LOCUS       sequence-200162-        7163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  delta-CLD2-alpha - His tagged (uHD deleted).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7163)
  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 7163)
  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-CLD2-alpha
FEATURES             Location/Qualifiers
     source          1..7163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(79..98)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     198..220
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(258..276)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    318..821
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        858..962
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(963..1031,1032..1823)
                     /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(1181..1200)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1994..2582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2483..2502
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2736..2753
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2870..2891
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2906..2911,2912..2929,2930..2936)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2944..2960
                     /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     2949..2971
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2968..2984
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2968..2984
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3003..3023
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3005..3023
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3074..3423
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3424..3440
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3424..3440
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3976..3993
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4020..4036)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4021..4037)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4039..4286
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4055..4073)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4304..4323)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4305..4323)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4333..4350)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4333..4349)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4342..4364)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4490..4945
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4577..4596)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4787..4808
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5246..5653
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5654..6748
                     /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 taatggtgaa agttccctca agaattttac tctgtcagaa acggccttaa cgacgtagtc
       61 gatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
      121 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
      181 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
      241 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
      301 aatggtttct taggacggat cgcttgcctg taacttacac gcgcctcgta tcttttaatg
      361 atggaataat ttgggaattt actctgtgtt tatttatttt tatgttttgt atttggattt
      421 tagaaagtaa ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg
      481 tttaaaaaat ttcaacaaaa agcgtacttt acatatatat ttattagaca agaaaagcag
      541 attaaataga tatacattcg attaacgata agtaaaatgt aaaatcacag gattttcgtg
      601 tgtggtcttc tacacagaca agatgaaaca attcggcatt aatacctgag agcaggaaga
      661 gcaagataaa aggtagtatt tgttggcgat ccccctagag tcttttacat cttcggaaaa
      721 caaaaactat tttttcttta atttcttttt ttactttcta tttttaattt atatatttat
      781 attaaaaaat ttaaattata attattttta tagcacgtga tgaaaaggac ccaggtggca
      841 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      901 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      961 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1021 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1081 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1141 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1201 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1261 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1321 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1381 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1441 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1501 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1561 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1621 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1681 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1741 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     1801 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     1861 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1921 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1981 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2041 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2101 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2161 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2221 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2281 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2341 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2401 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2461 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2521 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2581 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2641 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2701 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     2761 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     2821 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     2881 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     2941 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     3001 gcgcaattaa ccctcactaa agggaacaaa agctggagct ccggatgcaa gggttcgaat
     3061 cccttagctc tcattatttt ttgctttttc tcttgaggtc acatgatcgc aaaatggcaa
     3121 atggcacgtg aagctgtcga tattggggaa ctgtggtggt tggcaaatga ctaattaagt
     3181 tagtcaaggc gccatcctca tgaaaactgt gtaacataat aaccgaagtg tcgaaaaggt
     3241 ggcaccttgt ccaattgaac acgctcgatg aaaaaaataa gatatatata aggttaagta
     3301 aagcgtctgt tagaaaggaa gtttttcctt tttcttgctc tcttgtcttt tcatctacta
     3361 tttccttcgt gtaatacagg gtcgtcagat acatagatac aattctatta cccccatcca
     3421 tactctagaa ctagtggatc caccatggtg ctgttggcag cagcagtctg tacaaaagca
     3481 ggaaaagcta ttgtttctcg acagtttgta gagatgactc gaactcggat tgagggcttg
     3541 ttagcagctt ttccaaagct catgaacact ggaaaacaac atacatttgt ggaaacagag
     3601 agtgtcagat atgtttacca gcctatggag aaattgtaca tggtactgat caccaccaaa
     3661 aacagcaaca tcttagaaga tctggagacc ctaaggctct tctcaagagt gattcctgaa
     3721 tattgccgag ccttagaaga gaatgaaata tctgagcact gttttgattt gatttttgca
     3781 tttgacgaaa ttgttgcctt gggataccgg gagaatgtta acctggcaca gatcagaacc
     3841 ttcacagaaa tggattccca tgaggagaag gtgttcagag cagtcagaga gactcaagag
     3901 cgtgaagcca aggctgagat gcggcgtaaa gcaaaagaat tacagcaggc ccgaagagat
     3961 gcagagagac agggccatca ccatcaccat cactgaccgg aattcgatat caagcttatc
     4021 gataccgtcg acctcgagtc atgtaattag ttatgtcacg cttacattca cgccctcccc
     4081 ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
     4141 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     4201 ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     4261 tgggacgctc gaaggcttta atttgcggcc ggtacccaat tcgccctata gtgagtcgta
     4321 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     4381 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     4441 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt
     4501 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4561 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4621 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4681 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4741 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4801 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4861 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4921 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     4981 gtatttcaca ccgcatatcg accggtcgag gagaacttct agtatatcta catacctaat
     5041 attattgcct tattaaaaat ggaatcccaa caattacatc aaaatccaca ttctcttcaa
     5101 aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa aacgttatat ttataggata
     5161 attatactct atttctcaac aagtaattgg ttgtttggcc gagcggtcta aggcgcctga
     5221 ttcaagaaat atcttgaccg cagttaactg tgggaatact caggtatcgt aagatgcaag
     5281 agttcgaatc tcttagcaac cattattttt ttcctcaaca taacgagaac acacaggggc
     5341 gctatcgcac agaatcaaat tcgatgactg gaaatttttt gttaatttca gaggtcgcct
     5401 gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga gagcgccgga
     5461 accggctttt catatagaat agagaagcgt tcatgactaa atgcttgcat cacaatactt
     5521 gaagttgaca atattattta aggacctatt gttttttcca ataggtggtt agcaatcgtc
     5581 ttactttcta acttttctta ccttttacat ttcagcaata tatatatata tatttcaagg
     5641 atataccatt ctaatgtctg cccctaagaa gatcgtcgtt ttgccaggtg accacgttgg
     5701 tcaagaaatc acagccgaag ccattaaggt tcttaaagct atttctgatg ttcgttccaa
     5761 tgtcaagttc gatttcgaaa atcatttaat tggtggtgct gctatcgatg ctacaggtgt
     5821 tccacttcca gatgaggcgc tggaagcctc caagaaggct gatgccgttt tgttaggtgc
     5881 tgtgggtggt cctaaatggg gtaccggtag tgttagacct gaacaaggtt tactaaaaat
     5941 ccgtaaagaa cttcaattgt acgccaactt aagaccatgt aactttgcat ccgactctct
     6001 tttagactta tctccaatca agccacaatt tgctaaaggt actgacttcg ttgttgtcag
     6061 agaattagtg ggaggtattt actttggtaa gagaaaggaa gacgatggtg atggtgtcgc
     6121 ttgggatagt gaacaataca ccgttccaga agtgcaaaga atcacaagaa tggccgcttt
     6181 catggcccta caacatgagc caccattgcc tatttggtcc ttggataaag ctaatgtttt
     6241 ggcctcttca agattatgga gaaaaactgt ggaggaaacc atcaagaacg aattccctac
     6301 attgaaggtt caacatcaat tgattgattc tgccgccatg atcctagtta agaacccaac
     6361 ccacctaaat ggtattataa tcaccagcaa catgtttggt gatatcatct ccgatgaagc
     6421 ctccgttatc ccaggttcct tgggtttgtt gccatctgcg tccttggcct ctttgccaga
     6481 caagaacacc gcatttggtt tgtacgaacc atgccacggt tctgctccag atttgccaaa
     6541 gaataaggtc aaccctatcg ccactatctt gtctgctgca atgatgttga aattgtcatt
     6601 gaacttgcct gaagaaggta aggccattga agatgcagtt aaaaaggttt tggatgcagg
     6661 tatcagaact ggtgatttag gtggttccaa cagtaccacc gaagtcggtg atgctgtcgc
     6721 cgaagaagtt aagaaaatcc ttgcttaaaa agattctctt tttttatgat atttgtacat
     6781 aaactttata aatgaaattc ataatagaaa cgacacgaaa ttacaaaatg gaatatgttc
     6841 atagggtaga cgaaactata tacgcaatct acatacattt atcaagaagg agaaaaagga
     6901 ggatgtaaag gaatacaggt aagcaaattg atactaatgg ctcaacgtga taaggaaaaa
     6961 gaattgcact ttaacattaa tattgacaag gaggagggca ccacacaaaa agttaggtgt
     7021 aacagaaaat catgaaacta tgattcctaa tttatatatt ggaggatttt ctctaaaaaa
     7081 aaaaaaatac aacaaataaa aaacactcaa tgacctgacc atttgatgga gtttaagtca
     7141 ataccttctt gaaccatttc cca
//
