LOCUS       sequence-366061-        4961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A pEvol-based plasmid containing one inducible (by arabinose) copy 
            of the Mut2-RS aaRS. The 2nd constitutive aaRS copy typically found 
            in pEvol plasmids was deleted..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4961)
  AUTHORS   Hadar D, Gelkop S, Vaserman L, Amiram M
  TITLE     Efficient Incorporation of Clickable Unnatural Amino Acids Enables 
            Rapid and Biocompatible Labeling of Proteins in Vitro and in 
            Bacteria.
  JOURNAL   Chembiochem. 2021 Apr 16;22(8):1379-1384. doi: 
            10.1002/cbic.202000663. Epub 2021 Feb 4.
  PUBMED    33350556
REFERENCE   2  (bases 1 to 4961)
  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/cbic.202000663"; journalName: "Chembiochem"; date: 
            "2021-04-16- 16"; volume: "22"; issue: "8"; pages: "1379-1384"
COMMENT     Sequence Label: Mut2-RS
FEATURES             Location/Qualifiers
     source          1..4961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(571..1449)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1476..1760
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1683..1702
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2727..2744
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2800..2817)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(3553..4212)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4127..4146
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4213..4315)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4707..4724)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4841..4961,1..425))
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4921..4940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct
       61 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
      121 cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      181 cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt
      241 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
      301 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac
      361 cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
      421 ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct
      481 cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgtttga
      541 cagcttatca tcgataagct tggtacccaa ttatgacaac ttgacggcta catcattcac
      601 tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac
      661 ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat
      721 ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa
      781 gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac
      841 atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg
      901 acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat
      961 gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc
     1021 cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc
     1081 atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca
     1141 gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg
     1201 accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcactcggtc ggcaaacaaa
     1261 ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc
     1321 tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt
     1381 taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc
     1441 ttcagccata cttttcatac tcccgccatt cagagaagaa accaattgtc catattgcat
     1501 cagacattgc cgtcactgcg tcttttactg gctcttctcg ctaaccaaac cggtaacccc
     1561 gcttattaaa agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa
     1621 aagtgtctat aatcacggca gaaaagtcca cattgattat ttgcacggcg tcacactttg
     1681 ctatgccata gcatttttat ccataagatt agcggatcct acctgacgct ttttatcgca
     1741 actctctact gtttctccat acccgttttt ttgggctaac aggaggaatt agatctatgg
     1801 acgaatttga aatgataaag agaaacacat ctgaaattat cagcgaggaa gagttaagag
     1861 aggttttaaa aaaagatgaa aaatctgctc tgataggttt tgaaccaagt ggtaaaatac
     1921 atttagggca ttatctccaa ataaaaaaga tgattgattt acaaaatgct ggatttgata
     1981 taattatagt tttggctgat ttacacgcct atttaaacca gaaaggagag ttggatgaga
     2041 ttagaaaaat aggagattat aacaaaaaag tttttgaagc aatggggtta aaggcaaaat
     2101 atgtttatgg aagtgaattc cagcttgata aggattatac actgaatgtc tatagattgg
     2161 ctttaaaaac taccttaaaa agagcaagaa ggagtatgga acttatagca agagaggatg
     2221 aaaatccaaa ggttgctgaa gttatctatc caataatgca ggttaatggg gctcattatg
     2281 agggcgttga tgttcatgtt ggagggatgg agcagagaaa aatacacatg ttagcaaggg
     2341 agcttttacc aaaaaaggtt gtttgtattc acaaccctgt cttaacgggt ttggatggag
     2401 aaggaaagat gagttcttca aaagggaatt ttatagctgt tgatgactct ccagaagaga
     2461 ttagggctaa gataaagaaa gcatactgcc cagctggagt tgttgaagga aatccaataa
     2521 tggagatagc taaatacttc cttgaatatc ctttaaccat aaaagggcca gaaaaatttg
     2581 gtggagattt gacagttaat agctatgagg agttagagag tttatttaaa aataaggaat
     2641 tgcatccaat gcgcttaaaa aatgctgtag ctgaagaact tataaagatt ttagagccaa
     2701 ttagaaagag attataataa gtcgaccatc atcatcatca tcattgagtt taaacggtct
     2761 ccagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
     2821 acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
     2881 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
     2941 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     3001 gggccttgtt tgtgagctcc cggtcatcaa tcatccccat aatccttgtt agattatcaa
     3061 attctgcttt tcttcgcgaa ttaattccgc ttcgcaacat gtgagcaccg gtttattgac
     3121 taccggaagc agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg ctcaggctct
     3181 ccccgtggag gtaataattg acgatatgat cagtgcacgg ctaactaagc ggcctgctga
     3241 ctttctcgcc gatcaaaagg cattttgcta ttaagggatt gacgagggcg tatctgcgca
     3301 gtaagatgcg ccccgcattc cggcggtagt tcagcagggc agaacggcgg actctaaatc
     3361 cgcatggcag gggttcaaat cccctccgcc ggaccaaatt cgaaaagcct gctcaacgag
     3421 caggcttttt tgcatgctcg agcagctcag ggtcgaattt gctttcgaat ttctgccatt
     3481 catccgctta ttatcactta ttcaggcgta gcaaccaggc gtttaagggc accaataact
     3541 gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag
     3601 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
     3661 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt
     3721 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac
     3781 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc
     3841 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     3901 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
     3961 tatcaccagc tcaccgtctt tcattgccat acggaattcc ggatgagcat tcatcaggcg
     4021 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
     4081 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa
     4141 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
     4201 ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     4261 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc
     4321 tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac caggatttat
     4381 ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga
     4441 tgctgccaac ttactgattt agtgtatgat ggtgtttttg aggtgctcca gtggcttctg
     4501 tttctatcag ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac ggcaaaagca
     4561 ccgccggaca tcagcgctag cggagtgtat actggcttac tatgttggca ctgatgaggg
     4621 tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat
     4681 atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
     4741 ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag
     4801 atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc
     4861 ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat
     4921 aaagatacca ggcgtttccc cctggcggct ccctcgtgcg c
//
