LOCUS       sequence-353266-        7104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of TRX-9P-S2 [rpoA].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7104)
  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 7104)
  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: pETM20-9P-S2 [rpoA]
FEATURES             Location/Qualifiers
     source          1..7104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..37)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      33..80
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      117..572
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(204..223)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     414..435
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        598..702
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(703..771,772..1563)
                     /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(921..940)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1734..2322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2223..2242
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2476..2493
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2508..2647
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2573..2592)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2733..2755
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2749..2940)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3749..4831)
                     /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     4793..4812
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4832..4909)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5115..5134
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5222..5241
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5222..5240
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5241..5265
                     /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             5280..5302
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5310..5636
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             5658..5675
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5685..5705
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             7053..7070
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
       61 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
      121 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      181 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      241 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      301 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
      361 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
      421 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
      481 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
      541 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
      601 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      661 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
      721 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
      781 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
      841 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
      901 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
      961 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1021 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1081 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1141 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     1201 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     1261 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1321 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1381 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1441 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     1501 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1561 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     1621 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     1681 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1741 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1801 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1861 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     1921 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1981 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2041 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2101 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2161 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2221 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2281 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2341 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2401 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2461 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     2521 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     2581 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     2641 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     2701 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     2761 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     2821 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     2881 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     2941 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     3001 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     3061 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     3121 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     3181 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     3241 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     3301 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     3361 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc
     3421 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     3481 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     3541 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     3601 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     3661 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     3721 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3781 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3841 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     3901 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     3961 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     4021 ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     4081 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     4141 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     4201 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     4261 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     4321 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     4381 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     4441 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     4501 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     4561 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     4621 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     4681 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     4741 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     4801 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     4861 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     4921 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     4981 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     5041 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     5101 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     5161 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tcgatctcga tcccgcgaaa
     5221 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt
     5281 ttgtttaact ttaagaagga gatatacata tgagcgataa aattattcac ctgactgacg
     5341 acagttttga cacggatgta ctcaaagcgg acggggcgat cctcgtcgat ttctgggcag
     5401 agtggtgcgg tccgtgcaaa atgatcgccc cgattctgga tgaaatcgct gacgaatatc
     5461 agggcaaact gaccgttgca aaactgaaca tcgatcaaaa ccctggcact gcgccgaaat
     5521 atggcatccg tggtatcccg actctgctgc tgttcaaaaa cggtgaagtg gcggcaacca
     5581 aagtgggtgc actgtctaaa ggtcagttga aagagttcct cgacgctaac ctggccggtt
     5641 ctggttctgg ccatatgcac catcatcatc atcattcttc tggtgagaat ctttattttc
     5701 agggcgccat gggaaatgat gtcgtcacgt acaacaccct tatcgacgga cttgctaagg
     5761 ctggtcgtct ggaagaagct cttcagctgt ttcaagagat gaaggaaaag ggcgtaaagc
     5821 ctgacgtggt aacgtacaat acgcttatcg atggtctggc gaaggctggc cgtcttgaag
     5881 aggccctgca gctgtttcag gaaatgaaag aaaagggcgt taaacctgat gtagtaacct
     5941 acacgacatt gatcgacgga cttgctaagg ccggacgctt agaggaggcc cttcaactgt
     6001 tccaggagat gaaggagaaa ggagtaaaac ctaacgtggt cacttataat actttgattg
     6061 acggtttggc aaaggcagga cgtttggaag aggccttaca attgtttcaa gaaatgaaag
     6121 agaagggcgt gaaaccgtct gttgtaacct atactacctt aatcgatggg ctggcaaaag
     6181 cgggtcgttt agaggaagct ttacagttgt tccaggagat gaaagagaaa ggtgtcaagc
     6241 ccaacgttgt cacgtacaac acccttattg atggtctggc caaagccggt cgcttagagg
     6301 aagccttgca gcttttccag gagatgaaag agaagggtgt taagccaagc gtggttacat
     6361 acaccacttt aatcgatgga ttagcaaagg caggacgcct ggaggaggcc ctgcaacttt
     6421 ttcaagagat gaaggagaaa ggggtcaaac ctgatgtggt tacatacaat accttgatcg
     6481 acggacttgc taaggcgggt cgccttgaag aagcgttaca gttgttccaa gaaatgaaag
     6541 aaaagggagt gaaaccggat gtggtgactt acacgacgct tatcgacggg cttgctaagg
     6601 ccgggcgttt agaagaagct ttgcaactgt ttcaagaaat gaaggaaaaa ggggttaagc
     6661 cagatgttgt ttcttggaac gcgatgattg ctggatacgc tcaaaacgga catggtgaag
     6721 aggctcttga attgtttaga gagatgcagc ttgaaggtat caaaccatct gaggttactt
     6781 tcacatctgt tctttctgct tgttctcatg ctggattggt tgaagagggt tgggaatatt
     6841 tcgtttctat ggcttctgat tacggaattg agcctagaag agaacattat ggttgtatgg
     6901 ttgatctttt gggaagagct ggtagacttg aagaggctga agagttgatc gagtctatgc
     6961 cttttgaacc aaatgctgtt gtttggagaa ctttccttca gggttgtaag acatacggta
     7021 tgttggatta aggtaccgcg gccgcactcg agcaccacca ccaccaccac tgagatccgg
     7081 ctgctaacaa agcccgaaag gaag
//
