LOCUS       sequence-154224-        4013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSALECT plasmid with TEM-1 beta-lactamase fused to TEM-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4013)
  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 4013)
  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-TEM1(S70A,D179G)/csTEM1
FEATURES             Location/Qualifiers
     source          1..4013
                     /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             284..1075
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="MHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMM
                     ATFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAIT
                     MSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERGTTMPAAMA
                     TTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGII
                     AALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     primer_bind     complement(436..455)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             1871..1888
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2208..2867)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2782..2801
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2868..2970)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      3190..3778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3679..3698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3932..3949
                     /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 catatgcacc cagaaacgct
      301 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
      361 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatggc
      421 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
      481 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
      541 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
      601 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
      661 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
      721 tgaagccata ccaaacgacg agcgtggcac cacgatgcct gcagcaatgg caacaacgtt
      781 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
      841 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
      901 tattgctgat aaatctggag caggtgagcg tgggtctcgc ggtatcattg cagcactggg
      961 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     1021 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggctcga
     1081 gcatccggag accctggtta aagtcaaaga cgcggaggac caactgggcg cgcgtgttgg
     1141 ctatattgag ctggacctga attctggcaa aattctggaa agcttccgtc cggaggagcg
     1201 cttcccgatg atgtctacct tcaaagtgct gctgtgcggt gccgtcctga gccgcgtgga
     1261 tgcgggtcag gaacagctgg gccgtcgtat tcattacagc caaaacgatc tggtggaata
     1321 tagcccggta accgagaaac acctgaccga cggtatgacc gtccgtgagc tgtgtagcgc
     1381 ggcgattacg atgagcgaca ataccgccgc gaatctgctg ctgaccacca ttggcggccc
     1441 aaaagaactg acggcgttcc tgcataatat gggtgaccac gtcacccgtc tggaccgctg
     1501 ggagccagaa ctgaacgagg cgattccgaa tgatgaacgc gatacgacca tgccggcggc
     1561 gatggcgacc accctgcgta aactgctgac cggtgagctg ctgaccctgg cgagccgcca
     1621 gcagctgatt gattggatgg aagccgacaa agtggcgggc ccgctgctgc gtagcgcgct
     1681 gccagcgggt tggttcatcg cggacaaaag cggcgcgggc gaacgcggta gccgtggcat
     1741 tatcgcggcg ctgggtccgg acggcaaacc gagccgcatt gtcgtgattt ataccaccgg
     1801 tagccaagcg accatggacg agcgtaaccg tcaaattgcg gaaattggcg cgagcctgat
     1861 caaacactgg caccatcacc atcaccatta ataacctcag ctaagatttt tccaatgatg
     1921 agcacttttt gatcagaaaa aaaggatcat atcgtcaatt attacctcca cggggagagc
     1981 ctgagcaaac tggcctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa
     2041 taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac
     2101 gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag
     2161 gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc
     2221 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     2281 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     2341 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     2401 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     2461 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     2521 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     2581 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     2641 gccatacgta attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     2701 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     2761 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     2821 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     2881 gctcctgaaa atctcgacaa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     2941 tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag
     3001 ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc
     3061 acaggtattt attcggtcga aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3121 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3181 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3241 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3301 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3361 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3421 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3481 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3541 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3601 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3661 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3721 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3781 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3841 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3901 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3961 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gca
//
