LOCUS       sequence-212169-        7544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALEsp2 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 Dec 15, 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_60
FEATURES             Location/Qualifiers
     source          1..7544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        3054..3103
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(3146..3148,3149..3862)
                     /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             3146..3859
                     /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(3176..3204)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3799..3819
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3879..3950
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3966..3993
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    4012..4032
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4033..4090
                     /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(4114..4131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      4665..4887
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4935..5885
                     /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(6511..6605)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6626..7285)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7200..7219
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     7447..7465
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(7469..7512)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    7515..7536
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggagctcggt accaaattcc agaaaagagg cctcccgaaa ggggggcctt ttttcgtttt
       61 ggtcctactg gcgcgcctca gtcagagtat tgacttaaag tctaacctat aggagatcta
      121 cagccatcga gagctgcgag actgtcgccg gatgtgtatc cgacctgacg atggcccaaa
      181 agggccgaaa cagtcctcta caaataattt tgtttaaatc aattcatcga tgtgaaaatg
      241 gtagatttaa gaactttagg atattcacag cagcaacagg aaaagatcaa gcccaaagtt
      301 aggtcgacag tcgcgcagca tcacgaagcg ctggttggtc atgggtttac acatgcccac
      361 atcgtagcct tatcgcagca ccctgcagcc cttggcacgg tcgccgtcaa gtaccaggac
      421 atgattgcgg cgttgccgga agccacacat gaggcgatcg tcggtgtggg gaaacagtgg
      481 agcggagccc gagcgcttga ggccctgttg acggtcgcgg gagagctgag agggcctccc
      541 cttcagctgg acacgggcca gttgctgaag atcgcgaagc ggggaggagt cacggcggtc
      601 gaggcggtgc acgcgtggcg caatgcgctc acgggagcac ccctcaacct gaccccggac
      661 caggtagtcg cgattgcttc acatgacggg ggtaaacaag cgctggaaac ggtgcagcgt
      721 ctgctaccgg tgttatgtca ggatcatggg ctcacgccgg aacaggtagt ggcaattgcg
      781 agtcatgacg gtggcaaaca ggccctggaa accgtacagc ggctgctacc ggtgctgtgt
      841 caagcgcatg gcctgactcc ggaccaagtt gtagccattg cctcgaacgg gggcggcaag
      901 caggcgctgg agactgttca acgtctgctc cccgttctgt gtcaggcgca tggcctgacg
      961 cctgcgcagg tcgtggcgat cgcttcaaac atcggtggga agcaagccct ggagactgtc
     1021 caaagactgt tgccagtgtt gtgtcaagat catggcttaa ccccagatca ggtggttgcg
     1081 attgcatcaa atggaggtgg taaacaggcg ctggagactg tgcagcgcct gttgccggtt
     1141 ctgtgccaag atcatgggct gactccggaa caggttgtgg ctatcgcaag caatattggt
     1201 ggcaagcagg ccctggaaac agtacagcgc ctgctgcctg tattgtgtca agcccacggt
     1261 cttacccccg atcaggtagt cgccatagca tcgcacgacg gcgggaagca ggcccttgag
     1321 actgtacaac gcctcctgcc ggttttgtgc caggcgcacg gcctgacgcc agcccaggtg
     1381 gttgcgatag ccagtaataa cggcggtaag caagcccttg aaaccgttca acgtttgctg
     1441 ccagtgctgt gccaggatca cggcctgacc ccggatcagg tagtcgccat tgctagcaac
     1501 attggtggca aacaagcact ggagacagtt caacgcttac tgcccgtgct ttgccaggat
     1561 catggactga ccccagagca agtggtcgcg attgcctcgc atgacggagg taaacaggcc
     1621 cttgagactg tccagcgtct gctgccggtc ctttgccagg ctcatgggct gacgcccgac
     1681 caggtggtag caatcgcttc gaacggcgga ggcaaacagg cgttagaaac cgttcaacgt
     1741 ctgttaccgg tgctgtgcca agctcatggc ttaaccccgg cccaggtagt cgcaatcgct
     1801 agtcatgatg gtgggaaaca ggcattagaa acagtacaac gtctgctccc ggtcctctgt
     1861 caggatcacg gtctgacccc ggatcaggtt gtagcaatcg ctagcaatat tggtggtaaa
     1921 caggcgctgg aaacagtgca aagattatta ccagttctgt gtcaggacca tggcctgaca
     1981 cctgagcagg tcgtagcgat cgcaagtaat attggaggta aacaggccct ggaaaccgtg
     2041 cagcgtctgc tgcctgtgct ctgtcaagcg catggtctga ctccggatca ggttgtcgcg
     2101 atcgccagta acgggggagg gaaacaggca ctcgagactg tgcagagatt gctgccggtc
     2161 ctgtgtcagg cgcatggtct gaccccagcg caggtcgtgg caatcgcttc caacataggt
     2221 ggtaaacagg ccctcgaaac tgtccaacgg ctgttaccgg tactgtgcca ggatcatggt
     2281 ctgacccctg agcaggtagt ggctattgca tccaacggag ggggcagacc cgcactggag
     2341 tcaatcgtgg cccagctttc gaggccggac cccgcgctgg ccgcactcac taatgatcat
     2401 cttgtagcgc tggcctgcct cggcggacga cccgccttgg atgcggtgaa gaaggggctc
     2461 ccgcacgcgc ctgcattgat taagcggacc aacagaagga ttcccgagag gacatcacat
     2521 cgagtggcag atcacgcgca agtggtccgc gtgctcggat tcttccagtg tcactcccac
     2581 cccgcacaag cgttcgatga cgccatgact caatttggta tgtcgagaca cggactgctg
     2641 cagctctttc gtagagtcgg tgtcacagaa ctcgaggccc gctcgggcac actgcctccc
     2701 gcctcccagc ggtgggacag gattctccaa gcgagcggta tgaaacgcgc gaagccttca
     2761 cctacgtcaa ctcagacacc tgaccaggcg agccttcatg cgttcgcaga ctcgctggag
     2821 agggatttgg acgcgccctc gcccatgcat gaaggggacc aaactcgcgc gtcataatag
     2881 gtttcagcca aaaaacttaa gaccgccggt cttgtccact accttgcagt aatgcggtgg
     2941 acaggatcgg cggttttctt ttctcttctc aagcttatcc cgaaaattta tcaaaaagag
     3001 tattgactta tattgagtcg tataggatac ttacagccat cgagagctgc gagctgtcac
     3061 cggatgtgct ttccggtctg atgagtccgt gaggacgaaa cagcctctac aaataatttt
     3121 gtttaatcac acaggaaagt actagatgag caaaggagaa gaacttttca ctggagttgt
     3181 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tccgtggaga
     3241 gggtgaaggt gatgctacaa acggaaaact cacccttaaa tttatttgca ctactggaaa
     3301 actacctgtt ccgtggccaa cacttgtcac tactctgacc tatggtgttc aatgcttttc
     3361 ccgttatccg gatcacatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta
     3421 tgtacaggaa cgcactatat ctttcaaaga tgacgggacc tacaagacgc gtgctgaagt
     3481 caagtttgaa ggtgataccc ttgttaatcg tatcgagtta aagggtattg attttaaaga
     3541 agatggaaac attcttggac acaaactcga gtacaacttt aactcacaca atgtatacat
     3601 cacggcagac aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacgttga
     3661 agatggttcc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
     3721 tgtcctttta ccagacaacc attacctgtc gacacaatct gtcctttcga aagatcccaa
     3781 cgaaaagcgt gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg
     3841 catggatgag ctctacaaat aatactagag ccaggcatca aataaaacga aaggctcagt
     3901 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
     3961 acactggctc accttcgggt gggcctttct gcgtttatac gctatgtctt ctactagtag
     4021 cggccgctgc agtccggcaa aaaagggcaa ggtgtcacca ccctgccctt tttctttaaa
     4081 accgaaaaga ttacttcgcg ttatgcaggc ttcctcgctc actgactcgc tgcgctcggt
     4141 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatgacag taagacgggt
     4201 aagcctgttg atgataccgc tgccttactg ggtgcattag ccagtctgaa tgacctgtca
     4261 cgggataatc cgaagtggtc agactggaaa atcagagggc aggaactgct gaacagcaaa
     4321 aagtcagata gcaccacata gcagacccgc cataaaacgc cctgagaagc ccgtgacggg
     4381 cttttcttgt attatgggta gtttccttgc atgaatccat aaaaggcgcc tgtagtgcca
     4441 tttaccccca ttcactgcca gagccgtgag cgcagcgaac tgaatgtcac gaaaaagaca
     4501 gcgactcagg tgcctgatgg tcggagacaa aaggaatatt cagcgatttg cccgagcttg
     4561 cgagggtgct acttaagcct ttagggtttt aaggtctgtt ttgtagagga gcaaacagcg
     4621 tttgcgacat ccttttgtaa tactgcggaa ctgactaaag tagtgagtta tacacagggc
     4681 tgggatctat tctttttatc tttttttatt ctttctttat tctataaatt ataaccactt
     4741 gaatataaac aaaaaaaaca cacaaaggtc tagcggaatt tacagagggt ctagcagaat
     4801 ttacaagttt tccagcaaag gtctagcaga atttacagat acccacaact caaaggaaaa
     4861 ggactagtaa ttatcattga ctagcccatc tcaattggta tagtgattaa aatcacctag
     4921 accaattgag atgtatgtct gaattagttg ttttcaaagc aaatgaacta gcgattagtc
     4981 gctatgactt aacggagcat gaaaccaagc taattttatg ctgtgtggca ctactcaacc
     5041 ccacgattga aaaccctaca aggaaagaac ggacggtatc gttcacttat aaccaatacg
     5101 ctcagatgat gaacatcagt agggaaaatg cttatggtgt attagctaaa gcaaccagag
     5161 agctgatgac gagaactgtg gaaatcagga atcctttggt taaaggcttt gagattttcc
     5221 agtggacaaa ctatgccaag ttctcaagcg aaaaattaga attagttttt agtgaagaga
     5281 tattgcctta tcttttccag ttaaaaaaat tcataaaata taatctggaa catgttaagt
     5341 cttttgaaaa caaatactct atgaggattt atgagtggtt attaaaagaa ctaacacaaa
     5401 agaaaactca caaggcaaat atagagatta gccttgatga atttaagttc atgttaatgc
     5461 ttgaaaataa ctaccatgag tttaaaaggc ttaaccaatg ggttttgaaa ccaataagta
     5521 aagatttaaa cacttacagc aatatgaaat tggtggttga taagcgaggc cgcccgactg
     5581 atacgttgat tttccaagtt gaactagata gacaaatgga tctcgtaacc gaacttgaga
     5641 acaaccagat aaaaatgaat ggtgacaaaa taccaacaac cattacatca gattcctacc
     5701 tacataacgg actaagaaaa acactacacg atgctttaac tgcaaaaatt cagctcacca
     5761 gttttgaggc aaaatttttg agtgacatgc aaagtaagta tgatctcaat ggttcgttct
     5821 catggctcac gcaaaaacaa cgaaccacac tagagaacat actggctaaa tacggaagga
     5881 tctgaggttc ttatggctct tgtatctatc agtgaagcat caagactaac aaacaaaagt
     5941 agaacaactg ttcaccgtta catatcaaag ggaaaactgt ccatatgcac agatgaaaac
     6001 ggtgtaaaaa agatagatac atcagagctt ttacgagttt ttggtgcatt caaagctgtt
     6061 caccatgaac agatcgacaa tgtaacagat gaacagcatg taacacctaa tagaacaggt
     6121 gaaaccagta aaacaaagca actagaacat gaaattgaac acctgagaca acttgttaca
     6181 gctcaacagt cacacataga cagcctgaaa caggcgatgc tgcttatcga atcaaagctg
     6241 ccgacaacac gggagccagt gacgcctccc gtggggaaaa aatcatggca attctggaag
     6301 aaatagcgct ttcagccggc aaaccggctg aagccggatc tgcgattctg ataacaaact
     6361 agcaacacca gaacagcccg tttgcgggca gcaaaacccg taccgattat caaaaaggat
     6421 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6481 gtaaacttgg tctgacagct cgaggcttgg attctcacca ataaaaaacg cccggcggca
     6541 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
     6601 gagtccaagc gagctcgata tcaaattacg ccccgccctg ccactcatcg cagtactgtt
     6661 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     6721 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     6781 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     6841 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     6901 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     6961 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     7021 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag
     7081 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     7141 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     7201 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     7261 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     7321 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     7381 gcccgatcaa ctcgagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     7441 tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct
     7501 aaggatgatt tctggaattc gcggccgctt ctagaggaag acac
//
