LOCUS       sequence-200159-        7163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ret2LD2-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 Ret2LD2-alpha
FEATURES             Location/Qualifiers
     source          1..7163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(99..118)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     218..240
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(278..296)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    338..841
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        878..982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(983..1051,1052..1843)
                     /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(1201..1220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2014..2602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2503..2522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2756..2773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2890..2911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2926..2931,2932..2949,2950..2956)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2964..2980
                     /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     2969..2991
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2988..3004
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2988..3004
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3023..3043
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3025..3043
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3094..3443
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3444..3460
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3444..3460
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3996..4013
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4040..4056)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4041..4057)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4059..4306
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4075..4093)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4324..4343)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4325..4343)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4353..4370)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4353..4369)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4362..4384)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4510..4965
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4597..4616)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4807..4828
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5266..5673
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5674..6768
                     /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 ccttcttgaa ccatttccca taatggtgaa agttccctca agaattttac tctgtcagaa
       61 acggccttaa cgacgtagtc gatatggtgc actctcagta caatctgctc tgatgccgca
      121 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
      181 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
      241 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
      301 taggttaatg tcatgataat aatggtttct taggacggat cgcttgcctg taacttacac
      361 gcgcctcgta tcttttaatg atggaataat ttgggaattt actctgtgtt tatttatttt
      421 tatgttttgt atttggattt tagaaagtaa ataaagaagg tagaagagtt acggaatgaa
      481 gaaaaaaaaa taaacaaagg tttaaaaaat ttcaacaaaa agcgtacttt acatatatat
      541 ttattagaca agaaaagcag attaaataga tatacattcg attaacgata agtaaaatgt
      601 aaaatcacag gattttcgtg tgtggtcttc tacacagaca agatgaaaca attcggcatt
      661 aatacctgag agcaggaaga gcaagataaa aggtagtatt tgttggcgat ccccctagag
      721 tcttttacat cttcggaaaa caaaaactat tttttcttta atttcttttt ttactttcta
      781 tttttaattt atatatttat attaaaaaat ttaaattata attattttta tagcacgtga
      841 tgaaaaggac ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      901 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      961 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1021 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1081 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1141 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     1201 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     1261 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1321 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1381 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     1441 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1501 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1561 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1621 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1681 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1741 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1801 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1861 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1921 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1981 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2041 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2101 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2161 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2221 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2281 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2341 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2401 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2461 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2521 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2581 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2641 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2701 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2761 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     2821 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     2881 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     2941 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     3001 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct
     3061 ccggatgcaa gggttcgaat cccttagctc tcattatttt ttgctttttc tcttgaggtc
     3121 acatgatcgc aaaatggcaa atggcacgtg aagctgtcga tattggggaa ctgtggtggt
     3181 tggcaaatga ctaattaagt tagtcaaggc gccatcctca tgaaaactgt gtaacataat
     3241 aaccgaagtg tcgaaaaggt ggcaccttgt ccaattgaac acgctcgatg aaaaaaataa
     3301 gatatatata aggttaagta aagcgtctgt tagaaaggaa gtttttcctt tttcttgctc
     3361 tcttgtcttt tcatctacta tttccttcgt gtaatacagg gtcgtcagat acatagatac
     3421 aattctatta cccccatcca tactctagaa ctagtggatc catggtggtt ttggctgctt
     3481 ctatcactac gcgtcagggg aaacctctct tgtccaggca attcaaagat ctgtccaagg
     3541 atcgagtcct agagttgctg tccaatttcc agaacctagt ctccgagatc tcgtcagacc
     3601 acacatttgt cgaggacaag cacgtccgct acgtgtatag gccctttgat aactactaca
     3661 ttattcttat cacaaaccgc cagtccaata ttatcaagga tttggccaca ttaaatctgt
     3721 tttcacagac gatcaactcc tacttgtcta gtttccaaga ccaagagatt tttcacaatg
     3781 cctttgagat cttgtcctcg ttcgacgaaa tcgtgtcaat gggcgggtac aaagagaact
     3841 tgtcctttac tcaagtgcaa acatacttgt ctatggagtc gcacgaagaa agaatccagg
     3901 aaattattga gcgtaataaa gagatcgaag ctactgagga gaggaagcgc cgtgctaagg
     3961 aaattgctag aaaagaacat gaaaggaaac acgggcatca ccatcaccat cactgaccgg
     4021 aattcgatat caagcttatc gataccgtcg acctcgagtc atgtaattag ttatgtcacg
     4081 cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     4141 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat
     4201 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa
     4261 aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcggcc ggtacccaat
     4321 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     4381 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     4441 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     4501 ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4561 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4621 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4681 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4741 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4801 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4861 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4921 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     4981 tctccttacg catctgtgcg gtatttcaca ccgcatatcg accggtcgag gagaacttct
     5041 agtatatcta catacctaat attattgcct tattaaaaat ggaatcccaa caattacatc
     5101 aaaatccaca ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa
     5161 aacgttatat ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc
     5221 gagcggtcta aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact
     5281 caggtatcgt aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca
     5341 taacgagaac acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt
     5401 gttaatttca gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa
     5461 ggaaaggtga gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa
     5521 atgcttgcat cacaatactt gaagttgaca atattattta aggacctatt gttttttcca
     5581 ataggtggtt agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata
     5641 tatatatata tatttcaagg atataccatt ctaatgtctg cccctaagaa gatcgtcgtt
     5701 ttgccaggtg accacgttgg tcaagaaatc acagccgaag ccattaaggt tcttaaagct
     5761 atttctgatg ttcgttccaa tgtcaagttc gatttcgaaa atcatttaat tggtggtgct
     5821 gctatcgatg ctacaggtgt tccacttcca gatgaggcgc tggaagcctc caagaaggct
     5881 gatgccgttt tgttaggtgc tgtgggtggt cctaaatggg gtaccggtag tgttagacct
     5941 gaacaaggtt tactaaaaat ccgtaaagaa cttcaattgt acgccaactt aagaccatgt
     6001 aactttgcat ccgactctct tttagactta tctccaatca agccacaatt tgctaaaggt
     6061 actgacttcg ttgttgtcag agaattagtg ggaggtattt actttggtaa gagaaaggaa
     6121 gacgatggtg atggtgtcgc ttgggatagt gaacaataca ccgttccaga agtgcaaaga
     6181 atcacaagaa tggccgcttt catggcccta caacatgagc caccattgcc tatttggtcc
     6241 ttggataaag ctaatgtttt ggcctcttca agattatgga gaaaaactgt ggaggaaacc
     6301 atcaagaacg aattccctac attgaaggtt caacatcaat tgattgattc tgccgccatg
     6361 atcctagtta agaacccaac ccacctaaat ggtattataa tcaccagcaa catgtttggt
     6421 gatatcatct ccgatgaagc ctccgttatc ccaggttcct tgggtttgtt gccatctgcg
     6481 tccttggcct ctttgccaga caagaacacc gcatttggtt tgtacgaacc atgccacggt
     6541 tctgctccag atttgccaaa gaataaggtc aaccctatcg ccactatctt gtctgctgca
     6601 atgatgttga aattgtcatt gaacttgcct gaagaaggta aggccattga agatgcagtt
     6661 aaaaaggttt tggatgcagg tatcagaact ggtgatttag gtggttccaa cagtaccacc
     6721 gaagtcggtg atgctgtcgc cgaagaagtt aagaaaatcc ttgcttaaaa agattctctt
     6781 tttttatgat atttgtacat aaactttata aatgaaattc ataatagaaa cgacacgaaa
     6841 ttacaaaatg gaatatgttc atagggtaga cgaaactata tacgcaatct acatacattt
     6901 atcaagaagg agaaaaagga ggatgtaaag gaatacaggt aagcaaattg atactaatgg
     6961 ctcaacgtga taaggaaaaa gaattgcact ttaacattaa tattgacaag gaggagggca
     7021 ccacacaaaa agttaggtgt aacagaaaat catgaaacta tgattcctaa tttatatatt
     7081 ggaggatttt ctctaaaaaa aaaaaaatac aacaaataaa aaacactcaa tgacctgacc
     7141 atttgatgga gtttaagtca ata
//
