LOCUS       sequence-52600-e        6507 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 6507)
  AUTHORS   Chu D, Kazana E, Bellanger N, Singh T, Tuite MF, von der Haar T
  TITLE     Translation elongation can control translation initiation on 
            eukaryotic mRNAs.
  JOURNAL   EMBO J. 2014 Jan 1;33(1):21-34. doi: 10.1002/embj.201385651. Epub 
            2013 Dec 19.
  PUBMED    24357599
REFERENCE   2  (bases 1 to 6507)
  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: 
            "10.1002/embj.201385651"; journalName: "EMBO J"; date: "2014-01-1- 
            1"; volume: "33"; issue: "1"; pages: "21-34"
COMMENT     Sequence Label: pTH761-CEN-mCherry_v4
FEATURES             Location/Qualifiers
     source          1..6507
                     /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"
     promoter        190..410
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             411..1214
                     /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(1343..1798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1480..1501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1692..1711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1928..1950
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1942..1959
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1943..1959
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2005..2648
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2542..2565
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     terminator      3383..3630
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3399..3417)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     3637..3653
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3637..3653
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3687..3703)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3688..3704)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(3733..3751)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3772..3788)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3772..3788)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3785..3807)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3796..3812
                     /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(3820..3826,3827..3844,3845..3850))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4003..4020)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4174..4762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4933..5724,5725..5793))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5556..5575
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5794..5898)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5935..6438
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6480..6498
                     /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 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
      241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
      301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
      361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
      421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
      481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
      541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
      601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
      661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
      721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
      781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
      841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
      901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
      961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     1021 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     1081 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     1261 gagcttcaat ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt
     1321 aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta
     1381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     1441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     1501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     1561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     1621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     1681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     1741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg
     1801 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1861 ctattacgcc agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca
     1921 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     1981 tagggcgaat tggagctcag tttatcatta tcaatactcg ccatttcaaa gaatacgtaa
     2041 ataattaata gtagtgattt tcctaacttt atttagtcaa aaaattagcc ttttaattct
     2101 gctgtaaccc gtacatgccc aaaatagggg gcgggttaca cagaatatat aacatcgtag
     2161 gtgtctgggt gaacagttta ttcctggcat ccactaaata taatggagcc cgctttttaa
     2221 gctggcatcc agaaaaaaaa agaatcccag caccaaaata ttgttttctt caccaaccat
     2281 cagttcatag gtccattctc ttagcgcaac tacagagaac aggggcacaa acaggcaaaa
     2341 aacgggcaca acctcaatgg agtgatgcaa cctgcctgga gtaaatgatg acacaaggca
     2401 attgacccac gcatgtatct atctcatttt cttacacctt ctattacctt ctgctctctc
     2461 tgatttggaa aaagctgaaa aaaaaggttg aaaccagttc cctgaaatta ttcccctact
     2521 tgactaataa gtatataaag acggtaggta ttgattgtaa ttctgtaaat ctatttctta
     2581 aacttcttaa attctacttt tatagttagt ctttttttta gttttaaaac accagaactt
     2641 agtttcgacg gattctagaa ctagtatggt ttcaaagggc gaagaagaca atatggctat
     2701 tattaaggaa ttcatgagat tcaaggtcca catggaaggt tctgtcaacg gtcacgaatt
     2761 cgaaattgaa ggtgaaggtg aaggtagacc atacgaaggt acccaaaccg ctaagttgaa
     2821 ggtcaccaag ggtggtccat tgccattcgc ttgggatatt ttgtctccac aattcatgta
     2881 cggttctaag gcttacgtca agcacccagc tgatattcca gattacttga agttgtcttt
     2941 cccagaaggt ttcaagtggg aaagagtcat gaacttcgaa gatggtggtg tcgtcaccgt
     3001 cacccaagat tcttctttgc aagatggtga attcatttac aaggtcaagt tgagaggtac
     3061 caacttccca tctgatggtc cagtcatgca aaagaagacc atgggttggg aagcttcttc
     3121 tgaaagaatg tacccagaag atggtgcttt gaagggtgaa attaagcaaa gattgaagtt
     3181 gaaggatggt ggtcactacg atgctgaagt caagaccacc tacaaggcta agaagccagt
     3241 ccaattgcca ggtgcttaca acgtcaacat taagttggat attacctctc acaacgaaga
     3301 ttacaccatt gtcgaacaat acgaaagagc tgaaggtaga cactctaccg gtggtatgga
     3361 tgaattgtac aagtaactcg agtcatgtaa ttagttatgt cacgcttaca ttcacgccct
     3421 ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     3481 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     3541 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
     3601 gttttgggac gctcgaaggc tttaatttgc ggccgctcta gaactagtgg atcccccggg
     3661 ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc tcgagggggg gcccggtacc
     3721 cagcttttgt tccctttagt gagggttaat tccgagcttg gcgtaatcat ggtcatagct
     3781 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacataggag ccggaagcat
     3841 aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc acattaattg cgttgcgctc
     3901 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3961 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4021 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4081 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4141 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctcggcc cccctgacga
     4201 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4261 ccaggcgttc ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4321 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     4381 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4441 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4501 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4561 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     4621 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4681 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4741 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4801 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4861 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4921 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4981 tcgttcatcc atagttgcct gactgcccgt cgtgtagata actacgatac gggagggctt
     5041 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5101 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5161 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5221 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5281 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5341 gtgaaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5401 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5461 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5521 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5581 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5641 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5701 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5761 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5821 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5881 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcct tttcatcacg
     5941 tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt aaaaatagaa
     6001 agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta aaagactcta
     6061 gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag gtattaatgc
     6121 cgaattgttt catcttgtct gtgtagaaga ccacacacga aaatcctgtg attttacatt
     6181 ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct aataaatata
     6241 tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt ttttcttcat
     6301 tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa acataaaaat
     6361 aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga ggcgcgtgta
     6421 agttacaggc aagcgatccg tcctaagaaa ccattattat catgacatta acctataaaa
     6481 ataggcgtat cacgaggccc tttcgtc
//
