LOCUS       sequence-193810-        3336 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 pLas-tetO combinatorial 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3336)
  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 3336)
  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: pSG81-pLas-tetO-deGFPssrA
FEATURES             Location/Qualifiers
     source          1..3336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    83..101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    121..139
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(264..285)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(525..544)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     872..893
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1109..1126)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1280..1868)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1360..1379)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2039..2830,2831..2899))
                     /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     2662..2681
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2900..3004)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3072..3090
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcattgctgt tccgctgggc atgcttcgag cctagcaagg gtccgggttc accgaaatct
       61 atctcatttg ctagttataa aatccctatc agtgatagag attatgaaat ttgcgtaaat
      121 tccctatcag tgatagagat tcagaagcgg gcccaagttc acttaaaaag gagatcaaca
      181 atgaaagcaa ttttcgtact gaaacatctt aatcatgctg gggagggttt ctaatggagc
      241 ttttcactgg cgttgttccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      301 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      361 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
      421 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      481 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      541 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      601 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
      661 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
      721 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
      781 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
      841 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
      901 ccgggatcgc agcaaacgac gaaaactacg ctttagctgc ttaatgaact caaagcccgc
      961 cgaaaggcgg gcttttctgt ccttgagagt cgggcattgt cttcgctcct tccggtgggc
     1021 gcggggcatg actatcgtcg ccgcacttat gactgtgttc tttatcatgc aactcgtagg
     1081 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1141 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1201 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1261 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1321 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1381 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1441 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1501 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1561 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1621 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1681 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     1741 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1801 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1861 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1921 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     1981 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2041 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2101 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2161 gtgctgcaat gataccgcgt gacccacgct caccggctcc agatttatca gcaataaacc
     2221 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2281 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2341 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2401 gctccggttc ccaacgatca aggcgagttg catgatcccc catgttgtgc aaaaaagcgg
     2461 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     2521 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     2581 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2641 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     2701 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     2761 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     2821 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     2881 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     2941 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3001 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     3061 taacctataa aaataggcgt atcacgaggc cctttcgtgt tcaagaattc tggcgaatcc
     3121 tctgaccagc cagaaaacga cctttctgtg gtgaaaccgg atgctgcaat tcagagcgcc
     3181 agcaagtggg ggacagcaga tgacctgacc gccgcagagt ggatgtttga catggtgatg
     3241 actatcgcac catcagccag aaaaccgaat tttgctgggt gggctaacga tatccgcctg
     3301 atgcgtgaac gtgacggacg taacgaagac aaccac
//
