LOCUS       sequence-154225-        4538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSALECT plasmid with levoglucosan kinase fused to TEM-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4538)
  AUTHORS   Klesmith JR, Bacik JP, Wrenbeck EE, Michalczyk R, Whitehead TA
  TITLE     Trade-offs between enzyme fitness and solubility illuminated by deep
            mutational scanning.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Feb 14. pii: 201614437. doi: 
            10.1073/pnas.1614437114.
  PUBMED    28196882
REFERENCE   2  (bases 1 to 4538)
  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: "Proc Natl 
            Acad Sci U S A. 2017 Feb 14. pii: 201614437. doi: 
            10.1073/pnas.1614437114."
COMMENT     Sequence Label: pSALECTNK-LGK/csTEM1
FEATURES             Location/Qualifiers
     source          1..4538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    53..74
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(89..94,95..112,113..119)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             2396..2413
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2733..3392)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3307..3326
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3393..3495)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      3715..4303
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4204..4223
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4457..4474
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
       61 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      121 gtggaattgt gagcgggata acaatttcac acaggatctc gacatgaaca ataacgatct
      181 ctttcaggca tcacgtcggc gttttctggc acaactcggc ggcttaaccg tcgccgggat
      241 gctggggccg tcattgctaa cgccgcgacg tgcgactgcg catatgccga ttgcgacctc
      301 aacgggtgat aatgttctgg actttacggt tctgggcctg aatagcggta cgagtatgga
      361 tggtattgac tgcgcactgt gtcatttcta tcagaaaacc ccggatgctc cgatggaatt
      421 tgaactgctg gaatacggcg aagttccgct ggcgcagccg attaaacaac gtgtcatgcg
      481 catgatcctg gaagatacca cgagcccgtc tgaactgtca gaagtcaacg tgattctggg
      541 tgaacatttt gcggatgccg tccgtcagtt cgcggccgaa cgcaatgtgg atctgtcaac
      601 cattgacgca atcgcttcgc acggccagac gatttggctg ctgagtatgc cggaagaagg
      661 tcaagtgaaa tccgccctga ccatggcaga aggcgctatc ctggcgagtc gtacgggtat
      721 tacctccatc acggatttcc gtatttccga ccaggcagct ggtcgtcaag gtgcaccgct
      781 gatcgcattt ttcgatgctc tgctgctgca tcacccgacc aaactgcgcg cgtgccagaa
      841 cattggcggt atcgccaatg tgtgttttat tccgccggat gttgacggcc gtcgcaccga
      901 tgaatattac gattttgaca cgggtccggg caacgtgttc atcgacgcag tggttcgtca
      961 ttttaccaat ggtgaacagg aatatgataa agacggtgct atgggcaaac gcggtaaagt
     1021 cgatcaggaa ctggtggatg actttctgaa aatgccgtat ttccaactgg acccgccgaa
     1081 aaccacgggc cgtgaagttt ttcgcgatac cctggcacat gacctgattc gtcgcgcgga
     1141 agccaaaggt ctgagcccgg atgacatcgt ggccaccacg acccgtatta cggcacaggc
     1201 tatcgttgat cactatcgtc gctacgcgcc gtcacaagaa attgacgaaa tcttcatgtg
     1261 cggcggtggc gcctataacc cgaatattgt ggaatttatc cagcaatcgt acccgaacac
     1321 caaaattatg atgctggatg aagcaggtgt cccggcaggt gcaaaagaag cgattacgtt
     1381 cgcctggcag ggcatggaag ccctggttgg tcgtagcatc ccggttccga cccgtgtcga
     1441 aacgcgccag cactatgtgc tgggcaaagt tagcccgggt ctgaattacc gctctgtgat
     1501 gaaaaaaggc atggcatttg gtggcgatgc tcagcaactg ccgtgggttt ctgaaatgat
     1561 cgtgaagaaa aaaggcaaag ttatcaccaa caactgggcg ctcgagcatc cggagaccct
     1621 ggttaaagtc aaagacgcgg aggaccaact gggcgcgcgt gttggctata ttgagctgga
     1681 cctgaattct ggcaaaattc tggaaagctt ccgtccggag gagcgcttcc cgatgatgtc
     1741 taccttcaaa gtgctgctgt gcggtgccgt cctgagccgc gtggatgcgg gtcaggaaca
     1801 gctgggccgt cgtattcatt acagccaaaa cgatctggtg gaatatagcc cggtaaccga
     1861 gaaacacctg accgacggta tgaccgtccg tgagctgtgt agcgcggcga ttacgatgag
     1921 cgacaatacc gccgcgaatc tgctgctgac caccattggc ggcccaaaag aactgacggc
     1981 gttcctgcat aatatgggtg accacgtcac ccgtctggac cgctgggagc cagaactgaa
     2041 cgaggcgatt ccgaatgatg aacgcgatac gaccatgccg gcggcgatgg cgaccaccct
     2101 gcgtaaactg ctgaccggtg agctgctgac cctggcgagc cgccagcagc tgattgattg
     2161 gatggaagcc gacaaagtgg cgggcccgct gctgcgtagc gcgctgccag cgggttggtt
     2221 catcgcggac aaaagcggcg cgggcgaacg cggtagccgt ggcattatcg cggcgctggg
     2281 tccggacggc aaaccgagcc gcattgtcgt gatttatacc accggtagcc aagcgaccat
     2341 ggacgagcgt aaccgtcaaa ttgcggaaat tggcgcgagc ctgatcaaac actggcacca
     2401 tcaccatcac cattaataac ctcagctaag atttttccaa tgatgagcac tttttgatca
     2461 gaaaaaaagg atcatatcgt caattattac ctccacgggg agagcctgag caaactggcc
     2521 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     2581 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt
     2641 gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta gcaaccaggc
     2701 gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag
     2761 tactgttgta attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga
     2821 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg
     2881 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     2941 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
     3001 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     3061 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     3121 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acgtaattcc
     3181 ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta
     3241 tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta
     3301 cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca
     3361 acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc
     3421 gacaactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct
     3481 cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca
     3541 acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg
     3601 gtcgaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3661 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3721 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3781 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3841 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3901 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3961 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4021 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4081 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4141 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4201 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4261 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4321 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4381 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4441 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     4501 accgcctctc cccgcgcgtt ggccgattca ttaatgca
//
