LOCUS       sequence-200164-        6992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ret2LD - 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 Ret2LD
FEATURES             Location/Qualifiers
     source          1..6992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(192..211)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     311..333
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(371..389)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    431..934
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        971..1075
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1076..1144,1145..1936)
                     /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(1294..1313)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2107..2695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2596..2615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2849..2866
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2983..3004
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3019..3024,3025..3042,3043..3049)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3057..3073
                     /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     3062..3084
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3081..3097
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3081..3097
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3116..3136
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3118..3136
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3187..3536
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     3537..3553
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3537..3553
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3918..3935
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3962..3978)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3963..3979)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      3981..4228
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3997..4015)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4246..4265)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4247..4265)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4275..4292)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4275..4291)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4284..4306)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4432..4887
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4519..4538)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4729..4750
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5188..5595
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5596..6690
                     /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 tttatatatt ggaggatttt ctctaaaaaa aaaaaaatac aacaaataaa aaacactcaa
       61 tgacctgacc atttgatgga gtttaagtca ataccttctt gaaccatttc ccataatggt
      121 gaaagttccc tcaagaattt tactctgtca gaaacggcct taacgacgta gtcgatatgg
      181 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
      241 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
      301 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
      361 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
      421 tcttaggacg gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat
      481 aatttgggaa tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag
      541 taaataaaga aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa
      601 aatttcaaca aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat
      661 agatatacat tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc
      721 ttctacacag acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat
      781 aaaaggtagt atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac
      841 tattttttct ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa
      901 aatttaaatt ataattattt ttatagcacg tgatgaaaag gacccaggtg gcacttttcg
      961 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1021 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1081 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1141 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1201 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1261 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     1321 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1381 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1441 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1501 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1561 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     1621 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1681 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1741 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     1801 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1861 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     1921 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     1981 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2041 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2101 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2161 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2221 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2281 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2341 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2401 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2461 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2521 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2581 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2641 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2701 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2761 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2821 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     2881 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     2941 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     3001 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     3061 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     3121 taaccctcac taaagggaac aaaagctgga gctccggatg caagggttcg aatcccttag
     3181 ctctcattat tttttgcttt ttctcttgag gtcacatgat cgcaaaatgg caaatggcac
     3241 gtgaagctgt cgatattggg gaactgtggt ggttggcaaa tgactaatta agttagtcaa
     3301 ggcgccatcc tcatgaaaac tgtgtaacat aataaccgaa gtgtcgaaaa ggtggcacct
     3361 tgtccaattg aacacgctcg atgaaaaaaa taagatatat ataaggttaa gtaaagcgtc
     3421 tgttagaaag gaagtttttc ctttttcttg ctctcttgtc ttttcatcta ctatttcctt
     3481 cgtgtaatac agggtcgtca gatacataga tacaattcta ttacccccat ccatactcta
     3541 gaactagtgg atccatggtg gttttggctg cttctatcac tacgcgtcag gggaaacctc
     3601 tcttgtccag gcaattcaaa gatctgtcca aggatcgagt cctagagttg ctgtccaatt
     3661 tccagaacct agtctccgag atctcgtcag accacacatt tgtcgaggac aagcacgtcc
     3721 gctacgtgta taggcccttt gataactact acattattct tatcacaaac cgccagtcca
     3781 atattatcaa ggatttggcc acattaaatc tgttttcaca gacgatcaac tcctacttgt
     3841 ctagtttcca agaccaagag atttttcaca atgcctttga gatcttgtcc tcgttcgacg
     3901 aaatcgtgtc aatgggccat caccatcacc atcactgacc ggaattcgat atcaagctta
     3961 tcgataccgt cgacctcgag tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     4021 ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     4081 ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt
     4141 ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt
     4201 tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta tagtgagtcg
     4261 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     4321 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     4381 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     4441 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4501 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4561 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4621 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4681 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4741 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4801 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4861 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     4921 cggtatttca caccgcatat cgaccggtcg aggagaactt ctagtatatc tacataccta
     4981 atattattgc cttattaaaa atggaatccc aacaattaca tcaaaatcca cattctcttc
     5041 aaaatcaatt gtcctgtact tccttgttca tgtgtgttca aaaacgttat atttatagga
     5101 taattatact ctatttctca acaagtaatt ggttgtttgg ccgagcggtc taaggcgcct
     5161 gattcaagaa atatcttgac cgcagttaac tgtgggaata ctcaggtatc gtaagatgca
     5221 agagttcgaa tctcttagca accattattt ttttcctcaa cataacgaga acacacaggg
     5281 gcgctatcgc acagaatcaa attcgatgac tggaaatttt ttgttaattt cagaggtcgc
     5341 ctgacgcata tacctttttc aactgaaaaa ttgggagaaa aaggaaaggt gagagcgccg
     5401 gaaccggctt ttcatataga atagagaagc gttcatgact aaatgcttgc atcacaatac
     5461 ttgaagttga caatattatt taaggaccta ttgttttttc caataggtgg ttagcaatcg
     5521 tcttactttc taacttttct taccttttac atttcagcaa tatatatata tatatttcaa
     5581 ggatatacca ttctaatgtc tgcccctaag aagatcgtcg ttttgccagg tgaccacgtt
     5641 ggtcaagaaa tcacagccga agccattaag gttcttaaag ctatttctga tgttcgttcc
     5701 aatgtcaagt tcgatttcga aaatcattta attggtggtg ctgctatcga tgctacaggt
     5761 gttccacttc cagatgaggc gctggaagcc tccaagaagg ctgatgccgt tttgttaggt
     5821 gctgtgggtg gtcctaaatg gggtaccggt agtgttagac ctgaacaagg tttactaaaa
     5881 atccgtaaag aacttcaatt gtacgccaac ttaagaccat gtaactttgc atccgactct
     5941 cttttagact tatctccaat caagccacaa tttgctaaag gtactgactt cgttgttgtc
     6001 agagaattag tgggaggtat ttactttggt aagagaaagg aagacgatgg tgatggtgtc
     6061 gcttgggata gtgaacaata caccgttcca gaagtgcaaa gaatcacaag aatggccgct
     6121 ttcatggccc tacaacatga gccaccattg cctatttggt ccttggataa agctaatgtt
     6181 ttggcctctt caagattatg gagaaaaact gtggaggaaa ccatcaagaa cgaattccct
     6241 acattgaagg ttcaacatca attgattgat tctgccgcca tgatcctagt taagaaccca
     6301 acccacctaa atggtattat aatcaccagc aacatgtttg gtgatatcat ctccgatgaa
     6361 gcctccgtta tcccaggttc cttgggtttg ttgccatctg cgtccttggc ctctttgcca
     6421 gacaagaaca ccgcatttgg tttgtacgaa ccatgccacg gttctgctcc agatttgcca
     6481 aagaataagg tcaaccctat cgccactatc ttgtctgctg caatgatgtt gaaattgtca
     6541 ttgaacttgc ctgaagaagg taaggccatt gaagatgcag ttaaaaaggt tttggatgca
     6601 ggtatcagaa ctggtgattt aggtggttcc aacagtacca ccgaagtcgg tgatgctgtc
     6661 gccgaagaag ttaagaaaat ccttgcttaa aaagattctc tttttttatg atatttgtac
     6721 ataaacttta taaatgaaat tcataataga aacgacacga aattacaaaa tggaatatgt
     6781 tcatagggta gacgaaacta tatacgcaat ctacatacat ttatcaagaa ggagaaaaag
     6841 gaggatgtaa aggaatacag gtaagcaaat tgatactaat ggctcaacgt gataaggaaa
     6901 aagaattgca ctttaacatt aatattgaca aggaggaggg caccacacaa aaagttaggt
     6961 gtaacagaaa atcatgaaac tatgattcct aa
//
