LOCUS       sequence-218749-        7582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MET25p-MS2-RFP-CYC1 TT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7582)
  AUTHORS   Clemens K, Bilanchone V, Beliakova-Bethell N, Larsen LS, Nguyen K, 
            Sandmeyer S
  TITLE     Sequence requirements for localization and packaging of Ty3 
            retroelement RNA.
  JOURNAL   Virus Res. 2013 Feb;171(2):319-31. doi: 
            10.1016/j.virusres.2012.10.008. Epub 2012 Oct 13.
  PUBMED    23073180
REFERENCE   2  (bases 1 to 7582)
  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.virusres.2012.10.008"; journalName: "Virus Res"; date: 
            "2013-02"; volume: "171"; issue: "2"; pages: "319-31"
COMMENT     Sequence Label: pNB2629
FEATURES             Location/Qualifiers
     source          1..7582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(664..1758)
                     /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"
     promoter        complement(1759..2166)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2467..2922)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2604..2625)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2816..2835
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3048..3070
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3062..3079
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3063..3079
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3086..3105
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3086..3104
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3151..3500
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     complement(4573..4591)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      4603..4792
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(4808..4826)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4847..4863)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4847..4863)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4860..4882)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4871..4887
                     /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)."
     promoter        complement(join(4895..4901,4902..4919,4920..4925))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4940..4961
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5078..5095)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5249..5837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5329..5348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6008..6799,6800..6868))
                     /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     6631..6650
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6869..6973)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7010..7513
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7555..7573
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatcga ctacgtcgtt aaggccgttt ctgacagagt aaaattcttg agggaacttt
      241 caccattatg ggaaatggtt caagaaggta ttgacttaaa ctccatcaaa tggtcaggtc
      301 attgagtgtt ttttatttgt tgtatttttt tttttttaga gaaaatcctc caatatataa
      361 attaggaatc atagtttcat gattttctgt tacacctaac tttttgtgtg gtgccctcct
      421 ccttgtcaat attaatgtta aagtgcaatt ctttttcctt atcacgttga gccattagta
      481 tcaatttgct tacctgtatt cctttacatc ctcctttttc tccttcttga taaatgtatg
      541 tagattgcgt atatagtttc gtctacccta tgaacatatt ccattttgta atttcgtgtc
      601 gtttctatta tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc
      661 tttttaagca aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact
      721 gttggaacca cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc
      781 ttcaatggcc ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga
      841 caagatagtg gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca
      901 tggttcgtac aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg
      961 caacaaaccc aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat
     1021 gttgctggtg attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc
     1081 agaatcaatc aattgatgtt gaaccttcaa tgtagggaat tcgttcttga tggtttcctc
     1141 cacagttttt ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat
     1201 aggcaatggt ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac
     1261 ttctggaacg gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct
     1321 cttaccaaag taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc
     1381 aaattgtggc ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct
     1441 taagttggcg tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac
     1501 actaccggta ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt
     1561 ggaggcttcc agcgcctcat ctggaagtgg aacacctgta gcatcgatag cagcaccacc
     1621 aattaaatga ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag
     1681 aaccttaatg gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat
     1741 cttcttaggg gcagacatta gaatggtata tccttgaaat atatatatat atattgctga
     1801 aatgtaaaag gtaagaaaag ttagaaagta agacgattgc taaccaccta ttggaaaaaa
     1861 caataggtcc ttaaataata ttgtcaactt caagtattgt gatgcaagca tttagtcatg
     1921 aacgcttctc tattctatat gaaaagccgg ttccggcgct ctcacctttc ctttttctcc
     1981 caatttttca gttgaaaaag gtatatgcgt caggcgacct ctgaaattaa caaaaaattt
     2041 ccagtcatcg aatttgattc tgtgcgatag cgcccctgtg tgttctcgtt atgttgagga
     2101 aaaaaataat ggttgctaag agattcgaac tcttgcatct tacgatacct gagtattccc
     2161 acagttaact gcggtcaaga tatttcttga atcaggcgcc ttagaccgct cggccaaaca
     2221 accaattact tgttgagaaa tagagtataa ttatcctata aatataacgt ttttgaacac
     2281 acatgaacaa ggaagtacag gacaattgat tttgaagaga atgtggattt tgatgtaatt
     2341 gttgggattc catttttaat aaggcaataa tattaggtat gtagatatac tagaagttct
     2401 cctcgaccgg tcgatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     2461 atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2521 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2581 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2641 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2701 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2761 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2821 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2881 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct
     2941 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     3001 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     3061 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctccg
     3121 gatgcaaggg ttcgaatccc ttagctctca ttattttttg ctttttctct tgaggtcaca
     3181 tgatcgcaaa atggcaaatg gcacgtgaag ctgtcgatat tggggaactg tggtggttgg
     3241 caaatgacta attaagttag tcaaggcgcc atcctcatga aaactgtgta acataataac
     3301 cgaagtgtcg aaaaggtggc accttgtcca attgaacacg ctcgatgaaa aaaataagat
     3361 atatataagg ttaagtaaag cgtctgttag aaaggaagtt tttccttttt cttgctctct
     3421 tgtcttttca tctactattt ccttcgtgta atacagggtc gtcagataca tagatacaat
     3481 tctattaccc ccatccatac tctagaatgg cttctaactt tactcagttc gttctcgtcg
     3541 acaatggcgg aactggcgac gtgactgtcg ccccaagcaa cttcgctaac gggatcgctg
     3601 aatggatcag ctctaactcg cgttcacagg cttacaaagt aacctgtagc gttcgtcaga
     3661 gctctgcgca gaatcgcaaa tacaccatca aagtcgaggt gcctaaaggc gcctggcgtt
     3721 cgtacttaaa tatggaacta accattccaa ttttcgccac gaattccgac tgcgagctta
     3781 ttgttaaggc aatgcaaggt ctcctaaaag atggaaaccc gattccctca gcaatcgcag
     3841 caaactccgg catctacgga tccatggaca acactgaaga cgttatcaag gaattcatgc
     3901 aattcaaggt tagaatggaa ggttctgtta acggtcacta cttcgaaatc gagggcgaag
     3961 gtgaaggtaa gccatacgaa ggtactcaaa ctgctaagtt gcaagttact aagggtggtc
     4021 cattgccatt cgcttgggac atcttgtctc cacaattcca atacggttct aaggcttacg
     4081 ttaaacatcc tgctgacatc ccagactaca tgaagttgtc tttcccagaa ggtttcacat
     4141 gggaacgttc tatgaacttc gaagacggtg gtgtggttga agttcaacaa gattcttctt
     4201 tgcaagacgg tactttcatc tacaaggtta agttcaaggg tgttaacttc ccagctgacg
     4261 gtccagttat gcaaaagaag actgctggtt gggaaccgtc taccgaaaag ttgtacccac
     4321 aagatggcgt tttgaaaggt gaaatctctc acgctttgaa gttgaaggac ggtgggcact
     4381 atacttgtga cttcaagact gtttacaaag ctaagaagcc agttcaattg ccaggtaacc
     4441 attacgttga ctctaagttg gacatcacta accacaacga agactacact gttgttgaac
     4501 aatacgaaca cgctgaagct agacactctg gttctcaata gctcgagctg gtcgagtcat
     4561 gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa
     4621 ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt
     4681 attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc
     4741 atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgggccc ggtaccagct
     4801 tttgttccct ttagtgaggg ttaatttcga gcttggcgta atcatggtca tagctgtttc
     4861 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4921 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4981 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5041 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5101 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5161 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5221 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5281 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5341 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5401 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5461 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5521 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5581 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5641 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5701 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5761 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5821 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5881 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5941 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6001 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6061 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6121 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6181 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6241 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6301 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6361 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6421 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6481 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6541 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6601 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6661 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6721 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6781 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6841 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6901 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6961 taggggttcc gcgcacattt ccccgaaaag tgccacctgg gtccttttca tcacgtgcta
     7021 taaaaataat tataatttaa attttttaat ataaatatat aaattaaaaa tagaaagtaa
     7081 aaaaagaaat taaagaaaaa atagtttttg ttttccgaag atgtaaaaga ctctaggggg
     7141 atcgccaaca aatactacct tttatcttgc tcttcctgct ctcaggtatt aatgccgaat
     7201 tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc ctgtgatttt acattttact
     7261 tatcgttaat cgaatgtata tctatttaat ctgcttttct tgtctaataa atatatatgt
     7321 aaagtacgct ttttgttgaa attttttaaa cctttgttta tttttttttc ttcattccgt
     7381 aactcttcta ccttctttat ttactttcta aaatccaaat acaaaacata aaaataaata
     7441 aacacagagt aaattcccaa attattccat cattaaaaga tacgaggcgc gtgtaagtta
     7501 caggcaagcg atccgtccta agaaaccatt attatcatga cattaaccta taaaaatagg
     7561 cgtatcacga ggccctttcg tc
//
