LOCUS       sequence-206007-        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        3062..3111
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(3154..3156,3157..3870)
                     /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             3154..3867
                     /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(3184..3212)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3807..3827
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3887..3958
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3974..4001
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    4020..4040
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4041..4098
                     /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(4122..4139)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      4673..4895
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4943..5893
                     /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(6519..6613)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6634..7293)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7208..7227
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     7455..7473
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(7477..7520)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    7523..7544
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 gaagacacgg agctcggtac caaattccag aaaagaggcc tcccgaaagg ggggcctttt
       61 ttcgttttgg tcctactggc gcgcctcagt cagagtattg acttaaagtc taacctatag
      121 gagatctaca gccatcgaga gctgcgagac tgtcgccgga tgtgtatccg acctgacgat
      181 ggcccaaaag ggccgaaaca gtcctctaca aataattttg tttaaatcaa ttcatcgacg
      241 tgaaaatggt agatttaaga actttaggat attcacagca gcaacaggaa aagatcaagc
      301 ccaaagttag gtcgacagtc gcgcagcatc acgaagcgct ggttggtcat gggtttacac
      361 atgcccacat cgtagcctta tcgcagcacc ctgcagccct tggcacggtc gccgtcaagt
      421 accaggacat gattgcggcg ttgccggaag ccacacatga ggcgatcgtc ggtgtgggga
      481 aacagtggag cggagcccga gcgcttgagg ccctgttgac ggtcgcggga gagctgagag
      541 ggcctcccct tcagctggac acgggccagt tgctgaagat cgcgaagcgg ggaggagtca
      601 cggcggtcga ggcggtgcac gcgtggcgca atgcgctcac gggagcaccc ctcaacctga
      661 ccccggacca ggtagtcgcg attgcttcac atgacggggg taaacaagcg ctggaaacgg
      721 tgcagcgtct gctaccggtg ttatgtcagg atcatgggct cacgccggaa caggtagtgg
      781 caattgcgag tcatgacggt ggcaaacagg ccctggaaac cgtacagcgg ctgctaccgg
      841 tgctgtgtca agcgcatggc ctgactccgg accaagttgt agccattgcc tcgaacgggg
      901 gcggcaagca ggcgctggag actgttcaac gtctgctccc cgttctgtgt caggcgcatg
      961 gcctgacgcc tgcgcaggtc gtggcgatcg cttcaaacat cggtgggaag caagccctgg
     1021 agactgtcca aagactgttg ccagtgttgt gtcaagatca tggcttaacc ccagatcagg
     1081 tggttgcgat tgcatcaaat ggaggtggta aacaggcgct ggagactgtg cagcgcctgt
     1141 tgccggttct gtgccaagat catgggctga ctccggaaca ggttgtggct atcgcaagca
     1201 atattggtgg caagcaggcc ctggaaacag tacagcgcct gctgcctgta ttgtgtcaag
     1261 cccacggtct tacccccgat caggtagtcg ccatagcatc gcacgacggc gggaagcagg
     1321 cccttgagac tgtacaacgc ctcctgccgg ttttgtgcca ggcgcacggc ctgacgccag
     1381 cccaggtggt tgcgatagcc agtaataacg gcggtaagca agcccttgaa accgttcaac
     1441 gtttgctgcc agtgctgtgc caggatcacg gcctgacccc ggatcaggta gtcgccattg
     1501 ctagcaacat tggtggcaaa caagcactgg agacagttca acgcttactg cccgtgcttt
     1561 gccaggatca tggactgacc ccagagcaag tggtcgcgat tgcctcgcat gacggaggta
     1621 aacaggccct tgagactgtc cagcgtctgc tgccggtcct ttgccaggct catgggctga
     1681 cgcccgacca ggtggtagca atcgcttcga acggcggagg caaacaggcg ttagaaaccg
     1741 ttcaacgtct gttaccggtg ctgtgccaag ctcatggctt aaccccggcc caggtagtcg
     1801 caatcgctag tcatgatggt gggaaacagg cattagaaac agtacaacgt ctgctcccgg
     1861 tcctctgtca ggatcacggt ctgaccccgg atcaggttgt agcaatcgct agcaatattg
     1921 gtggtaaaca ggcgctggaa acagtgcaaa gattattacc agttctgtgt caggaccatg
     1981 gcctgacacc tgagcaggtc gtagcgatcg caagtaatat tggaggtaaa caggccctgg
     2041 aaaccgtgca gcgtctgctg cctgtgctct gtcaagcgca tggtctgact ccggatcagg
     2101 ttgtcgcgat cgccagtaac gggggaggga aacaggcact cgagactgtg cagagattgc
     2161 tgccggtcct gtgtcaggcg catggtctga ccccagcgca ggtcgtggca atcgcttcca
     2221 acataggtgg taaacaggcc ctcgaaactg tccaacggct gttaccggta ctgtgccagg
     2281 atcatggtct gacccctgag caggtagtgg ctattgcatc caacggaggg ggcagacccg
     2341 cactggagtc aatcgtggcc cagctttcga ggccggaccc cgcgctggcc gcactcacta
     2401 atgatcatct tgtagcgctg gcctgcctcg gcggacgacc cgccttggat gcggtgaaga
     2461 aggggctccc gcacgcgcct gcattgatta agcggaccaa cagaaggatt cccgagagga
     2521 catcacatcg agtggcagat cacgcgcaag tggtccgcgt gctcggattc ttccagtgtc
     2581 actcccaccc cgcacaagcg ttcgatgacg ccatgactca atttggtatg tcgagacacg
     2641 gactgctgca gctctttcgt agagtcggtg tcacagaact cgaggcccgc tcgggcacac
     2701 tgcctcccgc ctcccagcgg tgggacagga ttctccaagc gagcggtatg aaacgcgcga
     2761 agccttcacc tacgtcaact cagacacctg accaggcgag ccttcatgcg ttcgcagact
     2821 cgctggagag ggatttggac gcgccctcgc ccatgcatga aggggaccaa actcgcgcgt
     2881 cataataggt ttcagccaaa aaacttaaga ccgccggtct tgtccactac cttgcagtaa
     2941 tgcggtggac aggatcggcg gttttctttt ctcttctcaa gcttatcccg aaaatttatc
     3001 aaaaagagta ttgacttata ttgagtcgta taggatactt acagccatcg agagctgcga
     3061 gctgtcaccg gatgtgcttt ccggtctgat gagtccgtga ggacgaaaca gcctctacaa
     3121 ataattttgt ttaatcacac aggaaagtac tagatgagca aaggagaaga acttttcact
     3181 ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc
     3241 cgtggagagg gtgaaggtga tgctacaaac ggaaaactca cccttaaatt tatttgcact
     3301 actggaaaac tacctgttcc gtggccaaca cttgtcacta ctctgaccta tggtgttcaa
     3361 tgcttttccc gttatccgga tcacatgaaa cggcatgact ttttcaagag tgccatgccc
     3421 gaaggttatg tacaggaacg cactatatct ttcaaagatg acgggaccta caagacgcgt
     3481 gctgaagtca agtttgaagg tgataccctt gttaatcgta tcgagttaaa gggtattgat
     3541 tttaaagaag atggaaacat tcttggacac aaactcgagt acaactttaa ctcacacaat
     3601 gtatacatca cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattcgccac
     3661 aacgttgaag atggttccgt tcaactagca gaccattatc aacaaaatac tccaattggc
     3721 gatggccctg tccttttacc agacaaccat tacctgtcga cacaatctgt cctttcgaaa
     3781 gatcccaacg aaaagcgtga ccacatggtc cttcttgagt ttgtaactgc tgctgggatt
     3841 acacatggca tggatgagct ctacaaataa tactagagcc aggcatcaaa taaaacgaaa
     3901 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta
     3961 ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatacgc tatgtcttct
     4021 actagtagcg gccgctgcag tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt
     4081 tctttaaaac cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac tgactcgctg
     4141 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatgacagta
     4201 agacgggtaa gcctgttgat gataccgctg ccttactggg tgcattagcc agtctgaatg
     4261 acctgtcacg ggataatccg aagtggtcag actggaaaat cagagggcag gaactgctga
     4321 acagcaaaaa gtcagatagc accacatagc agacccgcca taaaacgccc tgagaagccc
     4381 gtgacgggct tttcttgtat tatgggtagt ttccttgcat gaatccataa aaggcgcctg
     4441 tagtgccatt tacccccatt cactgccaga gccgtgagcg cagcgaactg aatgtcacga
     4501 aaaagacagc gactcaggtg cctgatggtc ggagacaaaa ggaatattca gcgatttgcc
     4561 cgagcttgcg agggtgctac ttaagccttt agggttttaa ggtctgtttt gtagaggagc
     4621 aaacagcgtt tgcgacatcc ttttgtaata ctgcggaact gactaaagta gtgagttata
     4681 cacagggctg ggatctattc tttttatctt tttttattct ttctttattc tataaattat
     4741 aaccacttga atataaacaa aaaaaacaca caaaggtcta gcggaattta cagagggtct
     4801 agcagaattt acaagttttc cagcaaaggt ctagcagaat ttacagatac ccacaactca
     4861 aaggaaaagg actagtaatt atcattgact agcccatctc aattggtata gtgattaaaa
     4921 tcacctagac caattgagat gtatgtctga attagttgtt ttcaaagcaa atgaactagc
     4981 gattagtcgc tatgacttaa cggagcatga aaccaagcta attttatgct gtgtggcact
     5041 actcaacccc acgattgaaa accctacaag gaaagaacgg acggtatcgt tcacttataa
     5101 ccaatacgct cagatgatga acatcagtag ggaaaatgct tatggtgtat tagctaaagc
     5161 aaccagagag ctgatgacga gaactgtgga aatcaggaat cctttggtta aaggctttga
     5221 gattttccag tggacaaact atgccaagtt ctcaagcgaa aaattagaat tagtttttag
     5281 tgaagagata ttgccttatc ttttccagtt aaaaaaattc ataaaatata atctggaaca
     5341 tgttaagtct tttgaaaaca aatactctat gaggatttat gagtggttat taaaagaact
     5401 aacacaaaag aaaactcaca aggcaaatat agagattagc cttgatgaat ttaagttcat
     5461 gttaatgctt gaaaataact accatgagtt taaaaggctt aaccaatggg ttttgaaacc
     5521 aataagtaaa gatttaaaca cttacagcaa tatgaaattg gtggttgata agcgaggccg
     5581 cccgactgat acgttgattt tccaagttga actagataga caaatggatc tcgtaaccga
     5641 acttgagaac aaccagataa aaatgaatgg tgacaaaata ccaacaacca ttacatcaga
     5701 ttcctaccta cataacggac taagaaaaac actacacgat gctttaactg caaaaattca
     5761 gctcaccagt tttgaggcaa aatttttgag tgacatgcaa agtaagtatg atctcaatgg
     5821 ttcgttctca tggctcacgc aaaaacaacg aaccacacta gagaacatac tggctaaata
     5881 cggaaggatc tgaggttctt atggctcttg tatctatcag tgaagcatca agactaacaa
     5941 acaaaagtag aacaactgtt caccgttaca tatcaaaggg aaaactgtcc atatgcacag
     6001 atgaaaacgg tgtaaaaaag atagatacat cagagctttt acgagttttt ggtgcattca
     6061 aagctgttca ccatgaacag atcgacaatg taacagatga acagcatgta acacctaata
     6121 gaacaggtga aaccagtaaa acaaagcaac tagaacatga aattgaacac ctgagacaac
     6181 ttgttacagc tcaacagtca cacatagaca gcctgaaaca ggcgatgctg cttatcgaat
     6241 caaagctgcc gacaacacgg gagccagtga cgcctcccgt ggggaaaaaa tcatggcaat
     6301 tctggaagaa atagcgcttt cagccggcaa accggctgaa gccggatctg cgattctgat
     6361 aacaaactag caacaccaga acagcccgtt tgcgggcagc aaaacccgta ccgattatca
     6421 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6481 atatatgagt aaacttggtc tgacagctcg aggcttggat tctcaccaat aaaaaacgcc
     6541 cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct
     6601 atcaacagga gtccaagcga gctcgatatc aaattacgcc ccgccctgcc actcatcgca
     6661 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
     6721 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
     6781 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     6841 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
     6901 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     6961 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     7021 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc
     7081 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     7141 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     7201 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     7261 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     7321 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     7381 tcttacgtgc ccgatcaact cgagtgccac ctgacgtcta agaaaccatt attatcatga
     7441 cattaaccta taaaaatagg cgtatcacga ggcagaattt cagataaaaa aaatccttag
     7501 ctttcgctaa ggatgatttc tggaattcgc ggccgcttct agag
//
