LOCUS       sequence-486030-        9048 bp    DNA     circular SYN 19-SEP-2025
DEFINITION  Recombineering Plasmid 9. Positive control to test for successful 
            editing using AraC S170 LOV (silent mutation of P190, 90bp retron).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9048)
  TITLE     Phage-Assisted Evolution of Allosteric Protein Switches
REFERENCE   2  (bases 1 to 9048)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 19, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: RcP9_AraC_S170_LOV_P190_Sil_90bp
FEATURES             Location/Qualifiers
     source          1..9048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(888..1970)
                     /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     1932..1951
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1971..2048)
                     /gene="lacI"
                     /label=lacI promoter
     CDS             complement(2153..3427)
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKPHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     protein_bind    3497..3515
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      complement(3679..3706)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3798..3884)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4037..4054
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     protein_bind    4125..4141
                     /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)."
     terminator      8199..8230
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(8246..8905)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAKCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     8820..8839
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ctgcggcggg cgctgcggac acatacaaag ttacccacag attccgtgga taagcagggg
       61 actaacatgt gaggcaaaac agcagggccg cgccggtggc gtttttccat aggctccgcc
      121 ctcctgccag agttcacata aacagacgct tttccggtgc atctgtggga gccgtgaggc
      181 tcaaccatga atctgacagt acgggcgaaa cccgacagga cttaaagatc cccaccgttt
      241 ccggcgggtc gctccctctt gcgctctcct gttccgaccc tgccgtttac cggatacctg
      301 ttccgccttt ctcccttacg ggaagtgtgg cgctttctca tagctcacac actggtatct
      361 cggctcggtg taggtcgttc gctccaagct gggctgtaag caagaactcc ccgttcagcc
      421 cgactgctgc gccttatccg gtaactgttc acttgagtcc aacccggaaa agcacggtaa
      481 aacgccactg gcagcagcca ttggtaactg ggagttcgca gaggatttgt ttagctaaac
      541 acgcggttgc tcttgaagtg tgcgccaaag tccggctaca ctggaaggac agatttggtt
      601 gctgtgctct gcgaaagcca gttaccacgg ttaagcagtt ccccaactga cttaaccttc
      661 gatcaaacca cctccccagg tggttttttc gaaggttaag tcagttggtg aactgcttaa
      721 ccttcgatca aaccacctcc ccaggtggtt ttttcgttta cagggcaaaa gattacgcgc
      781 agaaaaaaag gatctcaaga agatcctttg atcttttcta ctgaaccgct ctagatttca
      841 gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc caggaggtca ctgcccgctt
      901 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
      961 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgacac gggcaacagc
     1021 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     1081 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     1141 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     1201 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     1261 ataccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     1321 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     1381 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     1441 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     1501 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     1561 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     1621 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     1681 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     1741 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     1801 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     1861 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     1921 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     1981 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     2041 atggtgtccg ggatctcgac gctctccctt aagaggacat ccggtcaaat aaaacgaaag
     2101 gctcagtcga aagactgggc ctttcgtttt gctgaggaga cttagggacc ctttaagact
     2161 ccttattacg cagtatgtta gcaaacgtag aaaatacata cataaaggtg gcaacatata
     2221 aaagaaacgc aaagacaccg cggaacaggt tgatcttatc gcagtcgata ctgaactcgt
     2281 aaggtttacc agcgccaaag acaaaagggc gacattcaac cgattgaggg agggaaggta
     2341 aatattgacg gaaattattc attaaaggtg aattatcacc gtcaccgact tgagccattt
     2401 gggaattaga gccagcaaaa tcaccagtag caccattacc attagcaagg ccggaaacgt
     2461 caccaatgaa accatcgata gcagcaccgt aatcagtagc gacagaatca agtttgcctt
     2521 tagcgtcaga ctgtagcgcg ttttcatcgg cattttcggt catagccccc ttattagcgt
     2581 ttgccatctt ttcataatca aaatcaccgg aaccagagcc accaccggaa ccgcctccct
     2641 cagagccgcc accctcagaa ccgccaccct cagagccacc accctcagag ccgccaccag
     2701 aaccaccacc agagccgccg ccagcattga caggaggttg aggcaggtca gacgattggc
     2761 cttgatattc acaaacgaat ggatcctcat taaagccaga atggaaagcg cagtctctga
     2821 atttaccgtt ccagtaagcg tcatacatgg cttttgatga tacaggagtg tactggtaat
     2881 aagttttaac ggggtcagtg ccttgagtaa cagtgcccgt ataaacagtt aatgccccct
     2941 gcctatttcg gaacctatta ttctgaaaca tgaaagtatt aagaggctga gactcctcaa
     3001 gagaaggatt aggattagcg gggttttgct cagtaccagg cggataagtg ccgtcgagag
     3061 ggttgatata agtatagccc ggaataggtg tatcaccgta ctcaggaggt ttagtaccgc
     3121 caccctcaga accgccaccc tcagaaccgc caccctcaga gccaccaccc tcattttcag
     3181 ggatagcaag cccaatagga acccatgtac cgtaacactg agtttcgtca ccagtacaaa
     3241 ctacaacgcc tgtagcattc cacagacagc cctcatagtt agcgtaacga tctaaagttt
     3301 tgtcgtcttt ccagacgtta gtaaatgaat tttctgtatg gggttttgct aaacaacttt
     3361 caacagtttc agcggagtga gaatagaaag gaacaactaa aggaattgcg aataataatt
     3421 ttttcatttt ttttttcctt tactgcacct gcaggtaatg ttgtcctctt gatttctgcg
     3481 ttcaggattg tcctgctctc tatcactgat agggatgaac tgttaataca atttgcgtgc
     3541 caatttttta tctttttgat ttataaagat ctgattgaag aatcaacagc aacatgccag
     3601 gatgagttag cgaattacac taacaagtgg cgaatttcat cacggagcca atgtcctcag
     3661 cgagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt aatttgatgc
     3721 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cgcaacgttc
     3781 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     3841 aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     3901 tactctcgca tggggagaac ccacactacc atcggcgcta cggcgtttca cttctgagtt
     3961 cggcatgggg tcaggtggga ccaccgcgct actgccgcca ggcaaattct gttttatcag
     4021 accgcttctg cgttctgatt taatctgtat caggctgaaa atcttctctc atccgccaaa
     4081 acagccaggg ccctactgat aaatgtgagc ggataacatt gacattgtga gcggataaca
     4141 agatactgag cacatcagca ggacgcactg acctcaacag aaacaataat aatggagtca
     4201 tgaccatgcc taggccgcgg gccgcgcgaa ttcagaagga gaatatacca tgaaccaaat
     4261 cgtgaagttc actgacgact ctggcctggc ggttcaagtt actccagacg atgttcgccg
     4321 ttatatctgt gagaacgcta ctgaaaaaga ggtgggcctc tttctgcaac tctgtcagac
     4381 tcaacgtttg aatccgtttg tgaaagacgc ttacctggtg aaatacggcg gtgctccagc
     4441 ttctatgatt acttcctatc aagtttttaa ccgtcgcgcg tgtcgtgatg ctaactatga
     4501 tggtatcaaa tctggtgtgg ttgttctgcg tgacggtgat gttgtgcata aacgtggtgc
     4561 tgcgtgctac aaaaaggcgg gtgaggagct catcggtggt tgggcggaag ttcgctttaa
     4621 ggatggccgc gagactgcgt atgctgaggt ggcgctcgac gactattcca ccggcaaatc
     4681 taattgggcg aaaatgccgg gtgttatgat cgaaaaatgc gcgaaggctg ctgcttggcg
     4741 cctcgcgttc ccggacactt ttcagggcat gtacgctgcg gaggaaatgg atcaagcgca
     4801 acagccagaa caggtgcgcg ctcaggcgga gcaaccagtg gatctccagc caatccgcga
     4861 actcttcaag ccatattgcg aacacttcgg catcactccg gctgagggta tgactgctgt
     4921 ttgtggtgcg gtgggcgctg aaggcatgca ctctatgacc gagcagcaag ctcgccgtgc
     4981 tcgcgcttgg atggaggaag aaatggctgc gccagctgtg gaagcggagt atgaggttgt
     5041 tgacgagggc gaggtgtttt aaggatccag gagaatacac atgccaattc aggtcttacc
     5101 gccacaactg gcgaaccaga ttgccgcagg tgaggtggtc gagcgacctg cgtcggtagt
     5161 caaagaacta gtgaaaaaca gcctcgatgc aggtgcgacg cgtatcgata ttgatatcga
     5221 acgcggtggg gcgaaactta tccgcattcg tgataacggc tgcggtatca aaaaagatga
     5281 gctggcgctg gcgctggctc gtcatgccac cagtaaaatc gcctctctgg acgatctcga
     5341 agccattatc agcctgggct ttcgcggtga ggcgctggcg agtatcagtt cggtttcccg
     5401 cctgacgctc acttcacgca ccgcagaaca gcaggaagcc tggcaggcct atgccgaagg
     5461 gcgcgatatg aacgtgacgg taaaaccggc ggcgcatcct gtggggacga cgctggaggt
     5521 gctggatctg ttctacaaca ccccggcgcg gcgcaaattc ctgcgcaccg agaaaaccga
     5581 atttaaccac attgatgaga tcatccgccg cattgcgctg gcgcgtttcg acgtcacgat
     5641 caacctgtcg cataacggta aaattgtgcg tcagtaccgc gcagtgccgg aaggcgggca
     5701 aaaagaacgg cgcttaggcg cgatttgcgg caccgctttt cttgaacaag cgctggcgat
     5761 tgaatggcaa cacggcgatc tcacgctacg cggctgggtg gccgatccaa atcacaccac
     5821 gcccgcactg gcagaaattc agtattgcta cgtgaacggt cgcatgatgc gcgatcgcct
     5881 gatcaatcac gcgatccgcc aggcctgcga agacaaactg ggggccgatc agcaaccggc
     5941 atttgtgttg tatctggaga tcgacccaca tcaggtggac gtcaacgtgc accccgccaa
     6001 acacgaagtg cgtttccatc agtcgcgtct ggtgcatgat tttatctatc agggcgtgct
     6061 gagcgtgcta caacagcaac tggaaacgcc gctaccgctg gacgatgaac cccaacctgc
     6121 accgcgttcc attccggaaa accgcgtggc ggcggggcgc aatcactttg cagaaccggc
     6181 agctcgtgag ccggtagctc cgcgctacac tcctgcgcca gcatcaggca gtcgtccggc
     6241 tgccccctgg ccgaatgcgc agccaggcta ccagaaacag caaggtgaag tgtatcgcca
     6301 gcttttgcaa acgcccgcgc cgatgcaaaa attaaaagcg ccggaaccgc aggaacctgc
     6361 acttgcggcg aacagtcaga gttttggtcg ggtactgact atcgtccatt ccgactgtgc
     6421 gttgctggag cgcgacggca acatttcact tttatccttg ccagtggcag aacgttggct
     6481 gcgtcaggca caattgacgc cgggtgaagc gcccgtttgc gcccagccgc tgctgattcc
     6541 gttgcggcta aaagtttctg ccgaagaaaa atcggcatta gaaaaagcgc agtctgccct
     6601 ggcggaattg ggtattgatt tccagtcaga tgcacagcat gtgaccatca gggcagtgcc
     6661 tttaccctta cgccaacaaa atttacaaat cttgattcct gaactgatag gctacctggc
     6721 gaagcagtcc gtattcgaac ctggcaatat tgcgcagtgg attgcacgaa atctgatgag
     6781 cgaacatgcg cagtggtcaa tggcacaggc cataaccctg ctggcggacg tggaacggtt
     6841 atgtccgcaa cttgtgaaaa cgccgccggg tggtctgtta caatctgttg atttacatcc
     6901 ggcgataaaa gccctgaaag atgagtgaaa aggaaaaaaa aatgaaatcc gctgaatatt
     6961 tgaacacttt tagattgaga aatctcggcc tacctgtcat gaacaatttg catgacatgt
     7021 ctaaggcgac tcgcatatct gttgaaacac ttcggttgtt aatctataca gctgattttc
     7081 gctataggat ctacactgta gaaaagaaag gcccagagaa gagaatgaga accatttacc
     7141 aaccttctcg agaacttaaa gccttacaag gatgggttct acgtaacatt ttagataaac
     7201 tgtcgtcatc tcctttttct attggatttg aaaagcacca atctattttg aataatgcta
     7261 ccccgcatat tggggcaaac tttatactga atattgattt ggaggatttt ttcccaagtt
     7321 taactgctaa caaagttttt ggagtgttcc attctcttgg ttataatcga ctaatatctt
     7381 cagttttgac aaaaatatgt tgttataaaa atctgctacc acaaggtgct ccatcatcac
     7441 ctaaattagc taatctaata tgttctaaac ttgattatcg tattcagggt tatgcaggta
     7501 gtcggggctt gatatatacg agatatgccg atgatctcac cttatctgca cagtctatga
     7561 aaaaggttgt taaagcacgt gattttttat tttctataat cccaagtgaa ggattggtta
     7621 ttaactcaaa aaaaacttgt attagtgggc ctcgtagtca gaggaaagtt acaggtttag
     7681 ttatttcaca agagaaagtt gggataggta gagaaaaata taaagaaatt agagcaaaga
     7741 tacatcatat attttgcggt aagtcttctg agatagaaca cgttagggga tggttgtcat
     7801 ttattttaag tgtggattca aaaagccata ggagattaat aacttatatt agcaaattag
     7861 aaaaaaaata tggaaagaac cctttaaata aagcgaagac ctaacattta actatgataa
     7921 gattccgtat gcgcaccctt agcgagaggt ttatcattaa ggtcaacctc tggatgttgt
     7981 ttcggcatcc tgcattgaat ctgagttact gtctgttttc ctgaattcca caacgctgga
     8041 acgcattgaa aagaatttcg taatcacaga cccgcgcctt cccgacaatc caattatttt
     8101 tgcgtccgat agcttcctgt acgaattcag gaaacccgtt ttttctgacg taagggtgcg
     8161 catacggaat ctacttaatt aacggcactc ctcagccaag tcaaaagcct ccgaccggag
     8221 gcttttgact acatgcccat ggcgtttacg ccccgccctg ccactcatcg cagtactgtt
     8281 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     8341 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccata gtgaaaacgg
     8401 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     8461 gattggctga aacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     8521 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     8581 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     8641 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaac tccggatgag
     8701 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     8761 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacatttag
     8821 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     8881 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     8941 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     9001 gccaagccaa ataggccgtc actcggtcgc tacgctccgg gcgtgaga
//
