LOCUS       sequence-200051-        7163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chimaera 12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7163)
  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 7163)
  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-12
FEATURES             Location/Qualifiers
     source          1..7163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(81..100)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     200..222
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(260..278)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    320..823
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        860..964
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(965..1033,1034..1825)
                     /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(1183..1202)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1996..2584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2485..2504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2738..2755
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2872..2893
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2908..2913,2914..2931,2932..2938)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2946..2962
                     /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     2951..2973
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2970..2986
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2970..2986
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3005..3025
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3007..3025
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3076..3425
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3426..3442
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3426..3442
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3978..3995
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4022..4038)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4023..4039)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4041..4288
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4057..4075)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4306..4325)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4307..4325)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4335..4352)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4335..4351)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4344..4366)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4492..4947
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4579..4598)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4789..4810
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5248..5655
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5656..6750
                     /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 cataatggtg aaagttccct caagaatttt actctgtcag aaacggcctt aacgacgtag
       61 tcgatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
      121 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
      181 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
      241 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
      301 ataatggttt cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa
      361 tgatggaata atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat
      421 tttagaaagt aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa
      481 ggtttaaaaa atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc
      541 agattaaata gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg
      601 tgtgtggtct tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa
      661 gagcaagata aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa
      721 aacaaaaact attttttctt taatttcttt ttttactttc tatttttaat ttatatattt
      781 atattaaaaa atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg
      841 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      901 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      961 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1021 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     1081 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1141 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1201 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1261 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1321 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1381 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1441 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1501 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1561 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1621 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1681 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1741 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1801 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1861 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1921 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1981 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2041 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2101 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2161 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2221 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2281 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2341 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2401 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2461 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2521 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2581 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2641 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2701 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2761 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2821 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     2881 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     2941 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     3001 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctccggatgc aagggttcga
     3061 atcccttagc tctcattatt ttttgctttt tctcttgagg tcacatgatc gcaaaatggc
     3121 aaatggcacg tgaagctgtc gatattgggg aactgtggtg gttggcaaat gactaattaa
     3181 gttagtcaag gcgccatcct catgaaaact gtgtaacata ataaccgaag tgtcgaaaag
     3241 gtggcacctt gtccaattga acacgctcga tgaaaaaaat aagatatata taaggttaag
     3301 taaagcgtct gttagaaagg aagtttttcc tttttcttgc tctcttgtct tttcatctac
     3361 tatttccttc gtgtaataca gggtcgtcag atacatagat acaattctat tacccccatc
     3421 catactctag aactagtgga tccaccatgg tgctgttggc agcagcagtc tgtacaaaag
     3481 caggaaaagc tattgtttct cgacagtttg tagagatgac tcgaactcgg attgagggct
     3541 tgttagcagc ttttccaaag ctcatgaaca ctggaaaaca acatacattt gtggaaacag
     3601 agagtgtcag atatgtttac cagcctatgg agaaattgta catggtactg atcaccaccc
     3661 gccagtccaa tattatcaag gatttggcca cattaaatct gttttcacag acgatcaact
     3721 cctacttgtc tagtttccaa gaccaagaga tttttcacaa tgcctttgag atcttgtcct
     3781 cgttcgacga aatcgtgtca atgggcgggt acaaagagaa cttgtccttt actcaagtgc
     3841 aaacatactt gtctatggag tcgcacgaag aaagaatcca ggaaattatt gagcgtaata
     3901 aagagatcga agctactgag gagaggaagc gccgtgctaa ggaaattgct agaaaagaac
     3961 atgaaaggaa acacgggcat caccatcacc atcactgacc ggaattcgat atcaagctta
     4021 tcgataccgt cgacctcgag tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     4081 ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     4141 ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt
     4201 ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt
     4261 tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta tagtgagtcg
     4321 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     4381 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     4441 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     4501 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4561 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4621 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4681 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4741 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4801 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4861 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4921 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     4981 cggtatttca caccgcatat cgaccggtcg aggagaactt ctagtatatc tacataccta
     5041 atattattgc cttattaaaa atggaatccc aacaattaca tcaaaatcca cattctcttc
     5101 aaaatcaatt gtcctgtact tccttgttca tgtgtgttca aaaacgttat atttatagga
     5161 taattatact ctatttctca acaagtaatt ggttgtttgg ccgagcggtc taaggcgcct
     5221 gattcaagaa atatcttgac cgcagttaac tgtgggaata ctcaggtatc gtaagatgca
     5281 agagttcgaa tctcttagca accattattt ttttcctcaa cataacgaga acacacaggg
     5341 gcgctatcgc acagaatcaa attcgatgac tggaaatttt ttgttaattt cagaggtcgc
     5401 ctgacgcata tacctttttc aactgaaaaa ttgggagaaa aaggaaaggt gagagcgccg
     5461 gaaccggctt ttcatataga atagagaagc gttcatgact aaatgcttgc atcacaatac
     5521 ttgaagttga caatattatt taaggaccta ttgttttttc caataggtgg ttagcaatcg
     5581 tcttactttc taacttttct taccttttac atttcagcaa tatatatata tatatttcaa
     5641 ggatatacca ttctaatgtc tgcccctaag aagatcgtcg ttttgccagg tgaccacgtt
     5701 ggtcaagaaa tcacagccga agccattaag gttcttaaag ctatttctga tgttcgttcc
     5761 aatgtcaagt tcgatttcga aaatcattta attggtggtg ctgctatcga tgctacaggt
     5821 gttccacttc cagatgaggc gctggaagcc tccaagaagg ctgatgccgt tttgttaggt
     5881 gctgtgggtg gtcctaaatg gggtaccggt agtgttagac ctgaacaagg tttactaaaa
     5941 atccgtaaag aacttcaatt gtacgccaac ttaagaccat gtaactttgc atccgactct
     6001 cttttagact tatctccaat caagccacaa tttgctaaag gtactgactt cgttgttgtc
     6061 agagaattag tgggaggtat ttactttggt aagagaaagg aagacgatgg tgatggtgtc
     6121 gcttgggata gtgaacaata caccgttcca gaagtgcaaa gaatcacaag aatggccgct
     6181 ttcatggccc tacaacatga gccaccattg cctatttggt ccttggataa agctaatgtt
     6241 ttggcctctt caagattatg gagaaaaact gtggaggaaa ccatcaagaa cgaattccct
     6301 acattgaagg ttcaacatca attgattgat tctgccgcca tgatcctagt taagaaccca
     6361 acccacctaa atggtattat aatcaccagc aacatgtttg gtgatatcat ctccgatgaa
     6421 gcctccgtta tcccaggttc cttgggtttg ttgccatctg cgtccttggc ctctttgcca
     6481 gacaagaaca ccgcatttgg tttgtacgaa ccatgccacg gttctgctcc agatttgcca
     6541 aagaataagg tcaaccctat cgccactatc ttgtctgctg caatgatgtt gaaattgtca
     6601 ttgaacttgc ctgaagaagg taaggccatt gaagatgcag ttaaaaaggt tttggatgca
     6661 ggtatcagaa ctggtgattt aggtggttcc aacagtacca ccgaagtcgg tgatgctgtc
     6721 gccgaagaag ttaagaaaat ccttgcttaa aaagattctc tttttttatg atatttgtac
     6781 ataaacttta taaatgaaat tcataataga aacgacacga aattacaaaa tggaatatgt
     6841 tcatagggta gacgaaacta tatacgcaat ctacatacat ttatcaagaa ggagaaaaag
     6901 gaggatgtaa aggaatacag gtaagcaaat tgatactaat ggctcaacgt gataaggaaa
     6961 aagaattgca ctttaacatt aatattgaca aggaggaggg caccacacaa aaagttaggt
     7021 gtaacagaaa atcatgaaac tatgattcct aatttatata ttggaggatt ttctctaaaa
     7081 aaaaaaaaat acaacaaata aaaaacactc aatgacctga ccatttgatg gagtttaagt
     7141 caataccttc ttgaaccatt tcc
//
