LOCUS       sequence-248648-        2868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes crippled human pEF1alpha as a Type 2 part to be used in the 
            MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2868)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells.
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 2868)
  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: 
            "10.1021/acssynbio.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: MTK2_025
FEATURES             Location/Qualifiers
     source          1..2868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     45..64
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      312..900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     801..820
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1020..2198
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1251..2189
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2146..2166
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     terminator      complement(2234..2268)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(join(2339..2868,1..130))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
       61 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
      121 ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
      181 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgcccg atcaatcatg
      241 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
      301 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
      361 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      421 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
      481 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      541 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      601 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      661 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      721 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      781 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      841 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      901 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      961 ttctttcctg cgttatcccc tgattctgtg gataaccgta gtcggtctca aacgcgtgag
     1021 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     1081 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     1141 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     1201 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
     1261 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
     1321 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
     1381 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
     1441 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
     1501 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
     1561 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
     1621 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
     1681 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
     1741 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
     1801 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga
     1861 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
     1921 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
     1981 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     2041 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     2101 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     2161 tggttcaaag tttttttctt ccatttcagg tgtcgtgaag atcgccgccg cagatctatg
     2221 tgagaccaga ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     2281 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
     2341 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     2401 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     2461 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     2521 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgaaacgaaa aacatattct
     2581 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     2641 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     2701 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     2761 cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa
     2821 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaa
//
