LOCUS       sequence-200054-        7142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chimaera 15.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7142)
  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 7142)
  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-15
FEATURES             Location/Qualifiers
     source          1..7142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(185..204)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     304..326
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(364..382)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    424..927
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        964..1068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1069..1137,1138..1929)
                     /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(1287..1306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2100..2688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2589..2608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2842..2859
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2976..2997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3012..3017,3018..3035,3036..3042)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3050..3066
                     /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     3055..3077
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3074..3090
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3074..3090
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3109..3129
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3111..3129
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3180..3529
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3530..3546
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3530..3546
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4082..4099
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4124..4371
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4140..4158)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4389..4408)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4390..4408)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4418..4435)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4418..4434)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4427..4449)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4575..5030
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4662..4681)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4872..4893
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5331..5738
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5739..6833
                     /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 attggaggat tttctctaaa aaaaaaaaaa tacaacaaat aaaaaacact caatgacctg
       61 accatttgat ggagtttaag tcaatacctt cttgaaccat ttcccataat ggtgaaagtt
      121 ccctcaagaa ttttactctg tcagaaacgg ccttaacgac gtagtcgata tggtgcactc
      181 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
      241 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
      301 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
      361 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttagg
      421 acggatcgct tgcctgtaac ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg
      481 gaatttactc tgtgtttatt tatttttatg ttttgtattt ggattttaga aagtaaataa
      541 agaaggtaga agagttacgg aatgaagaaa aaaaaataaa caaaggttta aaaaatttca
      601 acaaaaagcg tactttacat atatatttat tagacaagaa aagcagatta aatagatata
      661 cattcgatta acgataagta aaatgtaaaa tcacaggatt ttcgtgtgtg gtcttctaca
      721 cagacaagat gaaacaattc ggcattaata cctgagagca ggaagagcaa gataaaaggt
      781 agtatttgtt ggcgatcccc ctagagtctt ttacatcttc ggaaaacaaa aactattttt
      841 tctttaattt ctttttttac tttctatttt taatttatat atttatatta aaaaatttaa
      901 attataatta tttttatagc acgtgatgaa aaggacccag gtggcacttt tcggggaaat
      961 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1021 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     1081 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     1141 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1201 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1261 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1321 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1381 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1441 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1501 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1561 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1621 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1681 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1741 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     1801 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1861 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1921 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1981 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2041 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2101 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2161 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2221 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2281 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2341 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2401 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2461 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2521 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2581 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2641 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2701 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2761 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2821 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     2881 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2941 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     3001 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     3061 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct
     3121 cactaaaggg aacaaaagct ggagctccgg atgcaagggt tcgaatccct tagctctcat
     3181 tattttttgc tttttctctt gaggtcacat gatcgcaaaa tggcaaatgg cacgtgaagc
     3241 tgtcgatatt ggggaactgt ggtggttggc aaatgactaa ttaagttagt caaggcgcca
     3301 tcctcatgaa aactgtgtaa cataataacc gaagtgtcga aaaggtggca ccttgtccaa
     3361 ttgaacacgc tcgatgaaaa aaataagata tatataaggt taagtaaagc gtctgttaga
     3421 aaggaagttt ttcctttttc ttgctctctt gtcttttcat ctactatttc cttcgtgtaa
     3481 tacagggtcg tcagatacat agatacaatt ctattacccc catccatact ctagaactag
     3541 tggatccatg gtggttttgg ctgcttctat cactacgcgt caggggaaac ctctcttgtc
     3601 caggcaattc aaagatctgt ccaaggatcg agtcctagag ttgctgtcca atttccagaa
     3661 cctagtctcc gagatctcgt cagaccacac atttgtcgag gacaagcacg tccgctacgt
     3721 gtataggccc tttgataact actacattat tcttatcaca aaccgccaga gcaacatctt
     3781 agaagatctg gagaccctaa ggctcttctc aagagtgatt cctgaatatt gccgagcctt
     3841 agaagagaat gaaatatctg agcactgttt tgatttgatt tttgcatttg acgaaattgt
     3901 tgccttggga taccgggaga atgttaacct ggcacagatc agaaccttca cagaaatgga
     3961 ttcccatgag gagaaggtgt tcagagcagt cagagagact caagagcgtg aagccaaggc
     4021 tgagatgcgg cgtaaagcaa aagaattaca gcaggcccga agagatgcag agagacaggg
     4081 ccatcaccat caccatcact gaccggaatt ccggccgctc gagtcatgta attagttatg
     4141 tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga
     4201 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
     4261 tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata
     4321 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccggtac
     4381 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
     4441 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     4501 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     4561 tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4621 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4681 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4741 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4801 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4861 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4921 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4981 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg
     5041 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatcgaccgg tcgaggagaa
     5101 cttctagtat atctacatac ctaatattat tgccttatta aaaatggaat cccaacaatt
     5161 acatcaaaat ccacattctc ttcaaaatca attgtcctgt acttccttgt tcatgtgtgt
     5221 tcaaaaacgt tatatttata ggataattat actctatttc tcaacaagta attggttgtt
     5281 tggccgagcg gtctaaggcg cctgattcaa gaaatatctt gaccgcagtt aactgtggga
     5341 atactcaggt atcgtaagat gcaagagttc gaatctctta gcaaccatta tttttttcct
     5401 caacataacg agaacacaca ggggcgctat cgcacagaat caaattcgat gactggaaat
     5461 tttttgttaa tttcagaggt cgcctgacgc atataccttt ttcaactgaa aaattgggag
     5521 aaaaaggaaa ggtgagagcg ccggaaccgg cttttcatat agaatagaga agcgttcatg
     5581 actaaatgct tgcatcacaa tacttgaagt tgacaatatt atttaaggac ctattgtttt
     5641 ttccaatagg tggttagcaa tcgtcttact ttctaacttt tcttaccttt tacatttcag
     5701 caatatatat atatatattt caaggatata ccattctaat gtctgcccct aagaagatcg
     5761 tcgttttgcc aggtgaccac gttggtcaag aaatcacagc cgaagccatt aaggttctta
     5821 aagctatttc tgatgttcgt tccaatgtca agttcgattt cgaaaatcat ttaattggtg
     5881 gtgctgctat cgatgctaca ggtgttccac ttccagatga ggcgctggaa gcctccaaga
     5941 aggctgatgc cgttttgtta ggtgctgtgg gtggtcctaa atggggtacc ggtagtgtta
     6001 gacctgaaca aggtttacta aaaatccgta aagaacttca attgtacgcc aacttaagac
     6061 catgtaactt tgcatccgac tctcttttag acttatctcc aatcaagcca caatttgcta
     6121 aaggtactga cttcgttgtt gtcagagaat tagtgggagg tatttacttt ggtaagagaa
     6181 aggaagacga tggtgatggt gtcgcttggg atagtgaaca atacaccgtt ccagaagtgc
     6241 aaagaatcac aagaatggcc gctttcatgg ccctacaaca tgagccacca ttgcctattt
     6301 ggtccttgga taaagctaat gttttggcct cttcaagatt atggagaaaa actgtggagg
     6361 aaaccatcaa gaacgaattc cctacattga aggttcaaca tcaattgatt gattctgccg
     6421 ccatgatcct agttaagaac ccaacccacc taaatggtat tataatcacc agcaacatgt
     6481 ttggtgatat catctccgat gaagcctccg ttatcccagg ttccttgggt ttgttgccat
     6541 ctgcgtcctt ggcctctttg ccagacaaga acaccgcatt tggtttgtac gaaccatgcc
     6601 acggttctgc tccagatttg ccaaagaata aggtcaaccc tatcgccact atcttgtctg
     6661 ctgcaatgat gttgaaattg tcattgaact tgcctgaaga aggtaaggcc attgaagatg
     6721 cagttaaaaa ggttttggat gcaggtatca gaactggtga tttaggtggt tccaacagta
     6781 ccaccgaagt cggtgatgct gtcgccgaag aagttaagaa aatccttgct taaaaagatt
     6841 ctcttttttt atgatatttg tacataaact ttataaatga aattcataat agaaacgaca
     6901 cgaaattaca aaatggaata tgttcatagg gtagacgaaa ctatatacgc aatctacata
     6961 catttatcaa gaaggagaaa aaggaggatg taaaggaata caggtaagca aattgatact
     7021 aatggctcaa cgtgataagg aaaaagaatt gcactttaac attaatattg acaaggagga
     7081 gggcaccaca caaaaagtta ggtgtaacag aaaatcatga aactatgatt cctaatttat
     7141 at
//
