LOCUS       sequence-267401-        7197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control for measuring background levels of yTRAP reporter protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7197)
  AUTHORS   Newby GA, Kiriakov S, Hallacli E, Kayatekin C, Tsvetkov P, Mancuso 
            CP, Bonner JM, Hesse WR, Chakrabortee S, Manogaran AL, Liebman SW, 
            Lindquist S, Khalil AS
  TITLE     A Genetic Tool to Track Protein Aggregates and Control Prion 
            Inheritance.
  JOURNAL   Cell. 2017 Nov 2;171(4):966-979.e18. doi: 
            10.1016/j.cell.2017.09.041. Epub 2017 Oct 19.
  PUBMED    29056345
REFERENCE   2  (bases 1 to 7197)
  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: 
            "10.1016/j.cell.2017.09.041"; journalName: "Cell"; date: "2017-11-2-
            2"; volume: "171"; issue: "4"; pages: "966-979.e18"
COMMENT     Sequence Label: pGAN213
FEATURES             Location/Qualifiers
     source          1..7197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     regulatory      1710..1719
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      2186..2373
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2241..2265)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            2448..3579
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     CDS             2798..3370
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      3382..3579
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     4142..4158
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4142..4158
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4172..4191)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4173..4191)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4198..4215)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4198..4214)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4207..4229)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4514..4533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4646..4667
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4821..4840)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4940..4962
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5000..5018)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5086..5190
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5191..5259,5260..6051)
                     /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(5409..5428)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6222..6810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6711..6730
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6964..6981
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7098..7119
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    7174..7190
                     /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     join(7179..7197,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca ctagtccgag gcctcgagat ccgatatcgc
       61 cgtggcggcc gccagctgaa gcttaattat cctgggcacg agtgaaacaa agctaaaacc
      121 tttatttagc atggccattg aatgtaacaa ttatatatat cgcaagcaca aaaaatcaag
      181 gagagagaac taccactttg ttcatgtgta caatgttcat tatctccata agcaaaaaaa
      241 aaaaaataga aaacatatgc tataaggttg atattctcac gagtaagcgg cacttgctac
      301 ttattgacat tgcagatttt tggctacaga aatagtatat tagagattat aattgctaat
      361 caaatcaaaa tataaaatta gtaaaccaaa ccatttatac ccttccttag tagttatgga
      421 ttgtttttta atgatatttc tgcaaaccaa agaaagattg ttatccagat agaatttagt
      481 tttgatattc atttttttgt tgaagattga acgccatatc tgggcctcat aattcaaaag
      541 acggtgccat tatcggtagc gtttcgcatt gtactggatt tcagaaattt cacagttgat
      601 gaatcgaaaa gaatggtctc attgcaacac gtaaggttaa gatgtccctt tttaccatta
      661 taggcaataa atgaatcata aaacgaccgt atactggtga aatagtaggg agaacgagta
      721 cctgtagtaa aaagtataaa tcatagttaa tcgggcaatg tccctcgatc aaggagtatt
      781 gtgtcatgtt cgagacaaac gccaacattt ttgtttcttt tggacaaatg ttgtttgcat
      841 ttatgatccg ttatattttg atctaatgta gagttgcacg tagttcttac tggcaaagaa
      901 atcgatgcat accaaaaaag aataaaggtg atatttgatc tttaccgttt agttccaacg
      961 taaaattgtg cctttggact taaaatggcg tcttaagagt gaggactcga aaatattaat
     1021 agagtgagga ctcgaaaata ttaatagagt gaggactcga aaatattaat agagtgagga
     1081 ctcgaaaata ttaatatcga tagagtgagg actcgaaaat attaatagag tgaggactcg
     1141 aaaatattaa tgaattcaga gtgaggactc gaaaatatta atagagtgag gactcgaaaa
     1201 tattaatggc cggcccagat ccgccaggcg tgtatatata gcgtggatgg ccaggcaact
     1261 ttagtgctga cacatacagg catatatata tgtgtgcgac gacacatgat catatggcat
     1321 gcatgtgctc tgtatgtata taaaactctt gttttcttct tttctctaaa tattctttcc
     1381 ttatacatta ggacctttgc agcataaatt actatacttc tatagacaca caaacacaaa
     1441 tacacacact gtttaaacaa aaaatgggcg aggaggataa catggcctct ctcccagcga
     1501 cacatgagtt acacatcttt ggctccatca acggtgtgga ctttgacatg gtgggtcagg
     1561 gcaccggcaa tccaaatgat ggttatgagg agttaaacct gaagtccacc aagggtgacc
     1621 tccagttctc cccctggatt ctggtccctc atatcgggta tggcttccat cagtacctgc
     1681 cctaccctga cgggatgtcg cctttccagg ccgccatggt agatggctcc ggataccaag
     1741 tccatcgcac aatgcagttt gaagatggtg cctcccttac tgttaactac cgctacacct
     1801 acgagggaag ccacatcaaa ggagaggccc aggtgaaggg gactggtttc cctgctgacg
     1861 gtcctgtgat gaccaactcg ctgaccgctg cggactggtg caggtcgaag aagacttacc
     1921 ccaacgacaa aaccatcatc agtaccttta agtggagtta caccactgga aatggcaagc
     1981 gctaccggag cactgcgcgg accacctaca cctttgccaa gccaatggcg gctaactatc
     2041 tgaagaacca gccgatgtac gtgttccgta agacggagct caagcactcc aagaccgagc
     2101 tcaacttcaa ggagtggcaa aaggccttta ccgatgtgat gggcatggac gagctgtaca
     2161 agtaaggcgc gccacttcta aataagcgaa tttcttatga tttatgattt ttattattaa
     2221 ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg
     2281 aaaattctta ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagta
     2341 tgaggtcgct cttattgacc acacctctac cggcagatcc gctagggata acagggtaat
     2401 atagatctgt ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc
     2461 agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc
     2521 cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc
     2581 gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc
     2641 ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt
     2701 aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca
     2761 gttctcacat cacatccgaa catcaattga aacaaccatg ggtaccactc ttgacgacac
     2821 ggcttaccgg taccgcacca gtgtcccggg ggacgccgag gccatcgagg cactggatgg
     2881 gtccttcacc accgacaccg tcttccgcgt caccgccacc ggggacggct tcaccctgcg
     2941 ggaggtgccg gtggacccgc ccctgaccaa ggtgttcccc gacgacgaat cggacgacga
     3001 atcggacgac ggggaggacg gcgacccgga ctcccggacg ttcgtcgcgt acggggacga
     3061 cggcgacctg gcgggcttcg tggtcgtctc gtactccggc tggaaccgcc ggctgaccgt
     3121 cgaggacatc gaggtcgccc cggagcaccg ggggcacggg gtcgggcgcg cgttgatggg
     3181 gctcgcgacg gagttcgccc gcgagcgggg cgccgggcac ctctggctgg aggtcaccaa
     3241 cgtcaacgca ccggcgatcc acgcgtaccg gcggatgggg ttcaccctct gcggcctgga
     3301 caccgccctg tacgacggca ccgcctcgga cggcgagcag gcgctctaca tgagcatgcc
     3361 ctgcccctaa agatcttcag tactgacaat aaaaagattc ttgttttcaa gaacttgtca
     3421 tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat
     3481 attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata
     3541 tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc
     3601 cgccatccag tgtcgaaaac gagctcgaat tcctggggga acaacttcac agaatgtttt
     3661 gtcatattgt cgaagtggtc acaaaacaag agaagttccg ccaattataa aaagggaacc
     3721 cgtatatttc agcttcacgg atgatttcca gggtgagagt actgtatatg ggcttacgat
     3781 agaaggccat aaaaatttct tgcttggcaa caaaatagaa gtgaaatcat gtcgaggctg
     3841 ctgtgtggga gaacagcata aaatatcaca aaaaaagaat ctaaaacact gtgttgcttg
     3901 tcccagaaag ggaatcaagt atttttataa agattggagt ggtaaaaatc gagtatgtgc
     3961 tagatgctat ggaagataca aattcagcgg tcatcactgt ataaattgca agtatgtacc
     4021 agaagcacgt gaagtgaaaa aggcaaaaga caaaggcgaa aaattgggca ttacgcccga
     4081 aggtttgcca gttaaaggac cagagtgtat aaaatgtggc ggaatcttac agtgcggccg
     4141 ctctagaact agtggatcga tccccaattc gccctatagt gagtcgtatt acaattcact
     4201 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     4261 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     4321 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     4381 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     4441 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4501 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4561 ccccgtcaag ctctaaatcg gggtgggcca tcgccctgat agacggtttt tcgccctttg
     4621 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4681 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     4741 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4801 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4861 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4921 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4981 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5041 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5101 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5161 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5221 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5281 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5341 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5401 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5461 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5521 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5581 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5641 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5701 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5761 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5821 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5881 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5941 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6001 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6061 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6121 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6181 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6241 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6301 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6361 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6421 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6481 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6541 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6601 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6661 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6721 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6781 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6841 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6901 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6961 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7021 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     7081 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     7141 tttatgcttc cgcggctcgt atgttgtgtg gaattgtgag cggataacaa tttcaca
//
