LOCUS       sequence-62617-e        6644 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 6644)
  AUTHORS   Procko E, Hedman R, Hamilton K, Seetharaman J, Fleishman SJ, Su M, 
            Aramini J, Kornhaber G, Hunt JF, Tong L, Montelione GT, Baker D
  TITLE     Computational design of a protein-based enzyme inhibitor.
  JOURNAL   J Mol Biol. 2013 Sep 23;425(18):3563-75. doi: 
            10.1016/j.jmb.2013.06.035.
  PUBMED    23827138
REFERENCE   2  (bases 1 to 6644)
  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.jmb.2013.06"; journalName: "J Mol Biol"; date: 
            "2013-09-23- 23"; volume: "425"; issue: "18"; pages: "3563-75"
COMMENT     Sequence Label: pETCON-DnvLB16 M40R;W41H;H42S
FEATURES             Location/Qualifiers
     source          1..6644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /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     complement(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /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     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2940..3604
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    3156..3273
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3568..3591
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             3923..3934
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3935..3961
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4406..4435
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4764..4783)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4765..4783)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4790..4807)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4790..4806)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4799..4821)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4951..5406
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5038..5057)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5248..5269
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5503..6177)
                     /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"
     promoter        complement(6178..6458)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(6475..6494)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6594..6616
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt ttttcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 ggcagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc caggggggaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc cgagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctcggaatt
     2761 aaccctcact aaagggaaca aaagctgggt acccgacagg ttatcagcaa caacacagtc
     2821 atatccattc tcaattagct ctaccacagt gtgtgaacca atgtatccag caccacctgt
     2881 aaccaaaaca attttagaag tactttcact ttgtaactga gctgtcattt atattgaatt
     2941 ttcaaaaatt cttacttttt ttttggatgg acgcaaagaa gtttaataat catattacat
     3001 ggcattacca ccatatacat atccatatac atatccatat ctaatcttac ttatatgttg
     3061 tggaaatgta aagagcccca ttatcttagc ctaaaaaaac cttctctttg gaactttcag
     3121 taatacgctt aactgctcat tgctatattg aagtacggat tagaagccgc cgagcgggtg
     3181 acagccctcc gaaggaagac tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg
     3241 aaacgcagat gtgcctcgcg ccgcactgct ccgaacaata aagattctac aatactagct
     3301 tttatggtta tgaagaggaa aaattggcag taacctggcc ccacaaacct tcaaatgaac
     3361 gaatcaaatt aacaaccata ggatgataat gcgattagtt ttttagcctt atttctgggg
     3421 taattaatca gcgaagcgat gatttttgat ctattaacag atatataaat gcaaaaactg
     3481 cataaccact ttaactaata ctttcaacat tttcggtttg tattacttct tattcaaatg
     3541 taataaaagt atcaacaaaa aattgttaat atacctctat actttaacgt caaggagaaa
     3601 aaaccccgga tcgaattccc tacttcatac attttcaatt aagatgcagt tacttcgctg
     3661 tttttcaata ttttctgtta ttgcttcagt tttagcacag gaactgacaa ctatatgcga
     3721 gcaaatcccc tcaccaactt tagaatcgac gccgtactct ttgtcaacga ctactatttt
     3781 ggccaacggg aaggcaatgc aaggagtttt tgaatattac aaatcagtaa cgtttgtcag
     3841 taattgcggt tctcacccct caacaactag caaaggcagc cccataaaca cacagtatgt
     3901 ttttaaggac aatagctcga cgattgaagg tagataccca tacgacgttc cagactacgc
     3961 tctgcaggct agtggtggag gaggctctgg tggaggcggt agcggaggcg gagggtcggc
     4021 tagccatatg agattattag ccatcgaatg gagattgtgg gccttttaca tccctttgat
     4081 tgaagccttt tcctcttacg tttactacac cgccttagaa gatgaaatcg aattcattat
     4141 ccaaactagg cactctttgg ctggtaacaa cttgttgaga gttttgccaa tgcacttcaa
     4201 attcaaggct gacaaaattg ctggttctgc aaactggaca ttctacgcca aggaagattt
     4261 cgaacaatgg aaggaagctt tggacgtttt gtgggaagaa ttcttggaag tattgaagga
     4321 atattacaca ttgagtggta ctgaaatctc cttgcctgaa aagcctgact ggttgatagg
     4381 ttacaaactc gaggggggcg gatccgaaca aaagcttatt tctgaagagg acttgtaata
     4441 gagatctgat aacaacagtg tagatgtaac aaaatcgact ttgttcccac tgtactttta
     4501 gctcgtacaa aatacaatat acttttcatt tctccgtaaa caacatgttt tcccatgtaa
     4561 tatccttttc tatttttcgt tccgttacca actttacaca tactttatat agctattcac
     4621 ttctatacac taaaaaacta agacaatttt aattttgctg cctgccatat ttcaatttgt
     4681 tataaattcc tataatttat cctattagta gctaaaaaaa gatgaatgtg aatcgaatcc
     4741 taagagaatt gagctccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg
     4801 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4861 atcccccctt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4921 agttgcgcag cctgaatggc gaatggcgcg acgcgccctg tagcggcgca ttaagcgcgg
     4981 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5041 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5101 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5161 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5221 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5281 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5341 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     5401 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaggca
     5461 agtgcacaaa caatacttaa ataaatacta ctcagtaata acctatttct tagcattttt
     5521 gacgaaattt gctattttgt tagagtcttt tacaccattt gtctccacac ctccgcttac
     5581 atcaacacca ataacgccat ttaatctaag cgcatcacca acattttctg gcgtcagtcc
     5641 accagctaac ataaaatgta agctttcggg gctctcttgc cttccaaccc agtcagaaat
     5701 cgagttccaa tccaaaagtt cacctgtccc acctgcttct gaatcaaaca agggaataaa
     5761 cgaatgaggt ttctgtgaag ctgcactgag tagtatgttg cagtcttttg gaaatacgag
     5821 tcttttaata actggcaaac cgaggaactc ttggtattct tgccacgact catctccatg
     5881 cagttggacg atatcaatgc cgtaatcatt gaccagagcc aaaacatcct ccttaggttg
     5941 attacgaaac acgccaacca agtatttcgg agtgcctgaa ctatttttat atgcttttac
     6001 aagacttgaa attttccttg caataaccgg gtcaattgtt ctctttctat tgggcacaca
     6061 tataataccc agcaagtcag catcggaatc tagagcacat tctgcggcct ctgtgctctg
     6121 caagccgcaa actttcacca atggaccaga actacctgtg aaattaataa cagacatact
     6181 ccaagctgcc tttgtgtgct taatcacgta tactcacgtg ctcaatagtc accaatgccc
     6241 tccctcttgg ccctctcctt ttcttttttc gaccgaatta attcttaatc ggcaaaaaaa
     6301 gaaaagctcc ggatcaagat tgtacgtaag gtgacaagct atttttcaat aaagaatatc
     6361 ttccactact gccatctggc gtcataactg caaagtacac atatattacg atgctgtcta
     6421 ttaaatgctt cctatattat atatatagta atgtcgttta tggtgcactc tcagtacaat
     6481 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     6541 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     6601 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcga
//
