LOCUS       sequence-62614-e        6767 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 6767)
  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 6767)
  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-HtsptLB12v.3
FEATURES             Location/Qualifiers
     source          1..6767
                     /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             4529..4558
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4887..4906)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4888..4906)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4913..4930)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4913..4929)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4922..4944)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5074..5529
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5161..5180)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5371..5392
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5626..6300)
                     /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(6301..6581)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(6598..6617)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6717..6739
                     /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 ggttccgaaa tgagtgaaat gtctgttgta gattacgtcg ttaagtcttt
     4081 taattataca gaggatatga gtcatgaatt gttattctct gttaaaaaga gatggttcgt
     4141 aaagccattc gaacactcaa ccagatatgg tttgttagcc tacatgttgt tacctggtaa
     4201 ctacataaag ttcggtttgt acgttttaac gaaccaagac tatgctagat tcgaaatagc
     4261 atgggtacat gtcgataaag acggtaaaat cgaagaaaga accgtatatt ccattgaaac
     4321 ttactggcac attttcattg atatcgaaaa cgacttgaac tgtccatacg ttttggcaaa
     4381 gtttattgaa atgagacctg aattcgataa gacagcctgg gtcgaagaat caaactattc
     4441 cattgctgaa gatgacatcc aaatggttga aagtataaaa agatacttgg aaagaaagat
     4501 cgcttctgat ctcgaggggg gcggatccga acaaaagctt atttctgaag aggacttgta
     4561 atagagatct gataacaaca gtgtagatgt aacaaaatcg actttgttcc cactgtactt
     4621 ttagctcgta caaaatacaa tatacttttc atttctccgt aaacaacatg ttttcccatg
     4681 taatatcctt ttctattttt cgttccgtta ccaactttac acatacttta tatagctatt
     4741 cacttctata cactaaaaaa ctaagacaat tttaattttg ctgcctgcca tatttcaatt
     4801 tgttataaat tcctataatt tatcctatta gtagctaaaa aaagatgaat gtgaatcgaa
     4861 tcctaagaga attgagctcc aattcgccct atagtgagtc gtattacaat tcactggccg
     4921 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     4981 cacatccccc cttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     5041 aacagttgcg cagcctgaat ggcgaatggc gcgacgcgcc ctgtagcggc gcattaagcg
     5101 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     5161 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     5221 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5281 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     5341 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5401 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     5461 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     5521 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcag
     5581 gcaagtgcac aaacaatact taaataaata ctactcagta ataacctatt tcttagcatt
     5641 tttgacgaaa tttgctattt tgttagagtc ttttacacca tttgtctcca cacctccgct
     5701 tacatcaaca ccaataacgc catttaatct aagcgcatca ccaacatttt ctggcgtcag
     5761 tccaccagct aacataaaat gtaagctttc ggggctctct tgccttccaa cccagtcaga
     5821 aatcgagttc caatccaaaa gttcacctgt cccacctgct tctgaatcaa acaagggaat
     5881 aaacgaatga ggtttctgtg aagctgcact gagtagtatg ttgcagtctt ttggaaatac
     5941 gagtctttta ataactggca aaccgaggaa ctcttggtat tcttgccacg actcatctcc
     6001 atgcagttgg acgatatcaa tgccgtaatc attgaccaga gccaaaacat cctccttagg
     6061 ttgattacga aacacgccaa ccaagtattt cggagtgcct gaactatttt tatatgcttt
     6121 tacaagactt gaaattttcc ttgcaataac cgggtcaatt gttctctttc tattgggcac
     6181 acatataata cccagcaagt cagcatcgga atctagagca cattctgcgg cctctgtgct
     6241 ctgcaagccg caaactttca ccaatggacc agaactacct gtgaaattaa taacagacat
     6301 actccaagct gcctttgtgt gcttaatcac gtatactcac gtgctcaata gtcaccaatg
     6361 ccctccctct tggccctctc cttttctttt ttcgaccgaa ttaattctta atcggcaaaa
     6421 aaagaaaagc tccggatcaa gattgtacgt aaggtgacaa gctatttttc aataaagaat
     6481 atcttccact actgccatct ggcgtcataa ctgcaaagta cacatatatt acgatgctgt
     6541 ctattaaatg cttcctatat tatatatata gtaatgtcgt ttatggtgca ctctcagtac
     6601 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     6661 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     6721 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
