LOCUS       sequence-225423-        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"
     RBS             345..367
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(404..425)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(665..684)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1012..1033
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1228..1245)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1399..1987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1479..1498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2158..2949,2950..3018))
                     /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     2781..2800
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3019..3123)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3191..3202,1..7)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 acgaggccct ttcgtcttca agaattctgg cgaatcctct gaccagccag aaaacgacct
       61 ttctgtggtg aaaccggatg ctgcaattca gagcggcagc aagtggggga cagcagaaga
      121 cctgaccgcc gcagagtgga tgtttgacat ggtgaagact atcgcaccat cagccagaaa
      181 accgaatttt gctgggtggg ctaacgatat ccgcctgatg cgtgaacgtg acggacgtaa
      241 ccaccgcgac atgtgtgtgc tgttccgctg ggcatgctga gctaacaccg tgcgtgttga
      301 caattttacc tctggcggtg ataatggttg cagctagcaa taattttgtt taactttaag
      361 aaggagatat accatggagc ttttcactgg cgttgttccc atcctggtcg agctggacgg
      421 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      481 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      541 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
      601 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      661 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      721 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
      781 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
      841 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
      901 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
      961 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1021 gctggagttc gtgaccgccg ccgggatcta actcgagcaa agcccgccga aaggcgggct
     1081 tttctgtgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg
     1141 cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga
     1201 caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1261 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1321 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1381 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1441 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1501 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1561 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     1621 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1681 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1741 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1801 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     1861 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1921 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1981 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2041 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2101 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2161 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2221 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2281 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2341 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2401 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2461 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2521 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2581 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2641 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2701 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2761 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2821 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2881 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2941 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3001 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3061 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3121 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     3181 aacctataaa aataggcgta tc
//
