LOCUS       sequence-347268-        4543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene expression, pACYC origin, LacI repressor, PLlacO-1 
            promoter, mRFP, Chloramphenicol Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4543)
  TITLE     Reisch Lab Duet Plasmids
REFERENCE   2  (bases 1 to 4543)
  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: pALlR1
FEATURES             Location/Qualifiers
     source          1..4543
                     /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(310..969)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     884..903
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(970..1072)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1464..1481)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1598..2143)
                     /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(1678..1697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2368..2384
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2435..3517)
                     /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     3479..3498
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3518..3595)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      complement(3659..3702)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3705..3726
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    3729..3745
                     /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             3820..3831
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3838..4515
                     /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 acgaccgggt cgaatttgct ttcgaatttc tgccattaat
      241 ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc aataactgcc
      301 ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
      361 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
      421 caccttgtcg ccttgcgtat aatatttgcc catagtgaaa acgggggcga agaagttgtc
      481 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
      541 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
      601 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
      661 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
      721 caccagctca ccgtctttca ttgccatacg gaactccgga tgagcattca tcaggcgggc
      781 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
      841 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
      901 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
      961 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     1021 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca
     1081 ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta
     1141 ttctgcgaag tgatcttccg tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc
     1201 tgccaactta ctgatttagt gtatgatggt gtttttgagg tgctccagtg gcttctgttt
     1261 ctatcagctg tccctcctgt tcagctactg acggggtggt gcgtaacggc aaaagcaccg
     1321 ccggacatca gcgctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt
     1381 cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg
     1441 tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
     1501 cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata
     1561 cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg
     1621 acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa
     1681 gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt
     1741 ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt
     1801 tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac
     1861 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca
     1921 ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg
     1981 ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg
     2041 gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc
     2101 gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa
     2161 tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt
     2221 cagccccata cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag
     2281 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     2341 agctaactta cattaattgc gttgcgcgta aaacgacggc cagtgagatc ccggtgccta
     2401 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2461 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2521 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     2581 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     2641 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     2701 atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     2761 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     2821 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     2881 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     2941 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     3001 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     3061 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     3121 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     3181 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     3241 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     3301 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     3361 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     3421 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     3481 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     3541 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     3601 tctcgacgct ctcccttatg cgactcctgc attaggaaat ggcgtatcac gaggcagaat
     3661 ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattc gcggccgctt
     3721 ctagagaatt gtgagcggat aacaattgac attgtgagtg gataacaaga tactgagcac
     3781 accctgtagg aactgggaag agcgcaacac atactagaga aagaggagaa atactagatg
     3841 gcttcctccg aagacgttat caaagagttc atgcgtttca aagttcgtat ggaaggttcc
     3901 gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta cgaaggtacc
     3961 cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg ggacatcctg
     4021 tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga catcccggac
     4081 tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa cttcgaagac
     4141 ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt catctacaaa
     4201 gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa aaaaaccatg
     4261 ggttgggaag cttccaccga acgtatgtac ccggaagacg gtgctctgaa aggtgaaatc
     4321 aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa aaccacctac
     4381 atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa actggacatc
     4441 acctcccaca acgaagacta caccatcgtt gaacagtacg aacgtgctga aggtcgtcac
     4501 tccaccggtg cttaataacg ctgatagtgc tagtgtagat cgc
//
