LOCUS       sequence-200053-        7142 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Chimaera 14.
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 Apr 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-14
FEATURES             Location/Qualifiers
     source          1..7142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(87..106)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     206..228
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(266..284)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    326..829
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        866..970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(971..1039,1040..1831)
                     /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(1189..1208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2002..2590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2491..2510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2744..2761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2878..2899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2914..2919,2920..2937,2938..2944)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2952..2968
                     /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     2957..2979
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2976..2992
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2976..2992
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3011..3031
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3013..3031
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3082..3431
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3432..3448
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3432..3448
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3984..4001
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4026..4273
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4042..4060)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4291..4310)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4292..4310)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4320..4337)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4320..4336)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4329..4351)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4477..4932
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4564..4583)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4774..4795
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5233..5640
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5641..6735
                     /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 atttcccata atggtgaaag ttccctcaag aattttactc tgtcagaaac ggccttaacg
       61 acgtagtcga tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
      121 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
      181 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
      241 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
      301 atgataataa tggtttctta ggacggatcg cttgcctgta acttacacgc gcctcgtatc
      361 ttttaatgat ggaataattt gggaatttac tctgtgttta tttattttta tgttttgtat
      421 ttggatttta gaaagtaaat aaagaaggta gaagagttac ggaatgaaga aaaaaaaata
      481 aacaaaggtt taaaaaattt caacaaaaag cgtactttac atatatattt attagacaag
      541 aaaagcagat taaatagata tacattcgat taacgataag taaaatgtaa aatcacagga
      601 ttttcgtgtg tggtcttcta cacagacaag atgaaacaat tcggcattaa tacctgagag
      661 caggaagagc aagataaaag gtagtatttg ttggcgatcc ccctagagtc ttttacatct
      721 tcggaaaaca aaaactattt tttctttaat ttcttttttt actttctatt tttaatttat
      781 atatttatat taaaaaattt aaattataat tatttttata gcacgtgatg aaaaggaccc
      841 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      901 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      961 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1021 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1081 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1141 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1201 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1261 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1321 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1381 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1441 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1501 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1561 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1621 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     1681 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     1741 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1801 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1861 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1921 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1981 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2041 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2101 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2161 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2221 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2281 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2341 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2401 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2461 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2521 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2581 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2641 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2701 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     2761 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2821 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2881 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2941 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     3001 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctcc ggatgcaagg
     3061 gttcgaatcc cttagctctc attatttttt gctttttctc ttgaggtcac atgatcgcaa
     3121 aatggcaaat ggcacgtgaa gctgtcgata ttggggaact gtggtggttg gcaaatgact
     3181 aattaagtta gtcaaggcgc catcctcatg aaaactgtgt aacataataa ccgaagtgtc
     3241 gaaaaggtgg caccttgtcc aattgaacac gctcgatgaa aaaaataaga tatatataag
     3301 gttaagtaaa gcgtctgtta gaaaggaagt ttttcctttt tcttgctctc ttgtcttttc
     3361 atctactatt tccttcgtgt aatacagggt cgtcagatac atagatacaa ttctattacc
     3421 cccatccata ctctagaact agtggatcca tggtggtttt ggctgcttct atcactacgc
     3481 gtcaggggaa acctctcttg tccaggcaat tcaaagatct gtccaaggat cgagtcctag
     3541 agttgctgtc caatttccag aacctagtct ccgagatctc gtcagaccac acatttgtcg
     3601 aggacaagca cgtccgctac gtgtataggc cctttgataa ctactacatt attcttatca
     3661 caaaccgcca gagcaacatc ttagaagatc tggagaccct aaggctcttc tcaagagtga
     3721 ttcctgaata ttgccgagcc ttagaagaga atgaaatatc tgagcactgt tttgatttga
     3781 tttttgcatt tgacgaaatt gttgccttgg gagggtacaa agagaacttg tcctttactc
     3841 aagtgcaaac atacttgtct atggagtcgc acgaagaaag aatccaggaa attattgagc
     3901 gtaataaaga gatcgaagct actgaggaga ggaagcgccg tgctaaggaa attgctagaa
     3961 aagaacatga aaggaaacac gggcatcacc atcaccatca ctgaccggaa ttccggccgc
     4021 tcgagtcatg taattagtta tgtcacgctt acattcacgc cctcccccca catccgctct
     4081 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt
     4141 tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg
     4201 cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgaa
     4261 ggctttaatt tgcggccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     4321 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4381 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4441 ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa
     4501 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4561 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4621 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4681 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4741 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4801 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4861 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4921 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4981 catatcgacc ggtcgaggag aacttctagt atatctacat acctaatatt attgccttat
     5041 taaaaatgga atcccaacaa ttacatcaaa atccacattc tcttcaaaat caattgtcct
     5101 gtacttcctt gttcatgtgt gttcaaaaac gttatattta taggataatt atactctatt
     5161 tctcaacaag taattggttg tttggccgag cggtctaagg cgcctgattc aagaaatatc
     5221 ttgaccgcag ttaactgtgg gaatactcag gtatcgtaag atgcaagagt tcgaatctct
     5281 tagcaaccat tatttttttc ctcaacataa cgagaacaca caggggcgct atcgcacaga
     5341 atcaaattcg atgactggaa attttttgtt aatttcagag gtcgcctgac gcatatacct
     5401 ttttcaactg aaaaattggg agaaaaagga aaggtgagag cgccggaacc ggcttttcat
     5461 atagaataga gaagcgttca tgactaaatg cttgcatcac aatacttgaa gttgacaata
     5521 ttatttaagg acctattgtt ttttccaata ggtggttagc aatcgtctta ctttctaact
     5581 tttcttacct tttacatttc agcaatatat atatatatat ttcaaggata taccattcta
     5641 atgtctgccc ctaagaagat cgtcgttttg ccaggtgacc acgttggtca agaaatcaca
     5701 gccgaagcca ttaaggttct taaagctatt tctgatgttc gttccaatgt caagttcgat
     5761 ttcgaaaatc atttaattgg tggtgctgct atcgatgcta caggtgttcc acttccagat
     5821 gaggcgctgg aagcctccaa gaaggctgat gccgttttgt taggtgctgt gggtggtcct
     5881 aaatggggta ccggtagtgt tagacctgaa caaggtttac taaaaatccg taaagaactt
     5941 caattgtacg ccaacttaag accatgtaac tttgcatccg actctctttt agacttatct
     6001 ccaatcaagc cacaatttgc taaaggtact gacttcgttg ttgtcagaga attagtggga
     6061 ggtatttact ttggtaagag aaaggaagac gatggtgatg gtgtcgcttg ggatagtgaa
     6121 caatacaccg ttccagaagt gcaaagaatc acaagaatgg ccgctttcat ggccctacaa
     6181 catgagccac cattgcctat ttggtccttg gataaagcta atgttttggc ctcttcaaga
     6241 ttatggagaa aaactgtgga ggaaaccatc aagaacgaat tccctacatt gaaggttcaa
     6301 catcaattga ttgattctgc cgccatgatc ctagttaaga acccaaccca cctaaatggt
     6361 attataatca ccagcaacat gtttggtgat atcatctccg atgaagcctc cgttatccca
     6421 ggttccttgg gtttgttgcc atctgcgtcc ttggcctctt tgccagacaa gaacaccgca
     6481 tttggtttgt acgaaccatg ccacggttct gctccagatt tgccaaagaa taaggtcaac
     6541 cctatcgcca ctatcttgtc tgctgcaatg atgttgaaat tgtcattgaa cttgcctgaa
     6601 gaaggtaagg ccattgaaga tgcagttaaa aaggttttgg atgcaggtat cagaactggt
     6661 gatttaggtg gttccaacag taccaccgaa gtcggtgatg ctgtcgccga agaagttaag
     6721 aaaatccttg cttaaaaaga ttctcttttt ttatgatatt tgtacataaa ctttataaat
     6781 gaaattcata atagaaacga cacgaaatta caaaatggaa tatgttcata gggtagacga
     6841 aactatatac gcaatctaca tacatttatc aagaaggaga aaaaggagga tgtaaaggaa
     6901 tacaggtaag caaattgata ctaatggctc aacgtgataa ggaaaaagaa ttgcacttta
     6961 acattaatat tgacaaggag gagggcacca cacaaaaagt taggtgtaac agaaaatcat
     7021 gaaactatga ttcctaattt atatattgga ggattttctc taaaaaaaaa aaaatacaac
     7081 aaataaaaaa cactcaatga cctgaccatt tgatggagtt taagtcaata ccttcttgaa
     7141 cc
//
