LOCUS       sequence-212168-        7544 bp    DNA     circular SYN 14-APR-2025
DEFINITION  TALEsp1 stabilized promoter driving GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7544)
  AUTHORS   Segall-Shapiro TH, Sontag ED, Voigt CA
  TITLE     Engineered promoters enable constant gene expression at any copy 
            number in bacteria.
  JOURNAL   Nat Biotechnol. 2018 Mar 19. pii: nbt.4111. doi: 10.1038/nbt.4111.
  PUBMED    29553576
REFERENCE   2  (bases 1 to 7544)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2018 Mar 19. pii: nbt.4111. doi: 10.1038/nbt.4111."
COMMENT     Sequence Label: pTHSSe_59
FEATURES             Location/Qualifiers
     source          1..7544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        2459..2508
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(2551..2553,2554..3267)
                     /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             2551..3264
                     /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(2581..2609)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3204..3224
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3284..3355
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3371..3398
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3417..3437
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3438..3495
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(3519..3536)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      4070..4292
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4340..5290
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(5916..6010)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6031..6690)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6605..6624
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     6852..6870
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(6874..6917)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    6920..6941
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 cggtcgaggc ggtgcacgcg tggcgcaatg cgctcacggg agcacccctc aatctgaccc
       61 cggatcaggt ggttgcgatc gccagtaatg gcggaggcaa acaagcactc gaaactgtac
      121 agcgcctcct gccggtactg tgccaagatc atggcttgac gcctgagcaa gtcgtagcta
      181 ttgcatcaaa catcggtggt aaacaggcgc tggaaaccgt acaacgatta ctccctgtct
      241 tatgccaggc acacggtctg acccccgacc aagttgtagc cattgcgtct aatggcggcg
      301 ggaaacaggc cctggaaacg gtccaacgtc tgttacccgt tctgtgtcag gctcacggtc
      361 tgacccctgc ccaggtagtt gcaattgcca gcaacatcgg cgggaaacaa gcgctggaga
      421 ctgtgcagcg tctgctccct gtgttatgcc aagatcatgg gctcactccg gatcaggtgg
      481 tggccatcgc ttccaatatt ggcggtaaac aggcgctgga gacagtgcaa cgacttttac
      541 ctgttctctg ccaggatcat ggtctaactc ccgagcaggt cgtcgccatc gcctctcatg
      601 acggcgggaa acaagcgttg gaaactgtcc agcgactcct gccggttttg tgccaggccc
      661 acgggcttac tcctgaccag gtagttgcga tcgcgtcaaa tgggggtggc aaacaagccc
      721 tcgaaaccgt gcaacgcctg ctgcccgtct tgtgccaagc tcatgggctg actccggcgc
      781 aagtagtcgc gattgcgagc cacgatggcg gtaagcaggc actggaaacg gttcagcgcc
      841 tgctcccggt tctatgccag gatcacggcc tgaccccgga ccaggtcgtc gcgatcgcgt
      901 caaatatcgg tggcaaacaa gctttggaga cagtacagcg cctgttacca gtgctttgcc
      961 aggaccatgg tctgacccct gagcaagtag tggcgatcgc ttctaatatt gggggcaaac
     1021 aagcgctgga aacagtacag cgtctgttac cggtcctatg ccaggcacat ggcctgaccc
     1081 ctgatcaggt ggtagccatt gccagtcatg atggcggtaa acaggcgctt gagactgtcc
     1141 aacgtctgct gccggtcctc tgtcaggctc atggcctgac gccagctcaa gtcgtggcta
     1201 tcgcttcgca tgatggcgga aaacaggcac tggagactgt gcagcgactg ttgccagttc
     1261 tgtgtcagga tcacggttta actccggacc aggtggtcgc tattgcgtcg aatgggggcg
     1321 gtaaacaagc gctggaaact gtgcaacgtt tgctcccagt tctgtgccag gaccatgggc
     1381 tgactccgga acaggtagtg gccattgctt ctaatattgg tgggaaacag gcgctggaaa
     1441 ccgtgcagcg cctgcttcca gtgctttgcc aggcccatgg cctgacgcca gatcaggtgg
     1501 ttgctatagc cagcaatggc ggcggtaaac aggccctcga aaccgtccag cgcctgctcc
     1561 ctgtgctgtg ccaggcccat gggcttaccc cagcgcaagt agtggcgatt gcgtctaata
     1621 ttggtggtaa acaggcgttg gagactgtac aacgcctgct gccagtttta tgccaagatc
     1681 atggtctgac ccctgagcag gtagtggcta ttgcatccaa caacgggggc agacccgcac
     1741 tggagtcaat cgtggcccag ctttcgaggc cggaccccgc gctggccgca ctcactaatg
     1801 atcatcttgt agcgctggcc tgcctcggcg gacgacccgc cttggatgcg gtgaagaagg
     1861 ggctcccgca cgcgcctgca ttgattaagc ggaccaacag aaggattccc gagaggacat
     1921 cacatcgagt ggcagatcac gcgcaagtgg tccgcgtgct cggattcttc cagtgtcact
     1981 cccaccccgc acaagcgttc gatgacgcca tgactcaatt tggtatgtcg agacacggac
     2041 tgctgcagct ctttcgtaga gtcggtgtca cagaactcga ggcccgctcg ggcacactgc
     2101 ctcccgcctc ccagcggtgg gacaggattc tccaagcgag cggtatgaaa cgcgcgaagc
     2161 cttcacctac gtcaactcag acacctgacc aggcgagcct tcatgcgttc gcagactcgc
     2221 tggagaggga tttggacgcg ccctcgccca tgcatgaagg ggaccaaact cgcgcgtcat
     2281 aataggtttc agccaaaaaa cttaagaccg ccggtcttgt ccactacctt gcagtaatgc
     2341 ggtggacagg atcggcggtt ttcttttctc ttctcaagct tatcccgaaa atttatcaaa
     2401 aagagtattg acttataact caacctatag gatacttaca gccatcgaga gctgcgagct
     2461 gtcaccggat gtgctttccg gtctgatgag tccgtgagga cgaaacagcc tctacaaata
     2521 attttgttta atcacacagg aaagtactag atgagcaaag gagaagaact tttcactgga
     2581 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtccgt
     2641 ggagagggtg aaggtgatgc tacaaacgga aaactcaccc ttaaatttat ttgcactact
     2701 ggaaaactac ctgttccgtg gccaacactt gtcactactc tgacctatgg tgttcaatgc
     2761 ttttcccgtt atccggatca catgaaacgg catgactttt tcaagagtgc catgcccgaa
     2821 ggttatgtac aggaacgcac tatatctttc aaagatgacg ggacctacaa gacgcgtgct
     2881 gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg agttaaaggg tattgatttt
     2941 aaagaagatg gaaacattct tggacacaaa ctcgagtaca actttaactc acacaatgta
     3001 tacatcacgg cagacaaaca aaagaatgga atcaaagcta acttcaaaat tcgccacaac
     3061 gttgaagatg gttccgttca actagcagac cattatcaac aaaatactcc aattggcgat
     3121 ggccctgtcc ttttaccaga caaccattac ctgtcgacac aatctgtcct ttcgaaagat
     3181 cccaacgaaa agcgtgacca catggtcctt cttgagtttg taactgctgc tgggattaca
     3241 catggcatgg atgagctcta caaataatac tagagccagg catcaaataa aacgaaaggc
     3301 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
     3361 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatacgctat gtcttctact
     3421 agtagcggcc gctgcagtcc ggcaaaaaag ggcaaggtgt caccaccctg ccctttttct
     3481 ttaaaaccga aaagattact tcgcgttatg caggcttcct cgctcactga ctcgctgcgc
     3541 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat gacagtaaga
     3601 cgggtaagcc tgttgatgat accgctgcct tactgggtgc attagccagt ctgaatgacc
     3661 tgtcacggga taatccgaag tggtcagact ggaaaatcag agggcaggaa ctgctgaaca
     3721 gcaaaaagtc agatagcacc acatagcaga cccgccataa aacgccctga gaagcccgtg
     3781 acgggctttt cttgtattat gggtagtttc cttgcatgaa tccataaaag gcgcctgtag
     3841 tgccatttac ccccattcac tgccagagcc gtgagcgcag cgaactgaat gtcacgaaaa
     3901 agacagcgac tcaggtgcct gatggtcgga gacaaaagga atattcagcg atttgcccga
     3961 gcttgcgagg gtgctactta agcctttagg gttttaaggt ctgttttgta gaggagcaaa
     4021 cagcgtttgc gacatccttt tgtaatactg cggaactgac taaagtagtg agttatacac
     4081 agggctggga tctattcttt ttatcttttt ttattctttc tttattctat aaattataac
     4141 cacttgaata taaacaaaaa aaacacacaa aggtctagcg gaatttacag agggtctagc
     4201 agaatttaca agttttccag caaaggtcta gcagaattta cagataccca caactcaaag
     4261 gaaaaggact agtaattatc attgactagc ccatctcaat tggtatagtg attaaaatca
     4321 cctagaccaa ttgagatgta tgtctgaatt agttgttttc aaagcaaatg aactagcgat
     4381 tagtcgctat gacttaacgg agcatgaaac caagctaatt ttatgctgtg tggcactact
     4441 caaccccacg attgaaaacc ctacaaggaa agaacggacg gtatcgttca cttataacca
     4501 atacgctcag atgatgaaca tcagtaggga aaatgcttat ggtgtattag ctaaagcaac
     4561 cagagagctg atgacgagaa ctgtggaaat caggaatcct ttggttaaag gctttgagat
     4621 tttccagtgg acaaactatg ccaagttctc aagcgaaaaa ttagaattag tttttagtga
     4681 agagatattg ccttatcttt tccagttaaa aaaattcata aaatataatc tggaacatgt
     4741 taagtctttt gaaaacaaat actctatgag gatttatgag tggttattaa aagaactaac
     4801 acaaaagaaa actcacaagg caaatataga gattagcctt gatgaattta agttcatgtt
     4861 aatgcttgaa aataactacc atgagtttaa aaggcttaac caatgggttt tgaaaccaat
     4921 aagtaaagat ttaaacactt acagcaatat gaaattggtg gttgataagc gaggccgccc
     4981 gactgatacg ttgattttcc aagttgaact agatagacaa atggatctcg taaccgaact
     5041 tgagaacaac cagataaaaa tgaatggtga caaaatacca acaaccatta catcagattc
     5101 ctacctacat aacggactaa gaaaaacact acacgatgct ttaactgcaa aaattcagct
     5161 caccagtttt gaggcaaaat ttttgagtga catgcaaagt aagtatgatc tcaatggttc
     5221 gttctcatgg ctcacgcaaa aacaacgaac cacactagag aacatactgg ctaaatacgg
     5281 aaggatctga ggttcttatg gctcttgtat ctatcagtga agcatcaaga ctaacaaaca
     5341 aaagtagaac aactgttcac cgttacatat caaagggaaa actgtccata tgcacagatg
     5401 aaaacggtgt aaaaaagata gatacatcag agcttttacg agtttttggt gcattcaaag
     5461 ctgttcacca tgaacagatc gacaatgtaa cagatgaaca gcatgtaaca cctaatagaa
     5521 caggtgaaac cagtaaaaca aagcaactag aacatgaaat tgaacacctg agacaacttg
     5581 ttacagctca acagtcacac atagacagcc tgaaacaggc gatgctgctt atcgaatcaa
     5641 agctgccgac aacacgggag ccagtgacgc ctcccgtggg gaaaaaatca tggcaattct
     5701 ggaagaaata gcgctttcag ccggcaaacc ggctgaagcc ggatctgcga ttctgataac
     5761 aaactagcaa caccagaaca gcccgtttgc gggcagcaaa acccgtaccg attatcaaaa
     5821 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5881 tatgagtaaa cttggtctga cagctcgagg cttggattct caccaataaa aaacgcccgg
     5941 cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc
     6001 aacaggagtc caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta
     6061 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac
     6121 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa
     6181 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
     6241 ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag
     6301 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
     6361 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
     6421 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg
     6481 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
     6541 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
     6601 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     6661 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     6721 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     6781 tacgtgcccg atcaactcga gtgccacctg acgtctaaga aaccattatt atcatgacat
     6841 taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt
     6901 tcgctaagga tgatttctgg aattcgcggc cgcttctaga ggaagacacg gagctcggta
     6961 ccaaattcca gaaaagaggc ctcccgaaag gggggccttt tttcgttttg gtcctactgg
     7021 cgcgcctcag tcagagtatt gacttaaagt ctaacctata ggaccgttac agccatcgag
     7081 agctgcgaga ctgtcgccgg atgtgtatcc gacctgacga tggcccaaaa gggccgaaac
     7141 agtcctctac aaataatttt gtttaaatca attcacctgc gtgaaaatgg tagatttaag
     7201 aactttagga tattcacagc agcaacagga aaagatcaag cccaaagtta ggtcgacagt
     7261 cgcgcagcat cacgaagcgc tggttggtca tgggtttaca catgcccaca tcgtagcctt
     7321 atcgcagcac cctgcagccc ttggcacggt cgccgtcaag taccaggaca tgattgcggc
     7381 gttgccggaa gccacacatg aggcgatcgt cggtgtgggg aaacagtgga gcggagcccg
     7441 agcgcttgag gccctgttga cggtcgcggg agagctgaga gggcctcccc ttcagctgga
     7501 cacgggccag ttgctgaaga tcgcgaagcg gggaggagtc acgg
//
