LOCUS       sequence-366057-        4961 bp    DNA     circular SYN 14-APR-2025
DEFINITION  A pEvol-based plasmid containing one inducible (by arabinose) copy 
            of the Mut1-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 Apr 14, 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: Mut1-RS
FEATURES             Location/Qualifiers
     source          1..4961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(288..305)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(422..967)
                     /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(502..521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1113..1991)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2018..2302
                     /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     2225..2244
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3269..3286
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3342..3359)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(4095..4754)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4669..4688
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4755..4857)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt tagtgtatga tggtgttttt
       61 gaggtgctcc agtggcttct gtttctatca gctgtccctc ctgttcagct actgacgggg
      121 tggtgcgtaa cggcaaaagc accgccggac atcagcgcta gcggagtgta tactggctta
      181 ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct
      241 gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac
      301 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat
      361 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt
      421 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt
      481 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc
      541 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt
      601 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg
      661 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      721 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
      781 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
      841 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
      901 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
      961 atctcaagaa gatcatctta ttaatcagat aaaatatttc tagatttcag tgcaatttat
     1021 ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgttt
     1081 gacagcttat catcgataag cttggtaccc aattatgaca acttgacggc tacatcattc
     1141 actttttctt cacaaccggc acggaactcg ctcgggctgg ccccggtgca ttttttaaat
     1201 acccgcgaga aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc
     1261 atccgggtgg tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt
     1321 aagacgctaa tccctaactg ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa
     1381 acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac
     1441 tgacaagcct cgcgtacccg attatccatc ggtggatgga gcgactcgtt aatcgcttcc
     1501 atgcgccgca gtaacaattg ctcaagcaga tttatcgcca gcagctccga atagcgccct
     1561 tccccttgcc cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct
     1621 tcatccgggc gaaagaaccc cgtattggca aatattgacg gccagttaag ccattcatgc
     1681 cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc attcgcgagc ctccggatga
     1741 cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa tatcactcgg tcggcaaaca
     1801 aattctcgtc cctgattttt caccaccccc tgaccgcgaa tggtgagatt gagaatataa
     1861 cctttcattc ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc
     1921 gttaaacccg ccaccagatg ggcattaaac gagtatcccg gcagcagggg atcattttgc
     1981 gcttcagcca tacttttcat actcccgcca ttcagagaag aaaccaattg tccatattgc
     2041 atcagacatt gccgtcactg cgtcttttac tggctcttct cgctaaccaa accggtaacc
     2101 ccgcttatta aaagcattct gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac
     2161 aaaagtgtct ataatcacgg cagaaaagtc cacattgatt atttgcacgg cgtcacactt
     2221 tgctatgcca tagcattttt atccataaga ttagcggatc ctacctgacg ctttttatcg
     2281 caactctcta ctgtttctcc atacccgttt ttttgggcta acaggaggaa ttagatctat
     2341 ggacgaattt gaaatgataa agagaaacac atctgaaatt atcagcgagg aagagttaag
     2401 agaggtttta aaaaaagatg aaaaatctgc tctgataggt tttgaaccaa gtggtaaaat
     2461 acatttaggg cattatctcc aaataaaaaa gatgattgat ttacaaaatg ctggatttga
     2521 tataattata ttgttggctg atttacacgc ctatttaaac cagaaaggag agttggatga
     2581 gattagaaaa ataggagatt ataacaaaaa agtttttgaa gcaatggggt taaaggcaaa
     2641 atatgtttat ggaagtgaat tccagcttga taaggattat acactgaatg tctatagatt
     2701 ggctttaaaa actaccttaa aaagagcaag aaggagtatg gaacttatag caagagagga
     2761 tgaaaatcca aaggttgctg aagttatcta tccaataatg caggttaatt ctatgcatta
     2821 taagggcgtt gatgttcatg ttggagggat ggagcagaga aaaatacaca tgttagcaag
     2881 ggagctttta ccaaaaaagg ttgtttgtat tcacaaccct gtcttaacgg gtttggatgg
     2941 agaaggaaag atgagttctt caaaagggaa ttttatagct gttgatgact ctccagaaga
     3001 gattagggct aagataaaga aagcatactg cccagctgga gttgttgaag gaaatccaat
     3061 aatggagata gctaaatact tccttgaata tcctttaacc ataaaagggc cagaaaaatt
     3121 tggtggagat ttgacagtta atagctatga ggagttagag agtttattta aaaataagga
     3181 attgcatcca atgcgcttaa aaaatgctgt agctgaagaa cttataaaga ttttagagcc
     3241 aattagaaag agattataat aagtcgacca tcatcatcat catcattgag tttaaacggt
     3301 ctccagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     3361 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     3421 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     3481 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     3541 ctgggccttg tttgtgagct cccggtcatc aatcatcccc ataatccttg ttagattatc
     3601 aaattctgct tttcttcgcg aattaattcc gcttcgcaac atgtgagcac cggtttattg
     3661 actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt tgctcaggct
     3721 ctccccgtgg aggtaataat tgacgatatg atcagtgcac ggctaactaa gcggcctgct
     3781 gactttctcg ccgatcaaaa ggcattttgc tattaaggga ttgacgaggg cgtatctgcg
     3841 cagtaagatg cgccccgcat tccggcggta gttcagcagg gcagaacggc ggactctaaa
     3901 tccgcatggc aggggttcaa atcccctccg ccggaccaaa ttcgaaaagc ctgctcaacg
     3961 agcaggcttt tttgcatgct cgagcagctc agggtcgaat ttgctttcga atttctgcca
     4021 ttcatccgct tattatcact tattcaggcg tagcaaccag gcgtttaagg gcaccaataa
     4081 ctgccttaaa aaaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta
     4141 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
     4201 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
     4261 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
     4321 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
     4381 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
     4441 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
     4501 catatcacca gctcaccgtc tttcattgcc atacggaatt ccggatgagc attcatcagg
     4561 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
     4621 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
     4681 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
     4741 atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
     4801 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg
     4861 tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt
     4921 atttattctg cgaagtgatc ttccgtcaca ggtatttatt c
//
