LOCUS       sequence-194851-        6668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows expression of a modified version of the fission yeast NatA 
            complex - chloramphenicol marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6668)
  AUTHORS   Eastwood T, Baker K, Brooker H, Frank S, Mulvihill DP
  TITLE     An enhanced recombinant amino-terminal acetylation system and novel 
            in vivo high-throughput screen for molecules affecting 
            alpha-synuclein oligomerisation.
  JOURNAL   FEBS Lett. 2017 Feb 18. doi: 10.1002/1873-3468.12597.
  PUBMED    28214355
REFERENCE   2  (bases 1 to 6668)
  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: "FEBS Lett. 
            2017 Feb 18. doi: 10.1002/1873-3468.12597."
COMMENT     Sequence Label: pNatA (pACYCduet-naa10-naa15)
FEATURES             Location/Qualifiers
     source          1..6668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        2380..2398
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2399..2423
                     /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)."
     CDS             3024..3068
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3106..3124)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3120..3167
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3388..4047)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3962..3981
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4048..4150)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4542..4559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4676..5221)
                     /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(4756..4775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5446..6528)
                     /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     6490..6509
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6529..6606)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(6652..6668,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
      121 ctcgatggcc aaagttcaac tgagcccgaa ggagattacc ctttttagaa cagcattaaa
      181 atgctatgaa acaaaacagt acaaaaaagg attaaaggct atcgaacccc tgttagaaag
      241 gcatccagag catggagaat ccttagcaat taagggaatt ttattacact ctttaggtaa
      301 taccaaagaa ggatacgaca acgtacgact tggtcttcgc aacgatgttg gatccggtgt
      361 ttgttggcac attttcggac taattagcag agccgataaa gattatgtac aggccgctaa
      421 atgttacatt aatgctcata agcttgagaa aaacaactcg agtcttttgc gtgacttggc
      481 gttactccaa tctcaacttc gtcaatacaa ggctctagca gacactagga atgctttgct
      541 gcaggacaat ccaggagtac gtgccaattg gagcgcttta gcagtagctc aatttcttcg
      601 aggggaatac gctagtgcat ataaaatagt ggatgcattt gaatccacta taaatcaagg
      661 tgtgcccgtt gatacccaag aagaatccga agccatgtta ttcatgaacc ttgttatatt
      721 aaaaaaagat ggtgttgagg atgcctataa gcatttactg tcaatagaaa agaaggtatt
      781 ggatcgggtt gcgtttttgg agactcgggc agagtacgag ctttatcttt ccaaaatgga
      841 agaagccaaa tcaaccatat atcttctgct tgatcgaaat ccagacaatc accaatatta
      901 ttataatctt caaagagcgt atgggtatga ggatgccagt ggtaaagtat tggatagtgc
      961 tgagtggttg aatttatatt ctcaacttgc taaaaggtat cccaaatctg agtgccctac
     1021 gagactacct ttagaaaagc ttgaagggga tgagttttta actcatgtag atctttactt
     1081 gcgcaaaaag cttaagcgtg gaatcccatc tgtgttcgtc gatgtcaaat ctttgtacaa
     1141 agataccaaa aaatgcaaag ttgttgagga tttggttagt aaatacgctt cctctctttc
     1201 tacgaccaac aaattttctg aagatgacga taattcacag attgaaattc ccaccacgct
     1261 tttatggaca tattactttt tagctcagca ttttgatcat gttggtgagt tggaaaaagc
     1321 tgagaaatat gttgatttgg caattgatca taccccaacg cttgtggagc tgttcatgac
     1381 gaaagctagg atttccaaac ataagggaga gcttcaaact gccatggaga ttatggatca
     1441 tgctcgtaaa cttgatctgc aagatagatt tattaacggc aagtgtgcta aatatatgct
     1501 ccgtaatgat gaaaatgaac ttgccgccaa aacggtatcg ttgtttacac gcaatgaggc
     1561 tgttggtggt gctgttggcg atcttgcaga tatgcaatgc ctgtggtaca tgcttgaaga
     1621 tgggaaatca tttgctagac aaaaaaaatt tgctttggct ttgaaacgtt ttagtactgt
     1681 ttttaaaata tttgatactt gggctgatga tcaatttgat ttccacttct tcgcatttcg
     1741 caagggatct cttcgtacat acctggactt aatgtcttgg gaagacagtg tttatgacga
     1801 tccttcgttt agagaggctg cccaagggtc tattgaaatc tattttgcct tatttgattt
     1861 accatttgct aaatatagcc ctaaattacc cgattttgag aagctttcta gtggagaaat
     1921 aaatgaagaa gaggaaaaga agatatacaa gaagctgaag aaggacctta gtaaacgatt
     1981 ggaacgtgct gagaagctaa aagaggccga taaatctcgt gcaaaatctg aggacggtat
     2041 gccggtaaag tatgatgagg atcctcttgg ggagaactta gttgctacta gtgaaccttt
     2101 gaaagaagct caaaaatgtc ttgaaaaatt actcccctat ggagataaaa acccaagtgc
     2161 ttacattttg gcggctcaac tttacactcg tttgaaaaat ttcgataccg ctagtaaata
     2221 cctcgaacaa gctaaagtaa ttttgggaca gaatgaccca actgtgatat ctacagagaa
     2281 gttttataac tctatcaaaa ctcagtctaa ctaagcggcc gcataatgct taagtcgaac
     2341 agaaagtaat cgtattgtac acggccgcat aatcgaaatt aatacgactc actatagggg
     2401 aattgtgagc ggataacaat tccccatctt agtatattag ttaagtataa gaaggagata
     2461 tacatatgga tattcgtccc gcgcgcatta gtgacctgac tggtatgcag aattgcaacc
     2521 tgcacaatct gcccgaaaat taccaactga aatattatct ttatcacgcg atcagctggc
     2581 ctatgctgag ctatgttgca actgacccga aaggtcgtgt tgttggctat gtccttgcaa
     2641 aaatggaaga agagccaaag gatggtattc cgcatggcca tatcactagc gtttccgtga
     2701 tgcgtagtta ccgccattta ggtctcgcca aacgcctgat ggttcaaagc cagcgcgcga
     2761 tggtcgaagt gtatggcgcc aaatatatgt ccctgcatgt gcggaagagc aatcgtgccg
     2821 cgatccattt ataccgtgat accctgcagt ttgacgttca gggcattgaa tccaagtatt
     2881 atgccgacgg ggaggacgcg tatgcgatgc ataaagactt ttcaacgctg aaattcgata
     2941 ctccggagac aaatgatgaa ctggcaaaga cggtacaaag tttagcattg aacaattgaa
     3001 gatctggtac cctcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca
     3061 tggactcgtc tactagcgca gcttaattaa cctaggctgc tgccaccgct gagcaataac
     3121 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg
     3181 catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat
     3241 agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt
     3301 cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcaac caggcgttta
     3361 agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagtactg
     3421 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     3481 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tagtgaaaac
     3541 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     3601 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     3661 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     3721 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     3781 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga actccggatg
     3841 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     3901 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     3961 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     4021 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
     4081 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     4141 gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg
     4201 gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcggcgca
     4261 aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt atgatggtgt ttttgaggtg
     4321 ctccagtggc ttctgtttct atcagctgtc cctcctgttc agctactgac ggggtggtgc
     4381 gtaacggcaa aagcaccgcc ggacatcagc gctagcggag tgtatactgg cttactatgt
     4441 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg
     4501 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
     4561 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
     4621 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
     4681 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
     4741 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
     4801 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
     4861 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
     4921 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4981 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
     5041 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
     5101 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
     5161 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
     5221 agaagatcat cttattaatc agataaaata tttctagatt tcagtgcaat ttatctcttc
     5281 aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg
     5341 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     5401 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     5461 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5521 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     5581 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     5641 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     5701 ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat
     5761 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     5821 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     5881 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     5941 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     6001 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     6061 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     6121 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     6181 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     6241 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     6301 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     6361 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     6421 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     6481 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     6541 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     6601 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa ttaatacgac
     6661 tcactata
//
