LOCUS       sequence-200049-        7142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chimaera 10.
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-10
FEATURES             Location/Qualifiers
     source          1..7142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(97..116)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     216..238
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(276..294)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    336..839
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        876..980
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(981..1049,1050..1841)
                     /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(1199..1218)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2012..2600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2501..2520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2754..2771
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2888..2909
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2924..2929,2930..2947,2948..2954)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2962..2978
                     /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     2967..2989
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2986..3002
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2986..3002
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3021..3041
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3023..3041
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3092..3441
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3442..3458
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3442..3458
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3994..4011
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4036..4283
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4052..4070)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4301..4320)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4302..4320)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4330..4347)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4330..4346)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4339..4361)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4487..4942
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4574..4593)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4784..4805
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5243..5650
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5651..6745
                     /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 ttcttgaacc atttcccata atggtgaaag ttccctcaag aattttactc tgtcagaaac
       61 ggccttaacg acgtagtcga tatggtgcac tctcagtaca atctgctctg atgccgcata
      121 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
      181 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
      241 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
      301 ggttaatgtc atgataataa tggtttctta ggacggatcg cttgcctgta acttacacgc
      361 gcctcgtatc ttttaatgat ggaataattt gggaatttac tctgtgttta tttattttta
      421 tgttttgtat ttggatttta gaaagtaaat aaagaaggta gaagagttac ggaatgaaga
      481 aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag cgtactttac atatatattt
      541 attagacaag aaaagcagat taaatagata tacattcgat taacgataag taaaatgtaa
      601 aatcacagga ttttcgtgtg tggtcttcta cacagacaag atgaaacaat tcggcattaa
      661 tacctgagag caggaagagc aagataaaag gtagtatttg ttggcgatcc ccctagagtc
      721 ttttacatct tcggaaaaca aaaactattt tttctttaat ttcttttttt actttctatt
      781 tttaatttat atatttatat taaaaaattt aaattataat tatttttata gcacgtgatg
      841 aaaaggaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      901 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      961 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1021 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1081 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1141 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1201 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     1261 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1321 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1381 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1441 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1501 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     1561 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1621 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1681 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     1741 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1801 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1861 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1921 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1981 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2041 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2101 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2161 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2221 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2281 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2341 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2401 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2461 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2521 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2581 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2641 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2701 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2761 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2821 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2881 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2941 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     3001 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctcc
     3061 ggatgcaagg gttcgaatcc cttagctctc attatttttt gctttttctc ttgaggtcac
     3121 atgatcgcaa aatggcaaat ggcacgtgaa gctgtcgata ttggggaact gtggtggttg
     3181 gcaaatgact aattaagtta gtcaaggcgc catcctcatg aaaactgtgt aacataataa
     3241 ccgaagtgtc gaaaaggtgg caccttgtcc aattgaacac gctcgatgaa aaaaataaga
     3301 tatatataag gttaagtaaa gcgtctgtta gaaaggaagt ttttcctttt tcttgctctc
     3361 ttgtcttttc atctactatt tccttcgtgt aatacagggt cgtcagatac atagatacaa
     3421 ttctattacc cccatccata ctctagaact agtggatcca tggtggtttt ggctgcttct
     3481 atcactacgc gtcaggggaa acctctcttg tccaggcaat tcaaagatct gtccaaggat
     3541 cgagtcctag agttgctgtc caatttccag aacctagtct ccgagatctc gtcagaccac
     3601 acatttgtcg aggacaagca cgtccgctac gtgtataggc cctttgataa ctactacatt
     3661 attcttatca caaaccgcca gtccaatatt atcaaggatt tggccacatt aaatctgttt
     3721 tcacagacga tcaactccta cttgtctagt ttccaagacc aagagatttt tcacaatgcc
     3781 tttgagatct tgtcctcgtt cgacgaaatc gtgtcaatgg gcgggtacaa agagaacttg
     3841 tcctttactc aagtgcaaac atacttgtct atggagtcgc acgaagaaag agtgttcaga
     3901 gcagtcagag agactcaaga gcgtgaagcc aaggctgaga tgaagcgccg tgctaaggaa
     3961 attgctagaa aagaacatga aaggaaacac gggcatcacc atcaccatca ctgaccggaa
     4021 ttccggccgc tcgagtcatg taattagtta tgtcacgctt acattcacgc cctcccccca
     4081 catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt
     4141 tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     4201 tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg
     4261 gacgctcgaa ggctttaatt tgcggccggt acccaattcg ccctatagtg agtcgtatta
     4321 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     4381 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     4441 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc
     4501 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4561 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4621 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4681 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4741 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4801 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4861 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4921 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4981 tttcacaccg catatcgacc ggtcgaggag aacttctagt atatctacat acctaatatt
     5041 attgccttat taaaaatgga atcccaacaa ttacatcaaa atccacattc tcttcaaaat
     5101 caattgtcct gtacttcctt gttcatgtgt gttcaaaaac gttatattta taggataatt
     5161 atactctatt tctcaacaag taattggttg tttggccgag cggtctaagg cgcctgattc
     5221 aagaaatatc ttgaccgcag ttaactgtgg gaatactcag gtatcgtaag atgcaagagt
     5281 tcgaatctct tagcaaccat tatttttttc ctcaacataa cgagaacaca caggggcgct
     5341 atcgcacaga atcaaattcg atgactggaa attttttgtt aatttcagag gtcgcctgac
     5401 gcatatacct ttttcaactg aaaaattggg agaaaaagga aaggtgagag cgccggaacc
     5461 ggcttttcat atagaataga gaagcgttca tgactaaatg cttgcatcac aatacttgaa
     5521 gttgacaata ttatttaagg acctattgtt ttttccaata ggtggttagc aatcgtctta
     5581 ctttctaact tttcttacct tttacatttc agcaatatat atatatatat ttcaaggata
     5641 taccattcta atgtctgccc ctaagaagat cgtcgttttg ccaggtgacc acgttggtca
     5701 agaaatcaca gccgaagcca ttaaggttct taaagctatt tctgatgttc gttccaatgt
     5761 caagttcgat ttcgaaaatc atttaattgg tggtgctgct atcgatgcta caggtgttcc
     5821 acttccagat gaggcgctgg aagcctccaa gaaggctgat gccgttttgt taggtgctgt
     5881 gggtggtcct aaatggggta ccggtagtgt tagacctgaa caaggtttac taaaaatccg
     5941 taaagaactt caattgtacg ccaacttaag accatgtaac tttgcatccg actctctttt
     6001 agacttatct ccaatcaagc cacaatttgc taaaggtact gacttcgttg ttgtcagaga
     6061 attagtggga ggtatttact ttggtaagag aaaggaagac gatggtgatg gtgtcgcttg
     6121 ggatagtgaa caatacaccg ttccagaagt gcaaagaatc acaagaatgg ccgctttcat
     6181 ggccctacaa catgagccac cattgcctat ttggtccttg gataaagcta atgttttggc
     6241 ctcttcaaga ttatggagaa aaactgtgga ggaaaccatc aagaacgaat tccctacatt
     6301 gaaggttcaa catcaattga ttgattctgc cgccatgatc ctagttaaga acccaaccca
     6361 cctaaatggt attataatca ccagcaacat gtttggtgat atcatctccg atgaagcctc
     6421 cgttatccca ggttccttgg gtttgttgcc atctgcgtcc ttggcctctt tgccagacaa
     6481 gaacaccgca tttggtttgt acgaaccatg ccacggttct gctccagatt tgccaaagaa
     6541 taaggtcaac cctatcgcca ctatcttgtc tgctgcaatg atgttgaaat tgtcattgaa
     6601 cttgcctgaa gaaggtaagg ccattgaaga tgcagttaaa aaggttttgg atgcaggtat
     6661 cagaactggt gatttaggtg gttccaacag taccaccgaa gtcggtgatg ctgtcgccga
     6721 agaagttaag aaaatccttg cttaaaaaga ttctcttttt ttatgatatt tgtacataaa
     6781 ctttataaat gaaattcata atagaaacga cacgaaatta caaaatggaa tatgttcata
     6841 gggtagacga aactatatac gcaatctaca tacatttatc aagaaggaga aaaaggagga
     6901 tgtaaaggaa tacaggtaag caaattgata ctaatggctc aacgtgataa ggaaaaagaa
     6961 ttgcacttta acattaatat tgacaaggag gagggcacca cacaaaaagt taggtgtaac
     7021 agaaaatcat gaaactatga ttcctaattt atatattgga ggattttctc taaaaaaaaa
     7081 aaaatacaac aaataaaaaa cactcaatga cctgaccatt tgatggagtt taagtcaata
     7141 cc
//
