LOCUS       sequence-131718-        8242 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Histidine selection; Used to insert a GAL1 promoter and 24 
            repetitive PP7 version 3 RNA stem-loops before GLT1's start codon 
            (5' tagging).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8242)
  AUTHORS   Saroufim MA, Bensidoun P, Raymond P, Rahman S, Krause MR, Oeffinger 
            M, Zenklusen D
  TITLE     The nuclear basket mediates perinuclear mRNA scanning in budding 
            yeast.
  JOURNAL   J Cell Biol. 2015 Dec 21;211(6):1131-40. doi: 10.1083/jcb.201503070.
  PUBMED    26694838
REFERENCE   2  (bases 1 to 8242)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2015-12-21- 21"; volume: "211"; 
            issue: "6"; pages: "1131-40"
COMMENT     Sequence Label: pDZ305 GTL1 GAL1p 24xPP7 V3 <HIS>
FEATURES             Location/Qualifiers
     source          1..8242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        416..759
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             760..1458
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1462..1659
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        2043..2484
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2043..2160
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2455..2478
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(4317..4336)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4318..4336)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4343..4360)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4343..4359)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4352..4374)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4497..4799
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4678..4697
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5148..5942
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5148..5942
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5202..5221)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5812..5831
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(6488..6866)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     6813..6832
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7174..7762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7663..7682
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7916..7933
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8050..8071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8086..8091,8092..8109,8110..8116)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8124..8140
                     /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     8129..8151
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8148..8164
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8148..8164
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8185..8203
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaattaaac cgttcaaccc tatacagcgc aaatttggtc ctaatacact tcggttttaa
       61 tgcgtcaatc cgattggctc cgataagctt ttgcacattt ttcaagtcat atgtcacgac
      121 gaacgtgatg cccgcaaaac gtaaaaaaaa gaagcaataa attgcccaaa aaaaaaatat
      181 aaactgcttt tcttagtcat tcataagttc tgatggtctt ccttatctta ttgtattttt
      241 cgtcctattg tttcatttct taccaaatta atcaattctt atatcttact tgataacaca
      301 ccaaactaat cgtctccaca tcataggaag ataggaaatt gctatctcag tcctatacta
      361 cgcagacctc gagtctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat
      421 ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga
      481 ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt
      541 tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg
      601 aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat
      661 aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct
      721 aggatacagt tctcacatca catccgaaca taaacaacca tggcagaacc agcccaaaaa
      781 aagcaaaaac aaactgttca ggagcgcaag gcgtttatct cccgtatcac taatgaaact
      841 aaaattcaaa tcgctatttc gctgaatggt ggttatattc aaataaaaga ttcgattctt
      901 cctgcaaaga aggatgacga tgtagcttcc caagctactc agtcacaggt catcgatatt
      961 cacacaggtg ttggcttttt ggatcatatg atccatgcgt tggcaaaaca ctctggttgg
     1021 tctcttattg ttgaatgtat tggtgacctg cacattgacg atcaccatac taccgaagat
     1081 tgcggtatcg cattagggca agcgttcaaa gaagcaatgg gtgctgtccg tggtgtaaaa
     1141 agattcggta ctgggttcgc accattggat gaggcgctat cacgtgccgt agtcgattta
     1201 tctagtagac catttgctgt aatcgacctt ggattgaaga gagagatgat tggtgattta
     1261 tccactgaaa tgattccaca ctttttggaa agtttcgcgg aggcggccag aattactttg
     1321 catgttgatt gtctgagagg tttcaacgat caccacagaa gtgagagtgc gttcaaggct
     1381 ttggctgttg ccataagaga agctatttct agcaatggca ccaatgacgt tccctcaacc
     1441 aaaggtgttt tgatgtgaag tactgacaat aaaaagattc ttgttttcaa gaacttgtca
     1501 tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat
     1561 attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata
     1621 tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc
     1681 cgccatccag agctccaggt tatcagcaac aacacagtca tatccattct caattagctc
     1741 taccacagtg tgtgaaccaa tgtatccagc accacctgta accaaaacaa ttttagaagt
     1801 actttcactt tgtaactgag ctgtcattta tattgaattt tcaaaaattc ttactttttt
     1861 tttggatgga cgcaaagaag tttaataatc atattacatg gcattaccac catatacata
     1921 tccatatcta atcttactta tatgttgtgg aaatgtaaag agccccatta tcttagccta
     1981 aaaaaacctt ctctttggaa ctttcagtaa tacgcttaac tgctcattgc tatattgaag
     2041 tacggattag aagccgccga gcgggcgaca gccctccgac ggaagactct cctccgtgcg
     2101 tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg cctcgcgccg cactgctccg
     2161 aacaataaag attctacaat actagctttt atggttatga agaggaaaaa ttggcagtaa
     2221 cctggcccca caaaccttca aattaacgaa tcaaattaac aaccatagga tgataatgcg
     2281 attagttttt tagccttatt tctggggtaa ttaatcagcg aagcgatgat ttttgatcta
     2341 ttaacagata tataaatgga aaagctgcat aaccacttta actaatactt tcaacatttt
     2401 cagtttgtat tacttcttat tcaaatgtca taaaagtatc aacaaaaaat tgttaatata
     2461 cctctatact ttaacgtcaa ggagggatcc taaggtacct aattgcctag aaaggagcag
     2521 acgatatggc gtcgctccct gcaggtcgac tctagaaacc agcagagcat atgggctcgc
     2581 tggctgcagt attcccgggt tcattagatc ctaaggtacc taattgccta gaaaggagca
     2641 gacgatatgg cgtcgctccc tgcaggtcga ctctagaaac cagcagagca tatgggctcg
     2701 ctggctgcag tattcccggg ttcattagat cctaaggtac ctaattgcct agaaaggagc
     2761 agacgatatg gcgtcgctcc ctgcaggtcg actctagaaa ccagcagagc atatgggctc
     2821 gctggctgca gtattcccgg gttcattaga tcctaaggta cctaattgcc tagaaaggag
     2881 cagacgatat ggcgtcgctc cctgcaggtc gactctagaa accagcagag catatgggct
     2941 cgctggctgc agtattcccg ggttcattag atcctaaggt acctaattgc ctagaaagga
     3001 gcagacgata tggcgtcgct ccctgcaggt cgactctaga aaccagcaga gcatatgggc
     3061 tcgctggctg cagtattccc gggttcatta gatcctaagg tacctaattg cctagaaagg
     3121 agcagacgat atggcgtcgc tccctgcagg tcgactctag aaaccagcag agcatatggg
     3181 ctcgctggct gcagtattcc cgggttcatt agatcctaag gtacctaatt gcctagaaag
     3241 gagcagacga tatggcgtcg ctccctgcag gtcgactcta gaaaccagca gagcatatgg
     3301 gctcgctggc tgcagtattc ccgggttcat tagatcctaa ggtacctaat tgcctagaaa
     3361 ggagcagacg atatggcgtc gctccctgca ggtcgactct agaaaccagc agagcatatg
     3421 ggctcgctgg ctgcagtatt cccgggttca ttagatccta aggtacctaa ttgcctagaa
     3481 aggagcagac gatatggcgt cgctccctgc aggtcgactc tagaaaccag cagagcatat
     3541 gggctcgctg gctgcagtat tcccgggttc attagatcct aaggtaccta attgcctaga
     3601 aaggagcaga cgatatggcg tcgctccctg caggtcgact ctagaaacca gcagagcata
     3661 tgggctcgct ggctgcagta ttcccgggtt cattagatcc taaggtacct aattgcctag
     3721 aaaggagcag acgatatggc gtcgctccct gcaggtcgac tctagaaacc agcagagcat
     3781 atgggctcgc tggctgcagt attcccgggt tcattagatc ctaaggtacc taattgccta
     3841 gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga ctctagaaac cagcagagca
     3901 tatgggctcg ctggctgcag tattcccggg ttcattagat ccatgtccca ggatagaatt
     3961 ttgttagata gatctatgcc agtgttgaaa tcagacaatt tcgatccatt ggaagaagct
     4021 tacgaaggtg ggacaattca aaactataac gatgaacacc atcttcataa atcttgggca
     4081 aatgtgattc cggacaaacg aggactttac gaccctgatt atgaacatga cgcttgtggt
     4141 gtcggtttcg tagcaaataa gcatggtgaa cagtctcaca agattgttac tgacgctaga
     4201 tatcttttag tgaatatgac acatcgtggt gccgtctcat ctgatgggaa cggtgacggt
     4261 gccggtattc tgctaggtat cccaagggcg aattcgcggc cgctaaattc aattcgccct
     4321 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     4381 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     4441 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctatac gtacggcagt
     4501 ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg
     4561 atattattga cacgccgggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt
     4621 cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
     4681 tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc
     4741 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa
     4801 tgtcaggcat gagattatca aaaaggatct tcacctagat ccttttcacg tagaaagcca
     4861 gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg
     4921 aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag
     4981 actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta
     5041 aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc
     5101 gcaggggatc aagctctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5161 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5221 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5281 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     5341 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5401 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5461 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5521 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5581 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5641 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     5701 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5761 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5821 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5881 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5941 gaattattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     6001 tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6061 tttttctaaa tacattcaaa tatgtatccg ctcatgagat tatcaaaaag gatcttcacc
     6121 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6181 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6241 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6301 ccatctggcc ccagtgctgc aatgataccg cgagaccacg ctcaccggct ccagatttat
     6361 cagcaataaa ccagccagcc ggaaggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6421 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6481 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6541 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6601 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6661 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6721 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6781 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6841 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6901 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6961 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7021 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7081 ttatcagggt tattgtctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7141 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7201 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7261 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7321 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7381 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7441 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7501 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7561 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7621 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7681 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7741 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7801 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7861 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     7921 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     7981 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     8041 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     8101 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     8161 tgaccatgat tacgccaagc tcagaattaa ccctcactaa agggactagt cctgcaggtt
     8221 taaacgaatt cgcccttata ta
//
