LOCUS       sequence-125445-        3799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Derived from 21 with "ClpX" coding sequence from Genbank U00096.3 
            substituted for "deGFP" – used as T500 template for 134-137..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3799)
  AUTHORS   Sun ZZ, Yeung E, Hayes CA, Noireaux V, Murray RM
  TITLE     Linear DNA for rapid prototyping of synthetic biological circuits in
            an Escherichia coli based TX-TL cell-free system.
  JOURNAL   ACS Synth Biol. 2014 Jun 20;3(6):387-97. doi: 10.1021/sb400131a. 
            Epub 2013 Dec 4.
  PUBMED    24303785
REFERENCE   2  (bases 1 to 3799)
  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/sb400131a";
            journalName: "ACS Synth Biol"; date: "2014-06-20- 20"; volume: "3"; 
            issue: "6"; pages: "387-97"
COMMENT     Sequence Label: pBEST_OR2-OR1-Pr-UTR1_ClpX-T500 (48p)
FEATURES             Location/Qualifiers
     source          1..3799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             7..29
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1487..1504)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1658..2246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1738..1757)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2417..3208,3209..3277))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3040..3059
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3278..3382)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3450..3468
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aataattttg tttaacttta agaaggagat ataccatgac agataaacgc aaagatggct
       61 caggcaaatt gctgtattgc tctttttgcg gcaaaagcca gcatgaagtg cgcaagctga
      121 ttgccggtcc atccgtgtat atctgcgacg aatgtgttga tttatgtaac gacatcattc
      181 gcgaagagat taaagaagtt gcaccgcatc gtgaacgcag tgcgctaccg acgccgcatg
      241 aaattcgcaa ccacctggac gattacgtta tcggccagga acaggcgaaa aaagtgctgg
      301 cggtcgcggt atacaaccat tacaaacgtc tgcgcaacgg cgataccagc aatggcgtcg
      361 agttgggcaa aagtaacatt ctgctgatcg gtccgaccgg ttccggtaaa acgctgctgg
      421 ctgaaacgct ggcgcgcctg ctggatgttc cgttcaccat ggccgacgcg actacactga
      481 ccgaagccgg ttatgtgggt gaagacgttg aaaacatcat tcagaagctg ttgcagaaat
      541 gcgactacga tgtccagaaa gcacagcgtg gtattgtcta catcgatgaa atcgacaaga
      601 tttctcgtaa gtcagacaac ccgtccatta cccgagacgt ttccggtgaa ggcgtacagc
      661 aggcactgtt gaaactgatc gaaggtacgg tagctgctgt tccaccgcaa ggtgggcgta
      721 aacatccgca gcaggaattc ttgcaggttg atacctctaa gatcctgttt atttgtggcg
      781 gtgcgtttgc cggtctggat aaagtgattt cccaccgtgt agaaaccggc tccggcattg
      841 gttttggcgc gacggtaaaa gcgaagtccg acaaagcaag cgaaggcgag ctgctggcgc
      901 aggttgaacc ggaagatctg atcaagtttg gtcttatccc tgagtttatt ggtcgtctgc
      961 cggttgtcgc aacgttgaat gaactgagcg aagaagctct gattcagatc ctcaaagagc
     1021 cgaaaaacgc cctgaccaag cagtatcagg cgctgtttaa tctggaaggc gtggatctgg
     1081 aattccgtga cgaggcgctg gatgctatcg ctaagaaagc gatggcgcgt aaaaccggtg
     1141 cccgtggcct gcgttccatc gtagaagccg cactgctcga taccatgtac gatctgccgt
     1201 ccatggaaga cgtcgaaaaa gtggttatcg acgagtcggt aattgatggt caaagcaaac
     1261 cgttgctgat ttatggcaag ccggaagcgc aacaggcatc tggtgaataa ctcgagcaaa
     1321 gcccgccgaa aggcgggctt ttctgtgtcg accgatgccc ttgagagcct tcaacccagt
     1381 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     1441 tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact
     1501 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1561 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1621 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1681 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1741 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1801 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     1861 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1921 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1981 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2041 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     2101 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2161 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2221 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2281 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2341 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2401 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2461 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2521 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     2581 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2641 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     2701 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     2761 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     2821 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     2881 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2941 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3001 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3061 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3121 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3181 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3241 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3301 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3361 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     3421 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     3481 aagaattctg gcgaatcctc tgaccagcca gaaaacgacc tttctgtggt gaaaccggat
     3541 gctgcaattc agagcggcag caagtggggg acagcagaag acctgaccgc cgcagagtgg
     3601 atgtttgaca tggtgaagac tatcgcacca tcagccagaa aaccgaattt tgctgggtgg
     3661 gctaacgata tccgcctgat gcgtgaacgt gacggacgta accaccgcga catgtgtgtg
     3721 ctgttccgct gggcatgctg agctaacacc gtgcgtgttg acaattttac ctctggcggt
     3781 gataatggtt gcagctagc
//
