LOCUS       sequence-353272-        7035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of TRX-AviTag-9S-S2 [rpoA].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7035)
  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 7035)
  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-avi-9S-S2 [rpoA]
FEATURES             Location/Qualifiers
     source          1..7035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      62..517
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(149..168)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     359..380
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        543..647
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(648..716,717..1508)
                     /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(866..885)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1679..2267
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2168..2187
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2421..2438
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2453..2592
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2518..2537)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2678..2700
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2694..2885)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3694..4776)
                     /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     4738..4757
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4777..4854)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5060..5079
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5167..5186
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5167..5185
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5186..5210
                     /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             5225..5247
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5255..5581
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             5603..5620
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5630..5650
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5657..5701
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6929..6946
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6999..7017)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      join(7013..7035,1..25)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
       61 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      121 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      181 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      241 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      301 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      361 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      421 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      481 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
      541 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      601 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      661 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
      721 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
      781 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
      841 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
      901 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
      961 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1021 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1081 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1141 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     1201 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1261 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1321 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1381 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1441 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1501 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1561 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1621 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1681 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1741 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1801 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     1861 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1921 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1981 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2041 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2101 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2161 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2221 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2281 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2341 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2401 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     2461 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     2521 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     2581 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2641 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2701 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     2761 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     2821 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     2881 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     2941 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     3001 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     3061 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     3121 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     3181 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     3241 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     3301 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc
     3361 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
     3421 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     3481 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     3541 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
     3601 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     3661 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3721 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3781 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     3841 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     3901 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     3961 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     4021 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     4081 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     4141 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     4201 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     4261 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     4321 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     4381 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     4441 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     4501 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     4561 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     4621 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     4681 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     4741 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     4801 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     4861 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     4921 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     4981 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     5041 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     5101 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg
     5161 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
     5221 taattttgtt taactttaag aaggagatat acatatgagc gataaaatta ttcacctgac
     5281 tgacgacagt tttgacacgg atgtactcaa agcggacggg gcgatcctcg tcgatttctg
     5341 ggcagagtgg tgcggtccgt gcaaaatgat cgccccgatt ctggatgaaa tcgctgacga
     5401 atatcagggc aaactgaccg ttgcaaaact gaacatcgat caaaaccctg gcactgcgcc
     5461 gaaatatggc atccgtggta tcccgactct gctgctgttc aaaaacggtg aagtggcggc
     5521 aaccaaagtg ggtgcactgt ctaaaggtca gttgaaagag ttcctcgacg ctaacctggc
     5581 cggttctggt tctggccata tgcaccatca tcatcatcat tcttctggtg agaatcttta
     5641 ttttcagggc gggtccgggt tgaatgatat tttcgaagca cagaaaattg aatggcatga
     5701 ggccatggga aattctgttg tttcttggaa ctctatgatt actgcttatg ctagaaacgg
     5761 tagaatcgaa gaggctagaa aattgtttga tgaaatgcct gagagagatg ttgtttcttg
     5821 gaatgctatg atttctggat acgctaagaa cggtgaactt gatgaggcta gaaaattgtt
     5881 cgatgaaatg ccagaaaggg atgttgtttc ttggacagct atgatttctg gatatgctca
     5941 aaatggtaga atcgatgaag ctagagagct ttttgataag atgcctgaga gaaatgttgt
     6001 ttcttggaac gcaatgattt ctggttacgc tcagaacggt aggattgatg aagctagaga
     6061 gttgttcgat aaaatgccag aaagatcagt tgtttcttgg accgcaatga tttctggata
     6121 cgcacagaat ggtaggatag atgaggctag agagcttttc gataaaatgc cagagaggaa
     6181 cgttgtttct tggaacgcta tgatttctgg ttatgctcag aacggtagga tagacgaggc
     6241 tagagagttg tttgataaga tgcctgaaag atcagttgtt tcttggacgg caatgatttc
     6301 tggttatgca cagaatggta ggatcgacga ggctagagag ctttttgaca agatgccaga
     6361 acgtgacgtt gtttcttgga atgcaatgat ttctggatat gcacagaacg gtaggataga
     6421 tgaagctaga gagcttttcg acaagatgcc agaaagagac gttgtttctt ggactgctat
     6481 gatttctgct tatgctcaat gtggacattt ggaagaggct agaaaaatct ttgataggat
     6541 gccagaacgt gatgttgttt cttggaacgc gatgattgct ggatacgctc aaaacggaca
     6601 tggtgaagag gctcttgaat tgtttagaga gatgcagctt gaaggtatca aaccatctga
     6661 ggttactttc acatctgttc tttctgcttg ttctcatgct ggattggttg aagagggttg
     6721 ggaatatttc gtttctatgg cttctgatta cggaattgag cctagaagag aacattatgg
     6781 ttgtatggtt gatcttttgg gaagagctgg tagacttgaa gaggctgaag agttgatcga
     6841 gtctatgcct tttgaaccaa atgctgttgt ttggagaact ttccttcagg gttgtaagac
     6901 atacggtatg ttggattaat aactcgagca ccaccaccac caccactgag atccggctgc
     6961 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     7021 accccttggg gcctc
//
