LOCUS       sequence-200165-        6992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  delta-CLD - His tagged (2-alpha helices+uHD deleted).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6992)
  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 6992)
  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 delta-CLD
FEATURES             Location/Qualifiers
     source          1..6992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(82..101)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     201..223
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(261..279)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    321..824
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        861..965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(966..1034,1035..1826)
                     /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(1184..1203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1997..2585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2486..2505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2739..2756
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2873..2894
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2909..2914,2915..2932,2933..2939)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2947..2963
                     /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     2952..2974
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2971..2987
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2971..2987
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3006..3026
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3008..3026
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3077..3426
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3427..3443
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3427..3443
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3808..3825
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3852..3868)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3853..3869)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      3871..4118
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3887..3905)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4136..4155)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4137..4155)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4165..4182)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4165..4181)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4174..4196)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4322..4777
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4409..4428)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4619..4640
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5078..5485
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5486..6580
                     /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 ccataatggt gaaagttccc tcaagaattt tactctgtca gaaacggcct taacgacgta
       61 gtcgatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
      121 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
      181 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
      241 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
      301 aataatggtt tcttaggacg gatcgcttgc ctgtaactta cacgcgcctc gtatctttta
      361 atgatggaat aatttgggaa tttactctgt gtttatttat ttttatgttt tgtatttgga
      421 ttttagaaag taaataaaga aggtagaaga gttacggaat gaagaaaaaa aaataaacaa
      481 aggtttaaaa aatttcaaca aaaagcgtac tttacatata tatttattag acaagaaaag
      541 cagattaaat agatatacat tcgattaacg ataagtaaaa tgtaaaatca caggattttc
      601 gtgtgtggtc ttctacacag acaagatgaa acaattcggc attaatacct gagagcagga
      661 agagcaagat aaaaggtagt atttgttggc gatcccccta gagtctttta catcttcgga
      721 aaacaaaaac tattttttct ttaatttctt tttttacttt ctatttttaa tttatatatt
      781 tatattaaaa aatttaaatt ataattattt ttatagcacg tgatgaaaag gacccaggtg
      841 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      901 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      961 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1021 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1081 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1141 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1201 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1261 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1321 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1381 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1441 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1501 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1561 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1621 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1681 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1741 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1801 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1861 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1921 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1981 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2041 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2101 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2161 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2221 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2281 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2341 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2401 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2461 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2521 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2581 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2641 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2701 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2761 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     2821 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2881 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2941 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     3001 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctccggatg caagggttcg
     3061 aatcccttag ctctcattat tttttgcttt ttctcttgag gtcacatgat cgcaaaatgg
     3121 caaatggcac gtgaagctgt cgatattggg gaactgtggt ggttggcaaa tgactaatta
     3181 agttagtcaa ggcgccatcc tcatgaaaac tgtgtaacat aataaccgaa gtgtcgaaaa
     3241 ggtggcacct tgtccaattg aacacgctcg atgaaaaaaa taagatatat ataaggttaa
     3301 gtaaagcgtc tgttagaaag gaagtttttc ctttttcttg ctctcttgtc ttttcatcta
     3361 ctatttcctt cgtgtaatac agggtcgtca gatacataga tacaattcta ttacccccat
     3421 ccatactcta gaactagtgg atccaccatg gtgctgttgg cagcagcagt ctgtacaaaa
     3481 gcaggaaaag ctattgtttc tcgacagttt gtagagatga ctcgaactcg gattgagggc
     3541 ttgttagcag cttttccaaa gctcatgaac actggaaaac aacatacatt tgtggaaaca
     3601 gagagtgtca gatatgttta ccagcctatg gagaaattgt acatggtact gatcaccacc
     3661 aaaaacagca acatcttaga agatctggag accctaaggc tcttctcaag agtgattcct
     3721 gaatattgcc gagccttaga agagaatgaa atatctgagc actgttttga tttgattttt
     3781 gcatttgacg aaattgttgc cttgggacat caccatcacc atcactgacc ggaattcgat
     3841 atcaagctta tcgataccgt cgacctcgag tcatgtaatt agttatgtca cgcttacatt
     3901 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
     3961 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa
     4021 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
     4081 ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta
     4141 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     4201 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     4261 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4321 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4381 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4441 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4501 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4561 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4621 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4681 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4741 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     4801 cgcatctgtg cggtatttca caccgcatat cgaccggtcg aggagaactt ctagtatatc
     4861 tacataccta atattattgc cttattaaaa atggaatccc aacaattaca tcaaaatcca
     4921 cattctcttc aaaatcaatt gtcctgtact tccttgttca tgtgtgttca aaaacgttat
     4981 atttatagga taattatact ctatttctca acaagtaatt ggttgtttgg ccgagcggtc
     5041 taaggcgcct gattcaagaa atatcttgac cgcagttaac tgtgggaata ctcaggtatc
     5101 gtaagatgca agagttcgaa tctcttagca accattattt ttttcctcaa cataacgaga
     5161 acacacaggg gcgctatcgc acagaatcaa attcgatgac tggaaatttt ttgttaattt
     5221 cagaggtcgc ctgacgcata tacctttttc aactgaaaaa ttgggagaaa aaggaaaggt
     5281 gagagcgccg gaaccggctt ttcatataga atagagaagc gttcatgact aaatgcttgc
     5341 atcacaatac ttgaagttga caatattatt taaggaccta ttgttttttc caataggtgg
     5401 ttagcaatcg tcttactttc taacttttct taccttttac atttcagcaa tatatatata
     5461 tatatttcaa ggatatacca ttctaatgtc tgcccctaag aagatcgtcg ttttgccagg
     5521 tgaccacgtt ggtcaagaaa tcacagccga agccattaag gttcttaaag ctatttctga
     5581 tgttcgttcc aatgtcaagt tcgatttcga aaatcattta attggtggtg ctgctatcga
     5641 tgctacaggt gttccacttc cagatgaggc gctggaagcc tccaagaagg ctgatgccgt
     5701 tttgttaggt gctgtgggtg gtcctaaatg gggtaccggt agtgttagac ctgaacaagg
     5761 tttactaaaa atccgtaaag aacttcaatt gtacgccaac ttaagaccat gtaactttgc
     5821 atccgactct cttttagact tatctccaat caagccacaa tttgctaaag gtactgactt
     5881 cgttgttgtc agagaattag tgggaggtat ttactttggt aagagaaagg aagacgatgg
     5941 tgatggtgtc gcttgggata gtgaacaata caccgttcca gaagtgcaaa gaatcacaag
     6001 aatggccgct ttcatggccc tacaacatga gccaccattg cctatttggt ccttggataa
     6061 agctaatgtt ttggcctctt caagattatg gagaaaaact gtggaggaaa ccatcaagaa
     6121 cgaattccct acattgaagg ttcaacatca attgattgat tctgccgcca tgatcctagt
     6181 taagaaccca acccacctaa atggtattat aatcaccagc aacatgtttg gtgatatcat
     6241 ctccgatgaa gcctccgtta tcccaggttc cttgggtttg ttgccatctg cgtccttggc
     6301 ctctttgcca gacaagaaca ccgcatttgg tttgtacgaa ccatgccacg gttctgctcc
     6361 agatttgcca aagaataagg tcaaccctat cgccactatc ttgtctgctg caatgatgtt
     6421 gaaattgtca ttgaacttgc ctgaagaagg taaggccatt gaagatgcag ttaaaaaggt
     6481 tttggatgca ggtatcagaa ctggtgattt aggtggttcc aacagtacca ccgaagtcgg
     6541 tgatgctgtc gccgaagaag ttaagaaaat ccttgcttaa aaagattctc tttttttatg
     6601 atatttgtac ataaacttta taaatgaaat tcataataga aacgacacga aattacaaaa
     6661 tggaatatgt tcatagggta gacgaaacta tatacgcaat ctacatacat ttatcaagaa
     6721 ggagaaaaag gaggatgtaa aggaatacag gtaagcaaat tgatactaat ggctcaacgt
     6781 gataaggaaa aagaattgca ctttaacatt aatattgaca aggaggaggg caccacacaa
     6841 aaagttaggt gtaacagaaa atcatgaaac tatgattcct aatttatata ttggaggatt
     6901 ttctctaaaa aaaaaaaaat acaacaaata aaaaacactc aatgacctga ccatttgatg
     6961 gagtttaagt caataccttc ttgaaccatt tc
//
