LOCUS       sequence-200166-        7166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Swapped helices.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7166)
  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 7166)
  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 Ret2LD+delta-C2-alpha
FEATURES             Location/Qualifiers
     source          1..7166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(91..110)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     210..232
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(270..288)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    330..833
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        870..974
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(975..1043,1044..1835)
                     /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(1193..1212)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2006..2594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2495..2514
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2748..2765
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2882..2903
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2918..2923,2924..2941,2942..2948)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2956..2972
                     /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     2961..2983
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2980..2996
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2980..2996
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3015..3035
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3017..3035
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3086..3435
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3436..3452
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3436..3452
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3991..4008
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4035..4051)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4036..4052)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4054..4301
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4070..4088)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4319..4338)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4320..4338)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4348..4365)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4348..4364)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4357..4379)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4505..4960
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4592..4611)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4802..4823
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5261..5668
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5669..6763
                     /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 aaccatttcc cataatggtg aaagttccct caagaatttt actctgtcag aaacggcctt
       61 aacgacgtag tcgatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
      121 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
      181 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
      241 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
      301 tgtcatgata ataatggttt cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg
      361 tatcttttaa tgatggaata atttgggaat ttactctgtg tttatttatt tttatgtttt
      421 gtatttggat tttagaaagt aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa
      481 aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact ttacatatat atttattaga
      541 caagaaaagc agattaaata gatatacatt cgattaacga taagtaaaat gtaaaatcac
      601 aggattttcg tgtgtggtct tctacacaga caagatgaaa caattcggca ttaatacctg
      661 agagcaggaa gagcaagata aaaggtagta tttgttggcg atccccctag agtcttttac
      721 atcttcggaa aacaaaaact attttttctt taatttcttt ttttactttc tatttttaat
      781 ttatatattt atattaaaaa atttaaatta taattatttt tatagcacgt gatgaaaagg
      841 acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
      901 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      961 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1021 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1081 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1141 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1201 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1261 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1321 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1381 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1441 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1501 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     1561 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1621 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1681 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1741 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1801 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1861 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1921 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1981 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2041 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2101 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2161 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2221 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2281 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2341 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2401 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2461 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2521 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2581 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2641 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2701 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2761 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2821 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2881 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     2941 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     3001 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctccggatgc
     3061 aagggttcga atcccttagc tctcattatt ttttgctttt tctcttgagg tcacatgatc
     3121 gcaaaatggc aaatggcacg tgaagctgtc gatattgggg aactgtggtg gttggcaaat
     3181 gactaattaa gttagtcaag gcgccatcct catgaaaact gtgtaacata ataaccgaag
     3241 tgtcgaaaag gtggcacctt gtccaattga acacgctcga tgaaaaaaat aagatatata
     3301 taaggttaag taaagcgtct gttagaaagg aagtttttcc tttttcttgc tctcttgtct
     3361 tttcatctac tatttccttc gtgtaataca gggtcgtcag atacatagat acaattctat
     3421 tacccccatc catactctag aactagtgga tccatggtgg ttttggctgc ttctatcact
     3481 acgcgtcagg ggaaacctct cttgtccagg caattcaaag atctgtccaa ggatcgagtc
     3541 ctagagttgc tgtccaattt ccagaaccta gtctccgaga tctcgtcaga ccacacattt
     3601 gtcgaggaca agcacgtccg ctacgtgtat aggccctttg ataactacta cattattctt
     3661 atcacaaacc gccagtccaa tattatcaag gatttggcca cattaaatct gttttcacag
     3721 acgatcaact cctacttgtc tagtttccaa gaccaagaga tttttcacaa tgcctttgag
     3781 atcttgtcct cgttcgacga aatcgtgtca atgggcgggt accgggagaa tgttaacctg
     3841 gcacagatca gaaccttcac agaaatggat tcccatgagg agaaggtgtt cagagcagtc
     3901 agagagactc aagagcgtga agccaaggct gagatgcggc gtaaagcaaa agaattacag
     3961 caggcccgaa gagatgcaga gagacagggc catcaccatc accatcactg accggaattc
     4021 gatatcaagc ttatcgatac cgtcgacctc gagtcatgta attagttatg tcacgcttac
     4081 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     4141 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     4201 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     4261 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccggtac ccaattcgcc
     4321 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     4381 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     4441 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     4501 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4561 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4621 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4681 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4741 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4801 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4861 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4921 tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     4981 ttacgcatct gtgcggtatt tcacaccgca tatcgaccgg tcgaggagaa cttctagtat
     5041 atctacatac ctaatattat tgccttatta aaaatggaat cccaacaatt acatcaaaat
     5101 ccacattctc ttcaaaatca attgtcctgt acttccttgt tcatgtgtgt tcaaaaacgt
     5161 tatatttata ggataattat actctatttc tcaacaagta attggttgtt tggccgagcg
     5221 gtctaaggcg cctgattcaa gaaatatctt gaccgcagtt aactgtggga atactcaggt
     5281 atcgtaagat gcaagagttc gaatctctta gcaaccatta tttttttcct caacataacg
     5341 agaacacaca ggggcgctat cgcacagaat caaattcgat gactggaaat tttttgttaa
     5401 tttcagaggt cgcctgacgc atataccttt ttcaactgaa aaattgggag aaaaaggaaa
     5461 ggtgagagcg ccggaaccgg cttttcatat agaatagaga agcgttcatg actaaatgct
     5521 tgcatcacaa tacttgaagt tgacaatatt atttaaggac ctattgtttt ttccaatagg
     5581 tggttagcaa tcgtcttact ttctaacttt tcttaccttt tacatttcag caatatatat
     5641 atatatattt caaggatata ccattctaat gtctgcccct aagaagatcg tcgttttgcc
     5701 aggtgaccac gttggtcaag aaatcacagc cgaagccatt aaggttctta aagctatttc
     5761 tgatgttcgt tccaatgtca agttcgattt cgaaaatcat ttaattggtg gtgctgctat
     5821 cgatgctaca ggtgttccac ttccagatga ggcgctggaa gcctccaaga aggctgatgc
     5881 cgttttgtta ggtgctgtgg gtggtcctaa atggggtacc ggtagtgtta gacctgaaca
     5941 aggtttacta aaaatccgta aagaacttca attgtacgcc aacttaagac catgtaactt
     6001 tgcatccgac tctcttttag acttatctcc aatcaagcca caatttgcta aaggtactga
     6061 cttcgttgtt gtcagagaat tagtgggagg tatttacttt ggtaagagaa aggaagacga
     6121 tggtgatggt gtcgcttggg atagtgaaca atacaccgtt ccagaagtgc aaagaatcac
     6181 aagaatggcc gctttcatgg ccctacaaca tgagccacca ttgcctattt ggtccttgga
     6241 taaagctaat gttttggcct cttcaagatt atggagaaaa actgtggagg aaaccatcaa
     6301 gaacgaattc cctacattga aggttcaaca tcaattgatt gattctgccg ccatgatcct
     6361 agttaagaac ccaacccacc taaatggtat tataatcacc agcaacatgt ttggtgatat
     6421 catctccgat gaagcctccg ttatcccagg ttccttgggt ttgttgccat ctgcgtcctt
     6481 ggcctctttg ccagacaaga acaccgcatt tggtttgtac gaaccatgcc acggttctgc
     6541 tccagatttg ccaaagaata aggtcaaccc tatcgccact atcttgtctg ctgcaatgat
     6601 gttgaaattg tcattgaact tgcctgaaga aggtaaggcc attgaagatg cagttaaaaa
     6661 ggttttggat gcaggtatca gaactggtga tttaggtggt tccaacagta ccaccgaagt
     6721 cggtgatgct gtcgccgaag aagttaagaa aatccttgct taaaaagatt ctcttttttt
     6781 atgatatttg tacataaact ttataaatga aattcataat agaaacgaca cgaaattaca
     6841 aaatggaata tgttcatagg gtagacgaaa ctatatacgc aatctacata catttatcaa
     6901 gaaggagaaa aaggaggatg taaaggaata caggtaagca aattgatact aatggctcaa
     6961 cgtgataagg aaaaagaatt gcactttaac attaatattg acaaggagga gggcaccaca
     7021 caaaaagtta ggtgtaacag aaaatcatga aactatgatt cctaatttat atattggagg
     7081 attttctcta aaaaaaaaaa aatacaacaa ataaaaaaca ctcaatgacc tgaccatttg
     7141 atggagttta agtcaatacc ttcttg
//
