LOCUS       sequence-152117-        5717 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 5717)
  AUTHORS   Dower K, Rosbash M
  TITLE     T7 RNA polymerase-directed transcripts are processed in yeast and 
            link 3' end formation to mRNA nuclear export.
  JOURNAL   RNA. 2002 May;8(5):686-97.
  PUBMED    12022234
REFERENCE   2  (bases 1 to 5717)
  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: "RNA."; 
            date: "2002-05"; volume: "8(5)"; pages: "686-97"
COMMENT     Sequence Label: pRS314-T7-GFP
FEATURES             Location/Qualifiers
     source          1..5717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    1995..2498
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2539..2557
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2595..2617)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2717..2736
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2757..3037
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             3038..3712
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(3815..4270)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3952..3973)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4164..4183
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4396..4418
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4410..4427
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4411..4427
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4434..4452
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4485..4503
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4549..5265
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(4582..4610)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5205..5225
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      5398..5444
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5465..5481)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5466..5482)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(5510..5528)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5549..5565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5549..5565)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5562..5584)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5573..5589
                     /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(5597..5603,5604..5621,5622..5627))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5642..5663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgggtcct tttcatcacg tgctataaaa ataattataa tttaaatttt ttaatataaa
     2041 tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc
     2101 cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat cttgctcttc
     2161 ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga ccacacacga
     2221 aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat ttaatctgct
     2281 tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt
     2341 gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact ttctaaaatc
     2401 caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat tccatcatta
     2461 aaagatacga ggcgcgtgta agttacaggc aagcgatccg tctaagaaac cattattatc
     2521 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     2581 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     2641 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     2701 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tagatcacga
     2761 cattactata tatataatat aggaagcatt taatagaaca gcatcgtaat atatgtgtac
     2821 tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa aaatagcttg
     2881 tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa gaattaattc
     2941 ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat tgagcacgtg
     3001 agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta atttcacagg
     3061 tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg cagaatgtgc
     3121 tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa agagaacaat
     3181 tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa atagttcagg
     3241 cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg ttttggctct
     3301 ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt ggcaagaata
     3361 ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag actgcaacat
     3421 actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg attcagaagc
     3481 aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg gaaggcaaga
     3541 gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa atgttggtga
     3601 tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg
     3661 tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat aggttattac
     3721 tgagtagtat ttatttaagt attgtttgtg cacttgccga tctatgcggt gtgaaatacc
     3781 gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta
     3841 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3901 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3961 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4021 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4081 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4141 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4201 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4261 cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4321 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     4381 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg
     4441 actcactata gggcgaattg gagctccacc gcggtggcgg ccgctaatac gactcactat
     4501 agggagaact tagtttcgaa taaacacaca taaacaaaca aaggatccat ggctagcaaa
     4561 ggagaagaac tcttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat
     4621 gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc
     4681 cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact
     4741 ttcacttatg gtgttcagtg cttttcaaga tacccggatc atatgaaacg gcatgacttt
     4801 ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac
     4861 gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc
     4921 gagttaaaag gtattgattt taaagaagat ggaaacattc ttgggcacaa attggaatac
     4981 aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagct
     5041 aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa
     5101 caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca
     5161 caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt
     5221 gtaacagctg ctgggattac acatggcatg gatgaactat acaaaggatc tggctgctaa
     5281 gtcgactaag tgaatttact ttaaatcttg catttaaata aattttcttt ttatagcttt
     5341 atgacttagt ctcaatttat atactatttt aatgacattt tgattcattg attgaaatag
     5401 cataacccct tggggcctct aaacgggtct tgaggggttt tttggaattc gatatcaagc
     5461 ttatcgatac cgtcgacctc gagggggggc ccggtaccag cttttgttcc ctttagtgag
     5521 ggttaatttc gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5581 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5641 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5701 acctgtcgtg ccagctg
//
