LOCUS       sequence-342863-        5683 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Inducible gene expression, pET origin, LacI repressor, PLlacO-1 
            promoter, mRFP, Carbenicillin Resistance Marker. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5683)
  TITLE     Reisch Lab Duet Plasmids
REFERENCE   2  (bases 1 to 5683)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pELlR4
FEATURES             Location/Qualifiers
     source          1..5683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1..48
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      85..540
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(172..191)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     382..403
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(655..1446,1447..1515))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRKQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1278..1297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1516..1608)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1689..2277
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2178..2197
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2431..2448
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2463..2602
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2528..2547)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2688..2710
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2704..2895)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3467..3483
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3534..4616)
                     /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     4578..4597
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4617..4694)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      complement(4758..4801)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4804..4825
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    4828..4844
                     /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    4851..4867
                     /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             4919..4930
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4937..5614
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(join(5670..5683,1..5))
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
       61 actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
      121 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
      181 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
      241 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
      301 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
      361 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
      421 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
      481 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
      541 ctggcggcac gatggcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      601 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      661 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
      721 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
      781 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
      841 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
      901 taattgtttc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
      961 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1021 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1081 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1141 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1201 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1261 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1321 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1381 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1441 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1501 atgttgaata ctcatactct tcctttttca atcatgattg aagcatttat cagggttatt
     1561 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggtcatgacc
     1621 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1681 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1741 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1801 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     1861 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1921 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1981 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2041 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2101 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2161 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2221 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2281 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2341 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2401 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2461 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     2521 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     2581 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     2641 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     2701 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     2761 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     2821 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     2881 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     2941 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     3001 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     3061 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     3121 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     3181 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     3241 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     3301 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat
     3361 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     3421 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgcgtaa aacgacggcc
     3481 agtgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     3541 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3601 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     3661 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     3721 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     3781 ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac
     3841 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     3901 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     3961 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     4021 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     4081 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     4141 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     4201 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     4261 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     4321 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     4381 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     4441 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     4501 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     4561 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     4621 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     4681 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaaatg
     4741 gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt
     4801 ctggaattcg cggccgcttc tagagaattg tgagcggata acaattgaca ttgtgagcgg
     4861 ataacaagat actgagcaca ccctgtagga actgggaaga gcgcaacaca tactagagaa
     4921 agaggagaaa tactagatgg cttcctccga agacgttatc aaagagttca tgcgtttcaa
     4981 agttcgtatg gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg aaggtgaagg
     5041 tcgtccgtac gaaggtaccc agaccgctaa actgaaagtt accaaaggtg gtccgctgcc
     5101 gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt tccaaagctt acgttaaaca
     5161 cccggctgac atcccggact acctgaaact gtccttcccg gaaggtttca aatgggaacg
     5221 tgttatgaac ttcgaagacg gtggtgttgt taccgttacc caggactcct ccctgcaaga
     5281 cggtgagttc atctacaaag ttaaactgcg tggtaccaac ttcccgtccg acggtccggt
     5341 tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc cggaagacgg
     5401 tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc actacgacgc
     5461 tgaagttaaa accacctaca tggctaaaaa accggttcag ctgccgggtg cttacaaaac
     5521 cgacatcaaa ctggacatca cctcccacaa cgaagactac accatcgttg aacagtacga
     5581 acgtgctgaa ggtcgtcact ccaccggtgc ttaataacgc tgatagtgct agtgtagatc
     5641 gcgctttcca gtcgggaaac ctgctgccac cgctgagcaa taa
//
