LOCUS       sequence-231641-        3202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses deGFP from a strong sigma 70 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3202)
  AUTHORS   Collias D, Marshall R, Collins SP, Beisel CL, Noireaux V
  TITLE     An educational module to explore CRISPR technologies with a 
            cell-free transcription-translation system.
  JOURNAL   Synth Biol (Oxf). 2019 Jan 21;4(1):ysz005. doi: 
            10.1093/synbio/ysz005. eCollection 2019.
  PUBMED    32995532
REFERENCE   2  (bases 1 to 3202)
  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: "Synth Biol 
            (Oxf)."; date: "2019-01-21"; volume: "4(1):ysz005. doi"; pages: " 
            10.1093/synbio/ysz005. eCollection 2019"
COMMENT     Sequence Label: pColE1_70a_deGFP_AmpR
FEATURES             Location/Qualifiers
     source          1..3202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..47
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             385..407
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(444..465)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(705..724)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1052..1073
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1268..1285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1439..2027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1519..1538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2198..2989,2990..3058))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2821..2840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3059..3163)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
       61 agaattctgg cgaatcctct gaccagccag aaaacgacct ttctgtggtg aaaccggatg
      121 ctgcaattca gagcgccagc aagtggggga cagcagaaga cctgaccgcc gcagagtgga
      181 tgtttgacat ggtgaagact atcgcaccat cagccagaaa accgaatttt gctgggtggg
      241 ctaacgatat ccgcctgatg cgtgaacgtg acggacgtaa ccaccgcgac atgtgtgtgc
      301 tgttccgctg ggcatgctga gctaacaccg tgcgtgttga caattttacc tctggcggtg
      361 ataatggttg cagctagcaa taattttgtt taactttaag aaggagatat accatggagc
      421 ttttcactgg cgttgttccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      481 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      541 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
      601 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      661 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      721 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      781 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
      841 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
      901 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
      961 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1021 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1081 ccgggatcta actcgagcaa agcccgccga aaggcgggct tttctgtgtc gaccgatgcc
     1141 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     1201 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt
     1261 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1321 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1381 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1441 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1501 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1561 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1621 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1681 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1741 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1801 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1861 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1921 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1981 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2041 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2101 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2161 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2221 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2281 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2341 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2401 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2461 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2521 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2581 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2641 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2701 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2761 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg
     2821 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2881 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2941 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3001 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3061 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3121 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3181 tgccacctga cgtctaagaa ac
//
