LOCUS       sequence-197240-        3395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses deGFP fluorescent protein in E. coli. The expression is 
            controlled by a pBAD-tetO combinatorial promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3395)
  AUTHORS   Guo S, Murray RM
  TITLE     Construction of Incoherent Feedforward Loop Circuits in a Cell-Free 
            System and in Cells.
  JOURNAL   ACS Synth Biol. 2019 Mar 15;8(3):606-610. doi: 
            10.1021/acssynbio.8b00493. Epub 2019 Mar 6.
  PUBMED    30790525
REFERENCE   2  (bases 1 to 3395)
  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.8b00493"; journalName: "ACS Synth Biol"; date: 
            "2019-03-15- 15"; volume: "8"; issue: "3"; pages: "606-610"
COMMENT     Sequence Label: pSG15-pBAD-tetO-deGFP
FEATURES             Location/Qualifiers
     source          1..3395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..77
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     protein_bind    155..173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             251..259
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(296..317)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(557..576)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     904..925
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1168..1185)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1339..1927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1419..1438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2098..2889,2890..2958))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHATRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2721..2740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2959..3063)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3131..3149
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcattgctgt tccgctgggc atgcacattg attatttgca cggcgtcaca ctttgctatg
       61 ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta tcgcaactct
      121 ctactgtttc tccataccgt ttttttgggc tagctcccta tcagtgatag agatgcaagc
      181 gggcccaagt tcacttaaaa aggagatcaa caatgaaagc aattttcgta ctgaaacatc
      241 ttaatcatgc taaggaggtt ttctaatgga gcttttcact ggcgttgttc ccatcctggt
      301 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      361 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      421 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      481 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      541 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
      601 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
      661 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
      721 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
      781 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
      841 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
      901 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc taatgaactt tcagccaaaa
      961 aacttaagac cgccggtctt gtccactacc ttgcagtaat gcggtggaca ggatcggcgg
     1021 ttttcttttc tcttctcaac cttgagagtc gggcattgtc ttcgctcctt ccggtgggcg
     1081 cggggcatga ctatcgtcgc cgcacttatg actgtgttct ttatcatgca actcgtagga
     1141 caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1201 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1261 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1321 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1381 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1441 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1501 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1561 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1621 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1681 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1741 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1801 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1861 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1921 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1981 acgttaatgg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2041 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2101 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2161 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2221 tgctgcaatg ataccgcgtg acccacgctc accggctcca gatttatcag caataaacca
     2281 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2341 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2401 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2461 ctccggttcc caacgatcaa ggcgagttgc atgatccccc atgttgtgca aaaaagcggt
     2521 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2581 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2641 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2701 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2761 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2821 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2881 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2941 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3001 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3061 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     3121 aacctataaa aataggcgta tcacgaggcc ctttcgtgtt caagaattct ggcgaatcct
     3181 ctgaccagcc agaaaacgac ctttctgtgg tgaaaccgga tgctgcaatt cagagcgcca
     3241 gcaagtgggg gacagcagat gacctgaccg ccgcagagtg gatgtttgac atggtgatga
     3301 ctatcgcacc atcagccaga aaaccgaatt ttgctgggtg ggctaacgat atccgcctga
     3361 tgcgtgaacg tgacggacgt aacgaagaca accac
//
