LOCUS       sequence-212221-        6897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PTac driving TALEsp2.
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_40
FEATURES             Location/Qualifiers
     source          1..6897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(3..8,9..24,25..31)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..56
                     /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        74..123
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     terminator      2819..2890
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2906..2933
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2940..2960
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2961..3018
                     /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(3176..3967)
                     /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(4562..5107)
                     /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(4642..4661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5575..5593
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5597..5640)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(5683..6765)
                     /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     6727..6746
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcgctc acaattctat
       61 ggactatgtt tagctgtcac cggatgtgct ttccggtctg atgagtccgt gaggacgaaa
      121 cagcctctac aaataatttt gtttaatcac acaggaaagt actaaatggt agatttaaga
      181 actttaggat attcacagca gcaacaggaa aagatcaagc ccaaagttag gtcgacagtc
      241 gcgcagcatc acgaagcgct ggttggtcat gggtttacac atgcccacat cgtagcctta
      301 tcgcagcacc ctgcagccct tggcacggtc gccgtcaagt accaggacat gattgcggcg
      361 ttgccggaag ccacacatga ggcgatcgtc ggtgtgggga aacagtggag cggagcccga
      421 gcgcttgagg ccctgttgac ggtcgcggga gagctgagag ggcctcccct tcagctggac
      481 acgggccagt tgctgaagat cgcgaagcgg ggaggagtca cggcggtcga ggcggtgcac
      541 gcgtggcgca atgcgctcac gggagcaccc ctcaacctga ccccggacca ggtagtcgcg
      601 attgcttcac atgacggggg taaacaagcg ctggaaacgg tgcagcgtct gctaccggtg
      661 ttatgtcagg atcatgggct cacgccggaa caggtagtgg caattgcgag tcatgacggt
      721 ggcaaacagg ccctggaaac cgtacagcgg ctgctaccgg tgctgtgtca agcgcatggc
      781 ctgactccgg accaagttgt agccattgcc tcgaacgggg gcggcaagca ggcgctggag
      841 actgttcaac gtctgctccc cgttctgtgt caggcgcatg gcctgacgcc tgcgcaggtc
      901 gtggcgatcg cttcaaacat cggtgggaag caagccctgg agactgtcca aagactgttg
      961 ccagtgttgt gtcaagatca tggcttaacc ccagatcagg tggttgcgat tgcatcaaat
     1021 ggaggtggta aacaggcgct ggagactgtg cagcgcctgt tgccggttct gtgccaagat
     1081 catgggctga ctccggaaca ggttgtggct atcgcaagca atattggtgg caagcaggcc
     1141 ctggaaacag tacagcgcct gctgcctgta ttgtgtcaag cccacggtct tacccccgat
     1201 caggtagtcg ccatagcatc gcacgacggc gggaagcagg cccttgagac tgtacaacgc
     1261 ctcctgccgg ttttgtgcca ggcgcacggc ctgacgccag cccaggtggt tgcgatagcc
     1321 agtaataacg gcggtaagca agcccttgaa accgttcaac gtttgctgcc agtgctgtgc
     1381 caggatcacg gcctgacccc ggatcaggta gtcgccattg ctagcaacat tggtggcaaa
     1441 caagcactgg agacagttca acgcttactg cccgtgcttt gccaggatca tggactgacc
     1501 ccagagcaag tggtcgcgat tgcctcgcat gacggaggta aacaggccct tgagactgtc
     1561 cagcgtctgc tgccggtcct ttgccaggct catgggctga cgcccgacca ggtggtagca
     1621 atcgcttcga acggcggagg caaacaggcg ttagaaaccg ttcaacgtct gttaccggtg
     1681 ctgtgccaag ctcatggctt aaccccggcc caggtagtcg caatcgctag tcatgatggt
     1741 gggaaacagg cattagaaac agtacaacgt ctgctcccgg tcctctgtca ggatcacggt
     1801 ctgaccccgg atcaggttgt agcaatcgct agcaatattg gtggtaaaca ggcgctggaa
     1861 acagtgcaaa gattattacc agttctgtgt caggaccatg gcctgacacc tgagcaggtc
     1921 gtagcgatcg caagtaatat tggaggtaaa caggccctgg aaaccgtgca gcgtctgctg
     1981 cctgtgctct gtcaagcgca tggtctgact ccggatcagg ttgtcgcgat cgccagtaac
     2041 gggggaggga aacaggcact cgagactgtg cagagattgc tgccggtcct gtgtcaggcg
     2101 catggtctga ccccagcgca ggtcgtggca atcgcttcca acataggtgg taaacaggcc
     2161 ctcgaaactg tccaacggct gttaccggta ctgtgccagg atcatggtct gacccctgag
     2221 caggtagtgg ctattgcatc caacggaggg ggcagacccg cactggagtc aatcgtggcc
     2281 cagctttcga ggccggaccc cgcgctggcc gcactcacta atgatcatct tgtagcgctg
     2341 gcctgcctcg gcggacgacc cgccttggat gcggtgaaga aggggctccc gcacgcgcct
     2401 gcattgatta agcggaccaa cagaaggatt cccgagagga catcacatcg agtggcagat
     2461 cacgcgcaag tggtccgcgt gctcggattc ttccagtgtc actcccaccc cgcacaagcg
     2521 ttcgatgacg ccatgactca atttggtatg tcgagacacg gactgctgca gctctttcgt
     2581 agagtcggtg tcacagaact cgaggcccgc tcgggcacac tgcctcccgc ctcccagcgg
     2641 tgggacagga ttctccaagc gagcggtatg aaacgcgcga agccttcacc tacgtcaact
     2701 cagacacctg accaggcgag ccttcatgcg ttcgcagact cgctggagag ggatttggac
     2761 gcgccctcgc ccatgcatga aggggaccaa actcgcgcgt cataataggt ccaggcatca
     2821 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     2881 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttatat
     2941 actagtagcg gccgctgcag tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt
     3001 tctttaaaac cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac tgactcgctg
     3061 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatctcgagt
     3121 cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattatt
     3181 tgccgactac cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg
     3241 tgcgcgaggc caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga
     3301 cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg
     3361 cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct
     3421 catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa
     3481 atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa
     3541 cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct
     3601 cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag
     3661 ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga
     3721 gaatctcgct ctctacaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc
     3781 gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct
     3841 tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat
     3901 ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt
     3961 ccctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     4021 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt
     4081 gaataaatcg aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag
     4141 accgttccgt ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc
     4201 tcatcaaccg tggctccctc actttctggc tggatgatgg ggcgattcag gcctggtatg
     4261 agtcagcaac accttcttca cgaggcagac ctcagcgcta gcggagtgta tactggctta
     4321 ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct
     4381 gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac
     4441 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat
     4501 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt
     4561 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt
     4621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc
     4681 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt
     4741 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg
     4801 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4861 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
     4921 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
     4981 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
     5041 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
     5101 atctcaagaa gatcatctta ttaaggggtc tgacgctcag tggaacgaaa actcacgtta
     5161 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5221 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5281 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5341 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5401 aatgataccg cgatacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5461 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattcc
     5521 atggtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     5581 cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa ggatgatttc
     5641 tggtcggcca attcgcgcta acttacatta attgcgttgc gctcactgcc cgctttccag
     5701 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5761 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagactggca acagctgatt
     5821 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     5881 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tatcttcggt
     5941 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     6001 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     6061 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     6121 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     6181 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     6241 gaccagatgc tccacgccca gtcgcgtacc gtcctcatgg gagaaaataa tactgttgat
     6301 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     6361 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     6421 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     6481 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     6541 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     6601 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     6661 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     6721 gacaccggca tactctgcga catcgtataa cgttactggt ttcatattca ccaccctgaa
     6781 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggc
     6841 gcgccgcttc gtcaggccac atagctttct tgttctgatc ggaacgatcg ttggctg
//
