LOCUS       sequence-181440-        3709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP reporter plasmid with lac operator in its promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3709)
  AUTHORS   Copeland MF, Politz MC, Johnson CB, Markley AL, Pfleger BF
  TITLE     A transcription activator-like effector (TALE) induction system 
            mediated by proteolysis.
  JOURNAL   Nat Chem Biol. 2016 Apr;12(4):254-60. doi: 10.1038/nchembio.2021. 
            Epub 2016 Feb 8.
  PUBMED    26854666
REFERENCE   2  (bases 1 to 3709)
  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: 
            "10.1038/nchembio.2021"; journalName: "Nat Chem Biol"; date: 
            "2016-04"; volume: "12"; issue: "4"; pages: "254-60"
COMMENT     Sequence Label: p5sfGFP
FEATURES             Location/Qualifiers
     source          1..3709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..717)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1..714
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(31..59)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     654..674
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(786..803)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      956..1042
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1134..1161
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1180..1271
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1272..1340,1341..2132)
                     /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     complement(1490..1509)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2303..2891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2792..2811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3045..3062
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3077..3217
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3143..3162)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3303..3325
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        3608..3642
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     protein_bind    3645..3661
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
ORIGIN
        1 atgagcaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
       61 gatgttaatg ggcacaaatt ttctgtccgt ggagagggtg aaggtgatgc tacaaacgga
      121 aaactcaccc ttaaatttat ttgcactact ggaaaactac ctgttccgtg gccaacactt
      181 gtcactactc tgacctatgg tgttcaatgc ttttcccgtt atccggatca catgaaacgg
      241 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaacgcac tatatctttc
      301 aaagatgacg ggacctacaa gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt
      361 aatcgtatcg agttaaaggg tattgatttt aaagaagatg gaaacattct tggacacaaa
      421 cttgagtaca actttaactc acacaatgta tacatcacgg cagacaaaca aaagaatgga
      481 atcaaagcta acttcaaaat tcgccacaac gttgaagatg gttccgttca actagcagac
      541 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
      601 ctgtcgacac aatctgtcct ttcgaaagat cccaacgaaa agcgtgacca catggtcctt
      661 cttgagtttg taactgctgc tgggattaca catggcatgg atgagctcta caaataaggt
      721 acctctagag tcgacctgca ggcatgcaag cttggctgtt ttggcggatg agagaagatt
      781 ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct
      841 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
      901 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
      961 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1021 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
     1081 cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc
     1141 catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac
     1201 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1261 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1321 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1381 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1441 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1501 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1561 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1621 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1681 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1741 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1801 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1861 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1921 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1981 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2041 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2101 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2161 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2221 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2281 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2341 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2401 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     2461 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2521 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2581 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2641 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2701 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2761 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2821 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2881 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2941 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3001 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3061 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3121 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     3181 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     3241 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3301 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca
     3361 gatcaattcg cgcgcgaagg cgaagcggca tgcatttacg ttgcgaattg atctggtttg
     3421 acagcttatc atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag
     3481 ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact
     3541 cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa
     3601 tgagctgttg acagctagct cagtcctagg tactgtgcta gcaattgtga gcggataaca
     3661 atttcgaatt caaaagatct atcgatcgag gatccaggag gtacaatca
//
