LOCUS       sequence-193809-        3428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses deGFP fluorescent protein with ssrA degradation tag 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 3428)
  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 3428)
  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: pSG25-pBAD-tetO-deGFPssrA
FEATURES             Location/Qualifiers
     source          1..3428
                     /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(1201..1218)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1372..1960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1452..1471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2131..2922,2923..2991))
                     /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     2754..2773
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2992..3096)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3164..3182
                     /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 gcagcaaacg acgaaaacta
      961 cgctttagct gcttaatgaa ctttcagcca aaaaacttaa gaccgccggt cttgtccact
     1021 accttgcagt aatgcggtgg acaggatcgg cggttttctt ttctcttctc aaccttgaga
     1081 gtcgggcatt gtcttcgctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     1141 atgactgtgt tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc
     1201 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1261 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1321 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1381 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1441 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1501 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1561 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1621 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1681 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1741 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1801 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1861 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1921 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1981 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2041 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2101 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2161 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2221 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gtgacccacg
     2281 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2341 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2401 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2461 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2521 tgcatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2581 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2641 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2701 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac
     2761 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2821 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2881 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2941 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3001 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3061 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3121 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     3181 gccctttcgt gttcaagaat tctggcgaat cctctgacca gccagaaaac gacctttctg
     3241 tggtgaaacc ggatgctgca attcagagcg ccagcaagtg ggggacagca gatgacctga
     3301 ccgccgcaga gtggatgttt gacatggtga tgactatcgc accatcagcc agaaaaccga
     3361 attttgctgg gtgggctaac gatatccgcc tgatgcgtga acgtgacgga cgtaacgaag
     3421 acaaccac
//
