721 acacgggttt atgtcttcaa atggcgatta cgacggcgcc aataggttca tgggaagtaa
     3781 agatcctaac gttacaaacg ccatcaacag ttactactcc catgcttctc ctgcagcaca
     3841 acagtcttat ctacaaagtt cccatgccgc tgctgctgag gtggccccag tggcctcccc
     3901 aatggctacg tctcagcgtg cgggacattc tgctactggc ggtatgaaat tgggtggtgg
     3961 tgccggaaga agagcgggtg ccgcaccacg gccaagtgca atttcttcag cctcttctgg
     4021 cactccacct cctccggagg aggatgtccc agagaacaat ggtattttga tctctatcaa
     4081 ggaggtaatc aatgcagaat tctccagaga tggtaccatt cactcttctg aattgaaagg
     4141 tgttttagaa ctaagaatta atgaccatga cttatctcac tcgaacctaa aactagctga
     4201 ttctattgac gttcgcgata aatccttcca gtttaagacc catccaaaca tagataaaca
     4261 atcgttccta tccactaaac ttatctccct gcgtgataaa tctaaggctt tcccagccaa
     4321 cgaccaaagc ctgggtgttc taagatggcg taaagtcgct cccgcagaag atgactcatt
     4381 aattccatta actctaacta catgggtttc cccctccgaa tcacaacagg gctttgatgt
     4441 cattattgag tatgagagcg tcctggaaac agaactcgct gatgtgatct tcaccattcc
     4501 agttttccct caggagccag tagacataaa tacagagtct tctacatgct ctgacgctga
     4561 agttgtcaac atggaccaag aaatgggaac ctccattaag attagcaaaa ttgctgccaa
     4621 cgatgcaggt gcgctagcat tcactatcga agctccatat gaagatgcat tgtacccaat
     4681 gaccgtttcc ttccaagagt ccacgaggga taaattagct aagagtttca ccggtatggc
     4741 catacaatct gttgtcatgg ccaatgacca cgatcaagag ctcccttacg acgtcatcac
     4801 ctctttgaaa tcagatgaat atcttgtcca atagtccccc cgggctgcag gaattcgata
     4861 tcaagcttat cgataccgtc gacctcgagt catgtaatta gttatgtcac gcttacattc
     4921 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     4981 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     5041 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     5101 tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc cggtacccaa ttcgccctat
     5161 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     5221 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     5281 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     5341 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     5401 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     5461 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     5521 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     5581 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     5641 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     5701 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     5761 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     5821 gcatctgtgc ggtatttcac accgcatatc gaccggtcga ggagaacttc tagtatatct
     5881 acatacctaa tattattgcc ttattaaaaa tggaatccca acaattacat caaaatccac
     5941 attctcttca aaatcaattg tcctgtactt ccttgttcat gtgtgttcaa aaacgttata
     6001 tttataggat aattatactc tatttctcaa caagtaattg gttgtttggc cgagcggtct
     6061 aaggcgcctg attcaagaaa tatcttgacc gcagttaact gtgggaatac tcaggtatcg
     6121 taagatgcaa gagttcgaat ctcttagcaa ccattatttt tttcctcaac ataacgagaa
     6181 cacacagggg cgctatcgca cagaatcaaa ttcgatgact ggaaattttt tgttaatttc
     6241 agaggtcgcc tgacgcatat acctttttca actgaaaaat tgggagaaaa aggaaaggtg
     6301 agagcgccgg aaccggcttt tcatatagaa tagagaagcg ttcatgacta aatgcttgca
     6361 tcacaatact tgaagttgac aatattattt aaggacctat tgttttttcc aataggtggt
     6421 tagcaatcgt cttactttct aacttttctt accttttaca tttcagcaat atatatatat
     6481 atatttcaag gatataccat tctaatgtct gcccctaaga agatcgtcgt tttgccaggt
     6541 gaccacgttg gtcaagaaat cacagccgaa gccattaagg ttcttaaagc tatttctgat
     6601 gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa ttggtggtgc tgctatcgat
     6661 gctacaggtg ttccacttcc agatgaggcg ctggaagcct ccaagaaggc tgatgccgtt
     6721 ttgttaggtg ctgtgggtgg tcctaaatgg ggtaccggta gtgttagacc tgaacaaggt
     6781 ttactaaaaa tccgtaaaga acttcaattg tacgccaact taagaccatg taactttgca
     6841 tccgactctc ttttagactt atctccaatc aagccacaat ttgctaaagg tactgacttc
     6901 gttgttgtca gagaattagt gggaggtatt tactttggta agagaaagga agacgatggt
     6961 gatggtgtcg cttgggatag tgaacaatac accgttccag aagtgcaaag aatcacaaga
     7021 atggccgctt tcatggccct acaacatgag ccaccattgc ctatttggtc cttggataaa
     7081 gctaatgttt tggcctcttc aagattatgg agaaaaactg tggaggaaac catcaagaac
     7141 gaattcccta cattgaaggt tcaacatcaa ttgattgatt ctgccgccat gatcctagtt
     7201 aagaacccaa cccacctaaa tggtattata atcaccagca acatgtttgg tgatatcatc
     7261 tccgatgaag cctccgttat cccaggttcc ttgggtttgt tgccatctgc gtccttggcc
     7321 tctttgccag acaagaacac cgcatttggt ttgtacgaac catgccacgg ttctgctcca
     7381 gatttgccaa agaataaggt caaccctatc gccactatct tgtctgctgc aatgatgttg
     7441 aaattgtcat tgaacttgcc tgaagaaggt aaggccattg aagatgcagt taaaaaggtt
     7501 ttggatgcag gtatcagaac tggtgattta ggtggttcca acagtaccac cgaagtcggt
     7561 gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa aagattctct ttttttatga
     7621 tatttgtaca taaactttat aaatgaaatt cataatagaa acgacacgaa attacaaaat
     7681 ggaatatgtt catagggtag acgaaactat atacgcaatc tacatacatt tatcaagaag
     7741 gagaaaaagg aggatgtaaa ggaatacagg taagcaaatt gatactaatg gctcaacgtg
     7801 ataaggaaaa agaattgcac tttaacatta atattgacaa ggaggagggc accacacaaa
     7861 aagttaggtg taacagaaaa tcatgaaact atgattccta atttatatat tggaggattt
     7921 tctctaaaaa aaaaaaaata caacaaataa aaaacactca atgacctgac catttgatgg
     7981 agtttaagtc aataccttct tgaaccattt cccataatgg tgaaagttcc ctcaagaatt
     8041 ttactctgtc agaaacggcc ttaacgacgt agtcgatatg gtgcactctc agtacaatct
     8101 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     8161 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     8221 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc ga
//
