LOCUS       sequence-199958-        8187 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chimaera 1 (Ret2-COPD).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8187)
  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 8187)
  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 CHI-1
FEATURES             Location/Qualifiers
     source          1..8187
                     /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(4796..4812)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4797..4813)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4815..5062
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4831..4849)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5080..5099)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5081..5099)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5109..5126)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5109..5125)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5118..5140)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5266..5721
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5353..5372)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5563..5584
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6022..6429
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6430..7524
                     /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(8018..8037)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8137..8159
                     /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 cagcctatgg agaaattgta catggtactg atcaccacca aaaacagcaa
     3421 catcttagaa gatctggaga ccctaaggct cttctcaaga gtgattcctg aatattgccg
     3481 agccttagaa gagaatgaaa tatctgagca ctgttttgat ttgatttttg catttgacga
     3541 aattgttgcc ttgggatacc gggagaatgt taacctggca cagatcagaa ccttcacaga
     3601 aatggattcc catgaggaga aggtgttcag agcagtcaga gagactcaag agcgtgaagc
     3661 caaggctgag atgcggcgta aagcaaaaga attacagcag gcccgaagag atgcagagag
     3721 acagggcaaa aaagcaccag gatttggggg ctttggaagc tccacagtgt ctggaggcag
     3781 cacaacttct atgatcacag agaccatcat tgaaactgac aaaccaaaag tggcacccgc
     3841 accagccaga ccttcaggcc ctagcaaggc tctgaaactg ggagccaaag gaaaggaagt
     3901 agataacttc gttgacaaat tgaaatctga aggcgaaaat atcatatcct ccaatatggg
     3961 caagcgtacc tctgaagcaa ccaaagtgca tgctccaccc attaatatgg agagtgtgca
     4021 catgaagatt gaggaaaaga tcacattaac ctgtggccga gatggaggat tacagaacat
     4081 ggagctgcac ggcatgatca tgcttaggat ctcagatgat aaatttggcc gaattcgtct
     4141 tcatgtggaa aatgaagaca agaaaggagt acagctacag acccatccaa atgtggataa
     4201 aaaacttttc actgcagaat ctctaattgg tttgaagaat ccagagaagt catttccagt
     4261 caacagtgat gtaggggtgc taaagtggag actgcagacc acagaggagt cttttattcc
     4321 actgacaatt aattgctggc cctcagaaag tggaaatggt tgtgatgtca acatagaata
     4381 tgagctacaa gaagataatt tagaactgaa tgatgtggtt atcaccattc cactcccatc
     4441 tggtgttggc gctccagtga ttggtgagat tgatggcgag tatcgacatg acagtcgacg
     4501 aaataccttg gagtggtgcc tgccagtgat tgatgccaaa aataagagtg gcagccttga
     4561 gttcagcatt gctgggcagc ccaatgactt cttccctgtt caagtctcct tcatctccaa
     4621 aaaaaattac tgtaacatac aggttaccaa agtgacccag gtagatggaa acagccctgt
     4681 aaggttttcc acagagacca ctttcctagt ggataagtat gaaattctgt agataagaat
     4741 gcggccgctc tagaactagt gcccccgggc tgcaggaatt cgatatcaag cttatcgata
     4801 ccgtcgacct cgagtcatgt aattagttat gtcacgctta cattcacgcc ctccccccac
     4861 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
     4921 ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
     4981 gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg
     5041 acgctcgaag gctttaattt gcggccggta cccaattcgc cctatagtga gtcgtattac
     5101 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     5161 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     5221 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg
     5281 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5341 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5401 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     5461 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5521 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5581 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     5641 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5701 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     5761 ttcacaccgc atatcgaccg gtcgaggaga acttctagta tatctacata cctaatatta
     5821 ttgccttatt aaaaatggaa tcccaacaat tacatcaaaa tccacattct cttcaaaatc
     5881 aattgtcctg tacttccttg ttcatgtgtg ttcaaaaacg ttatatttat aggataatta
     5941 tactctattt ctcaacaagt aattggttgt ttggccgagc ggtctaaggc gcctgattca
     6001 agaaatatct tgaccgcagt taactgtggg aatactcagg tatcgtaaga tgcaagagtt
     6061 cgaatctctt agcaaccatt atttttttcc tcaacataac gagaacacac aggggcgcta
     6121 tcgcacagaa tcaaattcga tgactggaaa ttttttgtta atttcagagg tcgcctgacg
     6181 catatacctt tttcaactga aaaattggga gaaaaaggaa aggtgagagc gccggaaccg
     6241 gcttttcata tagaatagag aagcgttcat gactaaatgc ttgcatcaca atacttgaag
     6301 ttgacaatat tatttaagga cctattgttt tttccaatag gtggttagca atcgtcttac
     6361 tttctaactt ttcttacctt ttacatttca gcaatatata tatatatatt tcaaggatat
     6421 accattctaa tgtctgcccc taagaagatc gtcgttttgc caggtgacca cgttggtcaa
     6481 gaaatcacag ccgaagccat taaggttctt aaagctattt ctgatgttcg ttccaatgtc
     6541 aagttcgatt tcgaaaatca tttaattggt ggtgctgcta tcgatgctac aggtgttcca
     6601 cttccagatg aggcgctgga agcctccaag aaggctgatg ccgttttgtt aggtgctgtg
     6661 ggtggtccta aatggggtac cggtagtgtt agacctgaac aaggtttact aaaaatccgt
     6721 aaagaacttc aattgtacgc caacttaaga ccatgtaact ttgcatccga ctctctttta
     6781 gacttatctc caatcaagcc acaatttgct aaaggtactg acttcgttgt tgtcagagaa
     6841 ttagtgggag gtatttactt tggtaagaga aaggaagacg atggtgatgg tgtcgcttgg
     6901 gatagtgaac aatacaccgt tccagaagtg caaagaatca caagaatggc cgctttcatg
     6961 gccctacaac atgagccacc attgcctatt tggtccttgg ataaagctaa tgttttggcc
     7021 tcttcaagat tatggagaaa aactgtggag gaaaccatca agaacgaatt ccctacattg
     7081 aaggttcaac atcaattgat tgattctgcc gccatgatcc tagttaagaa cccaacccac
     7141 ctaaatggta ttataatcac cagcaacatg tttggtgata tcatctccga tgaagcctcc
     7201 gttatcccag gttccttggg tttgttgcca tctgcgtcct tggcctcttt gccagacaag
     7261 aacaccgcat ttggtttgta cgaaccatgc cacggttctg ctccagattt gccaaagaat
     7321 aaggtcaacc ctatcgccac tatcttgtct gctgcaatga tgttgaaatt gtcattgaac
     7381 ttgcctgaag aaggtaaggc cattgaagat gcagttaaaa aggttttgga tgcaggtatc
     7441 agaactggtg atttaggtgg ttccaacagt accaccgaag tcggtgatgc tgtcgccgaa
     7501 gaagttaaga aaatccttgc ttaaaaagat tctctttttt tatgatattt gtacataaac
     7561 tttataaatg aaattcataa tagaaacgac acgaaattac aaaatggaat atgttcatag
     7621 ggtagacgaa actatatacg caatctacat acatttatca agaaggagaa aaaggaggat
     7681 gtaaaggaat acaggtaagc aaattgatac taatggctca acgtgataag gaaaaagaat
     7741 tgcactttaa cattaatatt gacaaggagg agggcaccac acaaaaagtt aggtgtaaca
     7801 gaaaatcatg aaactatgat tcctaattta tatattggag gattttctct aaaaaaaaaa
     7861 aaatacaaca aataaaaaac actcaatgac ctgaccattt gatggagttt aagtcaatac
     7921 cttcttgaac catttcccat aatggtgaaa gttccctcaa gaattttact ctgtcagaaa
     7981 cggccttaac gacgtagtcg atatggtgca ctctcagtac aatctgctct gatgccgcat
     8041 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     8101 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     8161 tttcaccgtc atcaccgaaa cgcgcga
//
