LOCUS       sequence-200047-        7142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chimaera 9.
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-9
FEATURES             Location/Qualifiers
     source          1..7142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(213..232)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     332..354
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(392..410)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    452..955
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        992..1096
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1097..1165,1166..1957)
                     /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(1315..1334)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2128..2716
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2617..2636
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2870..2887
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3004..3025
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3040..3045,3046..3063,3064..3070)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3078..3094
                     /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     3083..3105
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3102..3118
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3102..3118
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3137..3157
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3139..3157
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3208..3557
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3558..3574
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3558..3574
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4110..4127
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4152..4399
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4168..4186)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4417..4436)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4418..4436)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4446..4463)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4446..4462)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4455..4477)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4603..5058
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4690..4709)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4900..4921
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5359..5766
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5767..6861
                     /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 tcatgaaact atgattccta atttatatat tggaggattt tctctaaaaa aaaaaaaata
       61 caacaaataa aaaacactca atgacctgac catttgatgg agtttaagtc aataccttct
      121 tgaaccattt cccataatgg tgaaagttcc ctcaagaatt ttactctgtc agaaacggcc
      181 ttaacgacgt agtcgatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
      241 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
      301 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
      361 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
      421 aatgtcatga taataatggt ttcttaggac ggatcgcttg cctgtaactt acacgcgcct
      481 cgtatctttt aatgatggaa taatttggga atttactctg tgtttattta tttttatgtt
      541 ttgtatttgg attttagaaa gtaaataaag aaggtagaag agttacggaa tgaagaaaaa
      601 aaaataaaca aaggtttaaa aaatttcaac aaaaagcgta ctttacatat atatttatta
      661 gacaagaaaa gcagattaaa tagatataca ttcgattaac gataagtaaa atgtaaaatc
      721 acaggatttt cgtgtgtggt cttctacaca gacaagatga aacaattcgg cattaatacc
      781 tgagagcagg aagagcaaga taaaaggtag tatttgttgg cgatccccct agagtctttt
      841 acatcttcgg aaaacaaaaa ctattttttc tttaatttct ttttttactt tctattttta
      901 atttatatat ttatattaaa aaatttaaat tataattatt tttatagcac gtgatgaaaa
      961 ggacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1021 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1081 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1141 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1201 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1261 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1321 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1381 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1441 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     1501 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     1561 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1621 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     1681 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1741 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1801 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     1861 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1921 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1981 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2041 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2101 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2161 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2221 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2281 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2341 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2401 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2461 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2521 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2581 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2641 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2701 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2761 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2821 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2881 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     2941 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     3001 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     3061 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     3121 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctccggat
     3181 gcaagggttc gaatccctta gctctcatta ttttttgctt tttctcttga ggtcacatga
     3241 tcgcaaaatg gcaaatggca cgtgaagctg tcgatattgg ggaactgtgg tggttggcaa
     3301 atgactaatt aagttagtca aggcgccatc ctcatgaaaa ctgtgtaaca taataaccga
     3361 agtgtcgaaa aggtggcacc ttgtccaatt gaacacgctc gatgaaaaaa ataagatata
     3421 tataaggtta agtaaagcgt ctgttagaaa ggaagttttt cctttttctt gctctcttgt
     3481 cttttcatct actatttcct tcgtgtaata cagggtcgtc agatacatag atacaattct
     3541 attaccccca tccatactct agaactagtg gatccatggt ggttttggct gcttctatca
     3601 ctacgcgtca ggggaaacct ctcttgtcca ggcaattcaa agatctgtcc aaggatcgag
     3661 tcctagagtt gctgtccaat ttccagaacc tagtctccga gatctcgtca gaccacacat
     3721 ttgtcgagga caagcacgtc cgctacgtgt ataggccctt tgataactac tacattattc
     3781 ttatcacaaa ccgccagtcc aatattatca aggatttggc cacattaaat ctgttttcac
     3841 agacgatcaa ctcctacttg tctagtttcc aagaccaaga gatttttcac aatgcctttg
     3901 agatcttgtc ctcgttcgac gaaatcgtgt caatgggcgg gtacaaagag aacgttaacc
     3961 tggcacagat cagaaccttc acagaaatgg agtcgcacga agaaagaatc caggaaatta
     4021 ttgagcgtaa taaagagatc gaagctactg aggagaggaa gcgccgtgct aaggaaattg
     4081 ctagaaaaga acatgaaagg aaacacgggc atcaccatca ccatcactga ccggaattcc
     4141 ggccgctcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     4201 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     4261 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     4321 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     4381 ctcgaaggct ttaatttgcg gccggtaccc aattcgccct atagtgagtc gtattacgcg
     4441 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     4501 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     4561 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     4621 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4681 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4741 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4801 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4861 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4921 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4981 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5041 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     5101 acaccgcata tcgaccggtc gaggagaact tctagtatat ctacatacct aatattattg
     5161 ccttattaaa aatggaatcc caacaattac atcaaaatcc acattctctt caaaatcaat
     5221 tgtcctgtac ttccttgttc atgtgtgttc aaaaacgtta tatttatagg ataattatac
     5281 tctatttctc aacaagtaat tggttgtttg gccgagcggt ctaaggcgcc tgattcaaga
     5341 aatatcttga ccgcagttaa ctgtgggaat actcaggtat cgtaagatgc aagagttcga
     5401 atctcttagc aaccattatt tttttcctca acataacgag aacacacagg ggcgctatcg
     5461 cacagaatca aattcgatga ctggaaattt tttgttaatt tcagaggtcg cctgacgcat
     5521 ataccttttt caactgaaaa attgggagaa aaaggaaagg tgagagcgcc ggaaccggct
     5581 tttcatatag aatagagaag cgttcatgac taaatgcttg catcacaata cttgaagttg
     5641 acaatattat ttaaggacct attgtttttt ccaataggtg gttagcaatc gtcttacttt
     5701 ctaacttttc ttacctttta catttcagca atatatatat atatatttca aggatatacc
     5761 attctaatgt ctgcccctaa gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa
     5821 atcacagccg aagccattaa ggttcttaaa gctatttctg atgttcgttc caatgtcaag
     5881 ttcgatttcg aaaatcattt aattggtggt gctgctatcg atgctacagg tgttccactt
     5941 ccagatgagg cgctggaagc ctccaagaag gctgatgccg ttttgttagg tgctgtgggt
     6001 ggtcctaaat ggggtaccgg tagtgttaga cctgaacaag gtttactaaa aatccgtaaa
     6061 gaacttcaat tgtacgccaa cttaagacca tgtaactttg catccgactc tcttttagac
     6121 ttatctccaa tcaagccaca atttgctaaa ggtactgact tcgttgttgt cagagaatta
     6181 gtgggaggta tttactttgg taagagaaag gaagacgatg gtgatggtgt cgcttgggat
     6241 agtgaacaat acaccgttcc agaagtgcaa agaatcacaa gaatggccgc tttcatggcc
     6301 ctacaacatg agccaccatt gcctatttgg tccttggata aagctaatgt tttggcctct
     6361 tcaagattat ggagaaaaac tgtggaggaa accatcaaga acgaattccc tacattgaag
     6421 gttcaacatc aattgattga ttctgccgcc atgatcctag ttaagaaccc aacccaccta
     6481 aatggtatta taatcaccag caacatgttt ggtgatatca tctccgatga agcctccgtt
     6541 atcccaggtt ccttgggttt gttgccatct gcgtccttgg cctctttgcc agacaagaac
     6601 accgcatttg gtttgtacga accatgccac ggttctgctc cagatttgcc aaagaataag
     6661 gtcaacccta tcgccactat cttgtctgct gcaatgatgt tgaaattgtc attgaacttg
     6721 cctgaagaag gtaaggccat tgaagatgca gttaaaaagg ttttggatgc aggtatcaga
     6781 actggtgatt taggtggttc caacagtacc accgaagtcg gtgatgctgt cgccgaagaa
     6841 gttaagaaaa tccttgctta aaaagattct ctttttttat gatatttgta cataaacttt
     6901 ataaatgaaa ttcataatag aaacgacacg aaattacaaa atggaatatg ttcatagggt
     6961 agacgaaact atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatgta
     7021 aaggaataca ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa aaagaattgc
     7081 actttaacat taatattgac aaggaggagg gcaccacaca aaaagttagg tgtaacagaa
     7141 aa
//
