LOCUS       sequence-212167-        6897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PTac driving TALEsp1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6897)
  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 6897)
  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_39
FEATURES             Location/Qualifiers
     source          1..6897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    33..52
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     misc_RNA        70..119
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     terminator      2815..2886
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2902..2929
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2936..2956
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2957..3014
                     /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."
     CDS             complement(3172..3963)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPVESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRTDQLEEFVHYVKGEITKVVGK"
     rep_origin      complement(4558..5103)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4638..4657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5571..5589
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5593..5636)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(5679..6761)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6723..6742
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6896..6897,1..4,5..20,21..27)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 gacaattaat catcggctcg tataatgtgt ggaattgtga gcgctcacaa ttctatggac
       61 tatgtttagc tgtcaccgga tgtgctttcc ggtctgatga gtccgtgagg acgaaacagc
      121 ctctacaaat aattttgttt aatcacacag gaaagtacta aatggtagat ttaagaactt
      181 taggatattc acagcagcaa caggaaaaga tcaagcccaa agttaggtcg acagtcgcgc
      241 agcatcacga agcgctggtt ggtcatgggt ttacacatgc ccacatcgta gccttatcgc
      301 agcaccctgc agcccttggc acggtcgccg tcaagtacca ggacatgatt gcggcgttgc
      361 cggaagccac acatgaggcg atcgtcggtg tggggaaaca gtggagcgga gcccgagcgc
      421 ttgaggccct gttgacggtc gcgggagagc tgagagggcc tccccttcag ctggacacgg
      481 gccagttgct gaagatcgcg aagcggggag gagtcacggc ggtcgaggcg gtgcacgcgt
      541 ggcgcaatgc gctcacggga gcacccctca atctgacccc ggatcaggtg gttgcgatcg
      601 ccagtaatgg cggaggcaaa caagcactcg aaactgtaca gcgcctcctg ccggtactgt
      661 gccaagatca tggcttgacg cctgagcaag tcgtagctat tgcatcaaac atcggtggta
      721 aacaggcgct ggaaaccgta caacgattac tccctgtctt atgccaggca cacggtctga
      781 cccccgacca agttgtagcc attgcgtcta atggcggcgg gaaacaggcc ctggaaacgg
      841 tccaacgtct gttacccgtt ctgtgtcagg ctcacggtct gacccctgcc caggtagttg
      901 caattgccag caacatcggc gggaaacaag cgctggagac tgtgcagcgt ctgctccctg
      961 tgttatgcca agatcatggg ctcactccgg atcaggtggt ggccatcgct tccaatattg
     1021 gcggtaaaca ggcgctggag acagtgcaac gacttttacc tgttctctgc caggatcatg
     1081 gtctaactcc cgagcaggtc gtcgccatcg cctctcatga cggcgggaaa caagcgttgg
     1141 aaactgtcca gcgactcctg ccggttttgt gccaggccca cgggcttact cctgaccagg
     1201 tagttgcgat cgcgtcaaat gggggtggca aacaagccct cgaaaccgtg caacgcctgc
     1261 tgcccgtctt gtgccaagct catgggctga ctccggcgca agtagtcgcg attgcgagcc
     1321 acgatggcgg taagcaggca ctggaaacgg ttcagcgcct gctcccggtt ctatgccagg
     1381 atcacggcct gaccccggac caggtcgtcg cgatcgcgtc aaatatcggt ggcaaacaag
     1441 ctttggagac agtacagcgc ctgttaccag tgctttgcca ggaccatggt ctgacccctg
     1501 agcaagtagt ggcgatcgct tctaatattg ggggcaaaca agcgctggaa acagtacagc
     1561 gtctgttacc ggtcctatgc caggcacatg gcctgacccc tgatcaggtg gtagccattg
     1621 ccagtcatga tggcggtaaa caggcgcttg agactgtcca acgtctgctg ccggtcctct
     1681 gtcaggctca tggcctgacg ccagctcaag tcgtggctat cgcttcgcat gatggcggaa
     1741 aacaggcact ggagactgtg cagcgactgt tgccagttct gtgtcaggat cacggtttaa
     1801 ctccggacca ggtggtcgct attgcgtcga atgggggcgg taaacaagcg ctggaaactg
     1861 tgcaacgttt gctcccagtt ctgtgccagg accatgggct gactccggaa caggtagtgg
     1921 ccattgcttc taatattggt gggaaacagg cgctggaaac cgtgcagcgc ctgcttccag
     1981 tgctttgcca ggcccatggc ctgacgccag atcaggtggt tgctatagcc agcaatggcg
     2041 gcggtaaaca ggccctcgaa accgtccagc gcctgctccc tgtgctgtgc caggcccatg
     2101 ggcttacccc agcgcaagta gtggcgattg cgtctaatat tggtggtaaa caggcgttgg
     2161 agactgtaca acgcctgctg ccagttttat gccaagatca tggtctgacc cctgagcagg
     2221 tagtggctat tgcatccaac aacgggggca gacccgcact ggagtcaatc gtggcccagc
     2281 tttcgaggcc ggaccccgcg ctggccgcac tcactaatga tcatcttgta gcgctggcct
     2341 gcctcggcgg acgacccgcc ttggatgcgg tgaagaaggg gctcccgcac gcgcctgcat
     2401 tgattaagcg gaccaacaga aggattcccg agaggacatc acatcgagtg gcagatcacg
     2461 cgcaagtggt ccgcgtgctc ggattcttcc agtgtcactc ccaccccgca caagcgttcg
     2521 atgacgccat gactcaattt ggtatgtcga gacacggact gctgcagctc tttcgtagag
     2581 tcggtgtcac agaactcgag gcccgctcgg gcacactgcc tcccgcctcc cagcggtggg
     2641 acaggattct ccaagcgagc ggtatgaaac gcgcgaagcc ttcacctacg tcaactcaga
     2701 cacctgacca ggcgagcctt catgcgttcg cagactcgct ggagagggat ttggacgcgc
     2761 cctcgcccat gcatgaaggg gaccaaactc gcgcgtcata ataggtccag gcatcaaata
     2821 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     2881 gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatatacta
     2941 gtagcggccg ctgcagtccg gcaaaaaagg gcaaggtgtc accaccctgc cctttttctt
     3001 taaaaccgaa aagattactt cgcgttatgc aggcttcctc gctcactgac tcgctgcgct
     3061 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaatc tcgagtcccg
     3121 tcaagtcagc gtaatgctct gccagtgtta caaccaatta accaattctg attatttgcc
     3181 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgtgcg
     3241 cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg
     3301 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
     3361 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
     3421 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
     3481 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
     3541 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
     3601 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
     3661 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
     3721 ctcgctctct acaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
     3781 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
     3841 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
     3901 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct
     3961 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
     4021 atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat
     4081 aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca acgcagaccg
     4141 ttccgtggca aagcaaaagt tcaaaatcac caactggtcc acctacaaca aagctctcat
     4201 caaccgtggc tccctcactt tctggctgga tgatggggcg attcaggcct ggtatgagtc
     4261 agcaacacct tcttcacgag gcagacctca gcgctagcgg agtgtatact ggcttactat
     4321 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     4381 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     4441 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     4501 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     4561 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     4621 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     4681 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     4741 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     4801 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4861 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     4921 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     4981 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     5041 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     5101 caagaagatc atcttattaa ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5161 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5221 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5281 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5341 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5401 ataccgcgat acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5461 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattccatgg
     5521 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5581 tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctggt
     5641 cggccaattc gcgctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5701 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5761 gtattgggcg ccagggtggt ttttcttttc accagtgaga ctggcaacag ctgattgccc
     5821 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     5881 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctatc ttcggtatcg
     5941 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     6001 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     6061 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     6121 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     6181 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     6241 agatgctcca cgcccagtcg cgtaccgtcc tcatgggaga aaataatact gttgatgggt
     6301 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     6361 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     6421 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     6481 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     6541 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     6601 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     6661 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     6721 ccggcatact ctgcgacatc gtataacgtt actggtttca tattcaccac cctgaattga
     6781 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggcgcgc
     6841 cgcttcgtca ggccacatag ctttcttgtt ctgatcggaa cgatcgttgg ctgtgtt
//
