LOCUS       sequence-353268-        7506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of TRX-9S-DYW [rpoA].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7506)
  AUTHORS   Bernath-Levin K, Schmidberger J, Honkanen S, Gutmann B, Sun YK, 
            Pullakhandam A, Colas des Francs-Small C, Bond CS, Small I
  TITLE     Cofactor-independent RNA editing by a synthetic S-type PPR protein.
  JOURNAL   Synth Biol (Oxf). 2021 Dec 23;7(1):ysab034. doi: 
            10.1093/synbio/ysab034. eCollection 2021.
  PUBMED    35128071
REFERENCE   2  (bases 1 to 7506)
  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: "Synth Biol 
            (Oxf)."; date: "2021-12-23"; volume: "7(1):ysab034. doi"; pages: " 
            10.1093/synbio/ysab034. eCollection 2021"
COMMENT     Sequence Label: pETM20-9S-DYW [rpoA]
FEATURES             Location/Qualifiers
     source          1..7506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..42
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     130..149
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        130..148
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    149..173
                     /label=lac operator
                     /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)."
     RBS             188..210
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             218..544
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             566..583
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             593..613
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2447..2494
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2531..2986
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2618..2637)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2828..2849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3012..3116
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3117..3185,3186..3977)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3335..3354)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4148..4736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4637..4656
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4890..4907
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4922..5061
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4987..5006)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5147..5169
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5163..5354)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6163..7245)
                     /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     7207..7226
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7246..7323)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
       61 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc
      121 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
      181 aaataatttt gtttaacttt aagaaggaga tatacatatg agcgataaaa ttattcacct
      241 gactgacgac agttttgaca cggatgtact caaagcggac ggggcgatcc tcgtcgattt
      301 ctgggcagag tggtgcggtc cgtgcaaaat gatcgccccg attctggatg aaatcgctga
      361 cgaatatcag ggcaaactga ccgttgcaaa actgaacatc gatcaaaacc ctggcactgc
      421 gccgaaatat ggcatccgtg gtatcccgac tctgctgctg ttcaaaaacg gtgaagtggc
      481 ggcaaccaaa gtgggtgcac tgtctaaagg tcagttgaaa gagttcctcg acgctaacct
      541 ggccggttct ggttctggcc atatgcacca tcatcatcat cattcttctg gtgagaatct
      601 ttattttcag ggcgccatgg gaaattctgt tgtttcttgg aactctatga tcactgctta
      661 tgctagaaac ggtagaattg aagaggctag aaagttgttt gatgagatgc ctgaaagaga
      721 tgttgtttct tggaatgcta tgatctctgg atacgctaaa aacggtgagc ttgatgaagc
      781 tagaaagttg ttcgatgaga tgccagagag ggatgttgtt tcttggacag ctatgatctc
      841 tggatatgct caaaatggta gaattgatga ggctagagaa cttttcgata agatgcctga
      901 aagaaacgtt gtttcttgga acgcaatgat ctctggttac gctcagaacg gtaggatcga
      961 tgaggctaga gaattgttcg ataagatgcc agagagatca gttgtttctt ggaccgcaat
     1021 gatctctgga tacgcacaga atggtaggat agatgaagct agagaacttt ttgataaaat
     1081 gccagagagg aatgttgttt cttggaatgc gatgatctct ggttatgcac agaacggtag
     1141 gatagacgaa gctagagaat tgtttgataa gatgcctgaa agatcagttg tttcttggac
     1201 ggcaatgatc tctggttatg ctcagaatgg taggattgac gaagctagag aacttttcga
     1261 caaaatgcca gagcgtgacg ttgtttcttg gaatgcaatg atctctggat atgcacagaa
     1321 cggtagaatc gacgaagcta gagaactttt cgacaagatg ccagagcgtg atgttgtttc
     1381 ttggactgct atgatctctg cttatgctca atgtggacat ttggaagagg ctagaaagat
     1441 ttttgataga atgccagaga gagacgttgt ttcttggaat gcaatgatct ctggttacgc
     1501 tatgcatgga catggtaaag aggctcttga attgtttgaa gagatgcaac agtctggtat
     1561 taagccttct catgttactt tcacaggagt tctttctgct tgttctcatg ctggtttggt
     1621 tgatgaggga agacaatact tcaactctat gaagaaggat tacggtatcg aaccaagagt
     1681 tgagcattac ggatgtatgg ttgatctttt gggaagagct ggtagacttg atgaggctta
     1741 tgagtttatt gagtctatgc ctattgaacc aaatgctgtt gtttggggtg ctcttttggg
     1801 tgcttgtagg attcatggaa acgttgaatt gggagagaga gctgctgaga aactttttga
     1861 attggagcca gaatcttctg gtaactacgt tcttttgtct aacatctatg cttctgctgg
     1921 aagatgggat gatgttgcta aggttagaaa aatgatgaag gaaagaggta tcaagaaaga
     1981 gcctggatgt tcttggattg aagttaagaa taaggttcat gagttcgttg ctggagatag
     2041 atcacatcca caatctgaag aaatctatgc taaacttgaa gagttgtctg aaaagatgaa
     2101 agaggctgga tatgttcctg atacttcttt tgttcttcat gatgttgaag aggaagagaa
     2161 agaacagatg ttgtcttacc attctgagaa acttgctatc gctttcggtt tgatttctac
     2221 tcctccagga acaccaatca gaatcgttaa aaatcttaga gtttgtggag attgtcatac
     2281 agctattaaa ttcatctcta agattgttgg aagagaaatt atcgttagag attctaacag
     2341 attccatcat ttcaaggatg gttcttgttc ttgtggagat tattggtaac tcgagaaata
     2401 gaaggaacat gtattacacg tgcaaaatct gagagctgag caataactag cataacccct
     2461 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     2521 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2581 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2641 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2701 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2761 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2821 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2881 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2941 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     3001 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3061 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3121 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3181 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3241 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3301 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3361 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3421 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3481 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3541 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3601 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3661 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3721 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3781 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3841 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3901 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3961 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4021 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4081 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4141 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4201 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4261 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4321 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4381 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4441 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4501 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4561 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4621 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4681 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4741 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4801 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4861 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4921 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
     4981 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     5041 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5101 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5161 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     5221 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     5281 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     5341 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     5401 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     5461 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     5521 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     5581 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     5641 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     5701 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     5761 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     5821 ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca
     5881 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc
     5941 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc
     6001 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     6061 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     6121 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     6181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6241 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     6301 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     6361 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     6421 atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc
     6481 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     6541 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     6601 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     6661 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     6721 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     6781 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     6841 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     6901 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     6961 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     7021 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     7081 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     7141 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     7201 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     7261 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     7321 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     7381 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     7441 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     7501 cgaagt
//
