LOCUS       sequence-57491-e        3194 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 3194)
  AUTHORS   Moroz OV, Moroz YS, Wu Y, Olsen AB, Cheng H, Mack KL, McLaughlin JM,
            Raymond EA, Zhezherya K, Roder H, Korendovych IV
  TITLE     A Single Mutation in a Regulatory Protein Produces Evolvable 
            Allosterically Regulated Catalyst of Nonnatural Reaction.
  JOURNAL   Angew Chem Int Ed Engl. 2013 Apr 29. doi: 10.1002/anie.201302339.
  PUBMED    23630096
REFERENCE   2  (bases 1 to 3194)
  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: "Angew Chem 
            Int Ed Engl. 2013 Apr 29. doi: 10.1002/anie.201302339."
COMMENT     Sequence Label: pEXP5-NT-5th R
FEATURES             Location/Qualifiers
     source          1..3194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             122..142
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(650..668)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      664..711
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(756..774)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        842..946
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(947..1015,1016..1807)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1165..1184)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1978..2566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2467..2486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2720..2737
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2817..2836)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacca tgtctggttc tcatcatcat catcatcatg gtagcagcgg
      121 cgaaaacctg tattttcagt cccttatggc tgatcaactg acagaagagc agattgcaga
      181 attcaaagaa gctttttcac tatttgacaa ggatggtgat ggtactataa ctacaaagga
      241 gttggggact gtgatgagat cacttggtca gaaccccaca gaagcagaat tacaggacat
      301 gatcaatgaa gtagacgctg atggcaatgg cacaattgac ttcccagagt ttctgacaat
      361 gatggcaaga aaaatgaaag atacagatag cgaagaagaa cttagagaag cgttccgtgt
      421 ggaagacaag gatggtaatg gttacattag tgctgcagaa cttcgtattg tgatgacaaa
      481 tcgtggggag aagctaacag atgaagaagt tgatgaaatg attagggaaa cagacattga
      541 tggtgatggt caagtaaact atgaagagtt tgtacaacgt atgacagcga agtgaagggt
      601 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
      661 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
      721 ggaactatat ccggataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
      781 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
      841 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      901 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      961 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1021 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1081 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1141 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     1201 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1261 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1321 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1381 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1441 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1501 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1561 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1621 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1681 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1741 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1801 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1861 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1921 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1981 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2041 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2101 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2161 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2221 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2281 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2341 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2401 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2461 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2521 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2581 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2641 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2701 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2761 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca
     2821 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgcttcgtta
     2881 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     2941 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     3001 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     3061 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     3121 tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatct
     3181 cgatcccgcg aaat
//
