LOCUS       sequence-346591-        4319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene expression, pCOLA origin, LacI repressor, PLlacO-1 
            promoter, mRFP, Kanamycin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4319)
  TITLE     Reisch Lab Duet Plasmids
REFERENCE   2  (bases 1 to 4319)
  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"
COMMENT     Sequence Label: pCLlR2
FEATURES             Location/Qualifiers
     source          1..4319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..46)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      42..89
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(319..1134)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(319..1134)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1042..1061
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1135..1226)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1244..1879)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(1389..1408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2077..2093
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2144..2160
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2211..3293)
                     /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     3255..3274
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3294..3371)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      complement(3435..3478)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3481..3502
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    3505..3521
                     /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)."
     protein_bind    3528..3544
                     /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             3596..3607
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3614..4291
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
ORIGIN
        1 gctttccagt cgggaaacct gctgccaccg ctgagcaata actagcataa ccccttgggg
       61 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg
      121 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
      181 aacgaccctg ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt
      241 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      301 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      361 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
      421 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
      481 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
      541 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
      601 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
      661 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca
      721 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
      781 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
      841 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
      901 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
      961 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     1021 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1081 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt
     1141 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1201 tgaatgtatt tagaaaaata aacaaatagg catgctagcg cagaaacgtc ctagaagatg
     1261 ccaggaggat acttagcaga gagacaataa ggccggagcg aagccgtttt tccataggct
     1321 ccgcccccct gacgaacatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     1381 aggactataa agataccagg cgtttccccc tgatggctcc ctcttgcgct ctcctgttcc
     1441 cgtcctgcgg cgtccgtgtt gtggtggagg ctttacccaa atcaccacgt cccgttccgt
     1501 gtagacagtt cgctccaagc tgggctgtgt gcaagaaccc cccgttcagc ccgactgctg
     1561 cgccttatcc ggtaactatc atcttgagtc caacccggaa agacacgaca aaacgccact
     1621 ggcagcagcc attggtaact gagaattagt ggatttagat atcgagagtc ttgaagtggt
     1681 ggcctaacag aggctacact gaaaggacag tatttggtat ctgcgctcca ctaaagccag
     1741 ttaccaggtt aagcagttcc ccaactgact taaccttcga tcaaaccgcc tccccaggcg
     1801 gttttttcgt ttacagagca ggagattacg acgatcgtaa aaggatctca agaagatcct
     1861 ttacggattc ccgacaccat cactctagat ttcagtgcaa tttatctctt caaatgtagc
     1921 acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc
     1981 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     2041 taatgagtga gctaacttac attaattgcg ttgcgcgtaa aacgacggcc agtgagatcc
     2101 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgcgtaaaac gacggccagt
     2161 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     2221 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2281 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     2341 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     2401 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     2461 tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg
     2521 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     2581 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     2641 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     2701 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     2761 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     2821 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     2881 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     2941 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     3001 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     3061 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     3121 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     3181 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     3241 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     3301 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     3361 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaaatggcg
     3421 tatcacgagg cagaatttca gataaaaaaa atccttagct ttcgctaagg atgatttctg
     3481 gaattcgcgg ccgcttctag agaattgtga gcggataaca attgacattg tgagcggata
     3541 acaagatact gagcacaccc tgtaggaact gggaagagcg caacacatac tagagaaaga
     3601 ggagaaatac tagatggctt cctccgaaga cgttatcaaa gagttcatgc gtttcaaagt
     3661 tcgtatggaa ggttccgtta acggtcacga gttcgaaatc gaaggtgaag gtgaaggtcg
     3721 tccgtacgaa ggtacccaga ccgctaaact gaaagttacc aaaggtggtc cgctgccgtt
     3781 cgcttgggac atcctgtccc cgcagttcca gtacggttcc aaagcttacg ttaaacaccc
     3841 ggctgacatc ccggactacc tgaaactgtc cttcccggaa ggtttcaaat gggaacgtgt
     3901 tatgaacttc gaagacggtg gtgttgttac cgttacccag gactcctccc tgcaagacgg
     3961 tgagttcatc tacaaagtta aactgcgtgg taccaacttc ccgtccgacg gtccggttat
     4021 gcagaaaaaa accatgggtt gggaagcttc caccgaacgt atgtacccgg aagacggtgc
     4081 tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac ggtggtcact acgacgctga
     4141 agttaaaacc acctacatgg ctaaaaaacc ggttcagctg ccgggtgctt acaaaaccga
     4201 catcaaactg gacatcacct cccacaacga agactacacc atcgttgaac agtacgaacg
     4261 tgctgaaggt cgtcactcca ccggtgctta ataacgctga tagtgctagt gtagatcgc
//
