LOCUS       sequence-200052-        7145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chimaera 13.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7145)
  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 7145)
  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-13
FEATURES             Location/Qualifiers
     source          1..7145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(189..208)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     308..330
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(368..386)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    428..931
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        968..1072
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1073..1141,1142..1933)
                     /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(1291..1310)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2104..2692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2593..2612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2846..2863
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2980..3001
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3016..3021,3022..3039,3040..3046)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3054..3070
                     /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     3059..3081
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3078..3094
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3078..3094
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3113..3133
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3115..3133
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3184..3533
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3534..3550
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3534..3550
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4089..4106
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4131..4378
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4147..4165)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4396..4415)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4397..4415)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4425..4442)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4425..4441)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4434..4456)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4582..5037
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4669..4688)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4879..4900
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5338..5745
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5746..6840
                     /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 atatattgga ggattttctc taaaaaaaaa aaaatacaac aaataaaaaa cactcaatga
       61 cctgaccatt tgatggagtt taagtcaata ccttcttgaa ccatttccca taatggtgaa
      121 agttccctca agaattttac tctgtcagaa acggccttaa cgacgtagtc gatatggtgc
      181 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
      241 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
      301 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
      361 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
      421 taggacggat cgcttgcctg taacttacac gcgcctcgta tcttttaatg atggaataat
      481 ttgggaattt actctgtgtt tatttatttt tatgttttgt atttggattt tagaaagtaa
      541 ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg tttaaaaaat
      601 ttcaacaaaa agcgtacttt acatatatat ttattagaca agaaaagcag attaaataga
      661 tatacattcg attaacgata agtaaaatgt aaaatcacag gattttcgtg tgtggtcttc
      721 tacacagaca agatgaaaca attcggcatt aatacctgag agcaggaaga gcaagataaa
      781 aggtagtatt tgttggcgat ccccctagag tcttttacat cttcggaaaa caaaaactat
      841 tttttcttta atttcttttt ttactttcta tttttaattt atatatttat attaaaaaat
      901 ttaaattata attattttta tagcacgtga tgaaaaggac ccaggtggca cttttcgggg
      961 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1021 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1081 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1141 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1201 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1261 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1321 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1381 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1441 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1501 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1561 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1621 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1681 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1741 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1801 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1861 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     1921 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     1981 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2041 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2101 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2161 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2221 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2281 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2341 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2401 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2461 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2521 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2581 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2641 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2701 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2761 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2821 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     2881 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     2941 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     3001 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     3061 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     3121 ccctcactaa agggaacaaa agctggagct ccggatgcaa gggttcgaat cccttagctc
     3181 tcattatttt ttgctttttc tcttgaggtc acatgatcgc aaaatggcaa atggcacgtg
     3241 aagctgtcga tattggggaa ctgtggtggt tggcaaatga ctaattaagt tagtcaaggc
     3301 gccatcctca tgaaaactgt gtaacataat aaccgaagtg tcgaaaaggt ggcaccttgt
     3361 ccaattgaac acgctcgatg aaaaaaataa gatatatata aggttaagta aagcgtctgt
     3421 tagaaaggaa gtttttcctt tttcttgctc tcttgtcttt tcatctacta tttccttcgt
     3481 gtaatacagg gtcgtcagat acatagatac aattctatta cccccatcca tactctagaa
     3541 ctagtggatc catggtggtt ttggctgctt ctatcactac gcgtcagggg aaacctctct
     3601 tgtccaggca attcaaagat ctgtccaagg atcgagtcct agagttgctg tccaatttcc
     3661 agaacctagt ctccgagatc tcgtcagacc acacatttgt cgaggacaag cacgtccgct
     3721 acgtgtatag gccctttgat aactactaca ttattcttat cacaaaccgc cagtccaata
     3781 ttatcaagga tttggccaca ttaaatctgt tttcacagac gatcaactcc tacttgtcta
     3841 gtttccaaga ccaagagatt tttcacaatg cctttgagat cttgtcctcg ttcgacgaaa
     3901 tcgtgtcaat gggcgggtac aaagagaacg ttaacctggc acagatcaga accttcacag
     3961 aaatggattc ccatgaggag aagatccagg aaattattga gcgtaataaa gagatcgaag
     4021 ctactgagga gaggcggcgt aaagcaaaag aattacagca ggcccgaaga gatgcagaga
     4081 gacagggcca tcaccatcac catcactgac cggaattccg gccgctcgag tcatgtaatt
     4141 agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg
     4201 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
     4261 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
     4321 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg
     4381 ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     4441 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     4501 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     4561 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4621 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4681 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4741 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4801 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4861 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4921 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4981 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
     5041 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat cgaccggtcg
     5101 aggagaactt ctagtatatc tacataccta atattattgc cttattaaaa atggaatccc
     5161 aacaattaca tcaaaatcca cattctcttc aaaatcaatt gtcctgtact tccttgttca
     5221 tgtgtgttca aaaacgttat atttatagga taattatact ctatttctca acaagtaatt
     5281 ggttgtttgg ccgagcggtc taaggcgcct gattcaagaa atatcttgac cgcagttaac
     5341 tgtgggaata ctcaggtatc gtaagatgca agagttcgaa tctcttagca accattattt
     5401 ttttcctcaa cataacgaga acacacaggg gcgctatcgc acagaatcaa attcgatgac
     5461 tggaaatttt ttgttaattt cagaggtcgc ctgacgcata tacctttttc aactgaaaaa
     5521 ttgggagaaa aaggaaaggt gagagcgccg gaaccggctt ttcatataga atagagaagc
     5581 gttcatgact aaatgcttgc atcacaatac ttgaagttga caatattatt taaggaccta
     5641 ttgttttttc caataggtgg ttagcaatcg tcttactttc taacttttct taccttttac
     5701 atttcagcaa tatatatata tatatttcaa ggatatacca ttctaatgtc tgcccctaag
     5761 aagatcgtcg ttttgccagg tgaccacgtt ggtcaagaaa tcacagccga agccattaag
     5821 gttcttaaag ctatttctga tgttcgttcc aatgtcaagt tcgatttcga aaatcattta
     5881 attggtggtg ctgctatcga tgctacaggt gttccacttc cagatgaggc gctggaagcc
     5941 tccaagaagg ctgatgccgt tttgttaggt gctgtgggtg gtcctaaatg gggtaccggt
     6001 agtgttagac ctgaacaagg tttactaaaa atccgtaaag aacttcaatt gtacgccaac
     6061 ttaagaccat gtaactttgc atccgactct cttttagact tatctccaat caagccacaa
     6121 tttgctaaag gtactgactt cgttgttgtc agagaattag tgggaggtat ttactttggt
     6181 aagagaaagg aagacgatgg tgatggtgtc gcttgggata gtgaacaata caccgttcca
     6241 gaagtgcaaa gaatcacaag aatggccgct ttcatggccc tacaacatga gccaccattg
     6301 cctatttggt ccttggataa agctaatgtt ttggcctctt caagattatg gagaaaaact
     6361 gtggaggaaa ccatcaagaa cgaattccct acattgaagg ttcaacatca attgattgat
     6421 tctgccgcca tgatcctagt taagaaccca acccacctaa atggtattat aatcaccagc
     6481 aacatgtttg gtgatatcat ctccgatgaa gcctccgtta tcccaggttc cttgggtttg
     6541 ttgccatctg cgtccttggc ctctttgcca gacaagaaca ccgcatttgg tttgtacgaa
     6601 ccatgccacg gttctgctcc agatttgcca aagaataagg tcaaccctat cgccactatc
     6661 ttgtctgctg caatgatgtt gaaattgtca ttgaacttgc ctgaagaagg taaggccatt
     6721 gaagatgcag ttaaaaaggt tttggatgca ggtatcagaa ctggtgattt aggtggttcc
     6781 aacagtacca ccgaagtcgg tgatgctgtc gccgaagaag ttaagaaaat ccttgcttaa
     6841 aaagattctc tttttttatg atatttgtac ataaacttta taaatgaaat tcataataga
     6901 aacgacacga aattacaaaa tggaatatgt tcatagggta gacgaaacta tatacgcaat
     6961 ctacatacat ttatcaagaa ggagaaaaag gaggatgtaa aggaatacag gtaagcaaat
     7021 tgatactaat ggctcaacgt gataaggaaa aagaattgca ctttaacatt aatattgaca
     7081 aggaggaggg caccacacaa aaagttaggt gtaacagaaa atcatgaaac tatgattcct
     7141 aattt
//
