LOCUS       sequence-226824-        7549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7549)
  AUTHORS   Henry MF, Silver PA
  TITLE     A novel methyltransferase (Hmt1p) modifies poly(A)+-RNA-binding 
            proteins.
  JOURNAL   Mol Cell Biol. 1996 Jul . 16(7):3668-78.
  PUBMED    8668183
REFERENCE   2  (bases 1 to 7549)
  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"; journalName: "Mol Cell 
            Biol. 1996 Jul . 16(7):3668-78."
COMMENT     Sequence Label: pPS1307
FEATURES             Location/Qualifiers
     source          1..7549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2742..2758
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2742..2758
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2792..2808)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2793..2809)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2837..2855)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2876..2892)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2876..2892)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2889..2911)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2900..2916
                     /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(2924..2930,2931..2948,2949..2954))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2969..2990
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3107..3124)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3278..3866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3358..3377)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4037..4828,4829..4897))
                     /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     4660..4679
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4898..5002)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5039..5542
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5583..5601
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5639..5661)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5761..5780
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5804..6019
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             6020..6823
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(6954..7409)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7091..7112)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7303..7322
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(7535..7549,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7549,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg gagctccacc
       61 gcggtggcgg ccgctctaga aaggctcatg atattgctac caacccgaat gcggcaatcg
      121 tattcttttg gaaggatctg caaaggcagg tgagagttga aggtatcaca gagcatgtta
      181 acagagaaac ttctgaaaga tactttaaga cgagacctcg tggatccaag atcggtgcat
      241 gggcttcccg ccaatcggat gttatcaaga acagagaaga actagacgag ttgacccaaa
      301 aaaacaccga acgtttcaag gatgctgaag acatcccatg tccagattat tggggtggct
      361 tgagaatcgt tccactggaa attgagttct ggcaaggtag accctcgaga ttgcatgata
      421 gattcgttta cagaagaaaa acagaaaacg atccatggaa agtcgttaga ctagccccat
      481 gagggactgt taatgaaaaa ttcaatgtag gatctattta acaccatata attctctcat
      541 tttctatttt atactgtata atgaatcttt ttataccgcc tatagggtaa tatgtcagaa
      601 tataccgaga tgaaaaaaaa gaaaaaggaa ttccgaaatt tggaaatggc gggaaaattt
      661 tttcacaaag ggaaattata tgttataaaa atagttgaag agatgagctg cctactgctc
      721 accttgccgt ttccaaaaaa gagttagaac cgacaaattc atccaaagaa aataatgagc
      781 aagacagccg tgaaagattc tgctacagaa aaaaccaagc taagtgaaag cgaacagcac
      841 tacttcaatt cgtacgatca ctatggtatt cacgaagaga tgcttcaaga tactgttcgt
      901 accttatctt acagaaacgc aattatccaa aataaggatc tttttaagga caagattgtt
      961 ttagacgtcg gttgcggtac cggtatttta tccatgtttg ccgctaaaca cggtgcgaag
     1021 catgttatcg gtgttgatat gtcaagcatt attgagatgg cgaaggaatt ggtagagttg
     1081 aacggattca gcgacaagat caccttgcta agaggcaagt tggaggacgt tcatttaccc
     1141 tttcctaaag ttgacatcat aatttctgaa tggatgggtt actttctact atacgagtcc
     1201 atgatggaca ccgttcttta cgctagagac cactatttgg tagaaggcgg tctgatcttt
     1261 cccgacaagt gctccattca tttggccggt ttggaagact ctcagtataa agacgagaag
     1321 ttgaactact ggcaagacgt ttacgggttt gattattcgc catttgttcc gttggtctta
     1381 cacgagccca tcgtcgacac cgtggaaaga aacaatgtca acaccacctc agacaaattg
     1441 atcgaatttg atttaaatac agtaaaaata tcagatctag cgtttaagag taactttaaa
     1501 ttgacggcca agagacaaga tatgattaat ggtatagtca cctggttcga cattgttttc
     1561 cctgcaccaa agggtaagag acctgttgag ttctccactg gtcctcatgc tccatacact
     1621 cactggaagc aaacaatatt ttatttccct gatgatctag atgctgaaac tggtgacacc
     1681 attgaaggtg aattggtttg ctctccaaac gagaagaata acagagatct aaatatcaaa
     1741 atttcttaca agttcgaatc gaatggcatc gacggtaatt caagaagcag aaaaaacgaa
     1801 ggttcttatt taatgcatta agacttttgt ttgctggaga aagaaaaaaa tttgaaaagc
     1861 aaataaacaa acttgttcat attattgacg cgaatatgtg tactttgtta cccttttttg
     1921 ttggtcgtgt accaaaatta gtttagatat atgactttgc atgtccttgt actatagttt
     1981 ttaaaagaaa aaatctactt atctatcctt cattaggatg ctatttttca ccttgttatc
     2041 atgaatatta gttaaagaag caacagcaac aacccaattc tgaatcatcc atcaatttcc
     2101 aacctatgtc ggcaaacttt ttcaatatct ttttgttttc ttcctccgat ggcgcaacgt
     2161 ttgaggcgtt actagctgca cattttccgg aatacatgct cttcattacc gattcgtagc
     2221 atccggtact acttgtagca cttacatagt aacgcttagc acggtcatgt gcattcttcc
     2281 taaaaggttc tagggtatta aatagtgcgg ttatttgcac caattcgttc agcgcagttg
     2341 acaacgtcat aattctgtca tttattccct tggccatatt tccattacgg atgtataaac
     2401 ttattaatgt ggtaggtatg ggtcctatca aactaatgat attatagcat tcttccacca
     2461 ttggcagcat gaaaatagaa acagttccca tcaagtcttt ggttgtgcaa cgtgtatttt
     2521 cttcactggc aagatttgac gtaatacttt gtgttggcga cattaattct tcttcactgt
     2581 cctgagcaaa ataggcgttc ggaaggggct ttgaatccaa atcaacagag ttttctgcgt
     2641 tcatattagt aattgaatct aatatgcctg ataccagagt gatcattgag cagactagct
     2701 ctccgatatt tttcatttca gcatccgaat tgcagcaatt ttctagaact agtggatccc
     2761 ccgggctgca ggaattcgat atcaagctta tcgataccgt cgacctcgag ggggggcccg
     2821 gtaccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat
     2881 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2941 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3001 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3061 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3121 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3181 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3241 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3301 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3361 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3421 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3481 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3541 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3601 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3661 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3721 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3781 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3841 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3901 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3961 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4021 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4081 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4141 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4201 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4261 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4321 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4381 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4441 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4501 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4561 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4621 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4681 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4741 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4801 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4861 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4921 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4981 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg tccttttcat
     5041 cacgtgctat aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat
     5101 agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac
     5161 tctaggggga tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta
     5221 atgccgaatt gtttcatctt gtctgtgtag aagaccacac acgaaaatcc tgtgatttta
     5281 cattttactt atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa
     5341 tatatatgta aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct
     5401 tcattccgta actcttctac cttctttatt tactttctaa aatccaaata caaaacataa
     5461 aaataaataa acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg
     5521 tgtaagttac aggcaagcga tccgtctaag aaaccattat tatcatgaca ttaacctata
     5581 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     5641 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     5701 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     5761 cggcatcaga gcagattgta ctgagagtgc accataccac cttttcaatt catcattttt
     5821 tttttattct tttttttgat ttcggtttcc ttgaaatttt tttgattcgg taatctccga
     5881 acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag
     5941 tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacctg
     6001 caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc
     6061 tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc
     6121 ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa
     6181 aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt
     6241 taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgaag acagaaaatt
     6301 tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga
     6361 atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa
     6421 gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc
     6481 atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag
     6541 cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg
     6601 ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag acgcattggg
     6661 tcaacagtat agaaccgtgg atgatgtggt ctctacagga tctgacatta ttattgttgg
     6721 aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc
     6781 aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt
     6841 aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc agttattacc
     6901 ctatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     6961 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     7021 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     7081 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     7141 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     7201 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     7261 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     7321 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     7381 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     7441 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     7501 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
