LOCUS       sequence-492428-        8322 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  IPTG inducible TurboRFP circuit with weak signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 4120 to 4154)
  TITLE     Murray Lab Golden Gate Plasmids 2025
REFERENCE   2  (bases 1 to 8322)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: eEML017
FEATURES             Location/Qualifiers
     source          1..8322
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     mobile_element  1210..1375
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     primer_bind     complement(1495..1514)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1506..2321
                     /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             1506..2321
                     /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     complement(1579..1598)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4035..4053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4120..4154
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             4163..4174
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4190..5272
                     /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="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTRENVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4209..4228)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5432..5437,5438..5453,5454..5460)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5466..5485
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     RBS             5494..5505
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5512..6207
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     terminator      6212..6306
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      6711..7099
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(7270..7641)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            7674..7783
                     /note="incP origin of transfer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgaattaatt cgagctcgcc caaacataac aggaagaaaa
     1201 atgccccgct gtgggcggac aaaatagttg ggaactggga ggggtggaaa tggagttttt
     1261 aaggattatt tagggaagag tgacaaaata gatgggaact gggtgtagcg tcgtaagcta
     1321 atacgaaaat taaaaatgac aaaatagttt ggaactagat ttcacttatc tggttggcct
     1381 gcaggggggg gggggaaagc cacgttgtgt ctcaaaatct ctgatgttac attgcacaag
     1441 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     1501 gtgttatgag ccatattcaa cgggaaacgt cttgctcgag gccgcgatta aattccaaca
     1561 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     1621 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     1681 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     1741 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     1801 ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     1861 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     1921 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     1981 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     2041 ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt
     2101 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     2161 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     2221 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     2281 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     2341 tgtaacactg gcagagcatt acgctgactt gacgggacgg cggctttgtt gaataaatcg
     2401 aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt
     2461 ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg
     2521 tggctccctc actttctggc tggatgatgg ggcgattcag gcctggtatg agtcagcaac
     2581 accttcttca cgaggcagac ctcagcgccc ccccccccct gcaggtcgac tagagtcggg
     2641 gatccggtga ttgattgagc aagctttatg cttgtaaacc gttttgtgaa aaaattttta
     2701 aaataaaaaa ggggacctct agggtcccca attaattagt aatataatct attaaaggtc
     2761 attcaaaagg tcatccaccg gatcaattcc cctgctcgcg caggctgggt gccaagctct
     2821 cgggtaacat caaggcccga tccttggagc ccttgccctc ccgcacgatg atcgtgccgt
     2881 gatcgaaatc cagatccttg acccgcagtt gcaaaccctc actgatccgc atgcccgttc
     2941 catacagaag ctgggcgaac aaacgatgct cgccttccag aaaaccgagg atgcgaacca
     3001 cttcatccgg ggtcagcacc accggcaagc gccgcgacgg ccgaggtctt ccgatctcct
     3061 gaagccaggg cagatccgtg cacagcacct tgccgtagaa gaacagcaag gccgccaatg
     3121 cctgacgatg cgtggagacc gaaaccttgc gctcgttcgc cagccaggac agaaatgcct
     3181 cgacttcgct gctgcccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
     3241 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
     3301 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     3361 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
     3421 aagcgcgtta cgccgtgggt cgatgtttgt gtctaacaat tcgttcaagc cgacgccgct
     3481 tcgcggcgcg gcttaactca agcgttagat gcactaagca cataattgct cacagccaaa
     3541 ctatcaggtc aagtctgctt ttattatttt taagcgtgca taataagccc tacacaaatt
     3601 gggagatata tcatgaaagg ctggcttttt cttgttatcg caatagttgg cgaagtaatc
     3661 gcaacatccg cattaaaatc tagcgagggc tttactaagc tgatccggtg gatgaccttt
     3721 tgaatgacct ttaatagatt atattactaa ttaattgggg accctagagg tccccttttt
     3781 tattttaaaa attttttcac aaaacggttt acaagcataa agcttgctca atcaatcacc
     3841 ggatcccgac gggcccggta ccgcggccgc gcgaattcga gctcggtacc atttatcagg
     3901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3961 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     4021 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcacctc gagaacatta
     4081 ctcgcatcca ttctcaggct gtctcgtctc gtctcggagt tgacggctag ctcagtccta
     4141 ggtacagtgc tagctactag agaaagagga gaaatactaa atggtgaatg tgaaaccagt
     4201 aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt
     4261 gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaac gtggaagcgg cgatggcgga
     4321 gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat
     4381 tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa
     4441 atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt
     4501 cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat
     4561 taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc
     4621 ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga
     4681 ggacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct
     4741 gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata
     4801 tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc
     4861 cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt
     4921 tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt
     4981 tggtgcggat atctcggtag tgggatacga cgataccgaa gatagctcat gttatatccc
     5041 gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt
     5101 gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt
     5161 gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     5221 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt aaaggtttca
     5281 gccaaaaaac ttaagaccgc cggtcttgtc cactaccttg cagtaatgcg gtggacagga
     5341 tcggcggttt tcttttctct tctcaagctt gcactgaagg tcctcaatcg cactggaaac
     5401 atcaaggtcg ggagaacgat cgttggctgt gttgacaatt aatcatcggc tcgtataatg
     5461 tgtggaattg tgagcgctca caatttacta gagaaagagg agaaatacta aatgagcgag
     5521 ctgatcaagg agaacatgca catgaagctg tacatggagg gcaccgtgaa caaccaccac
     5581 ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac catgaagatc
     5641 aaggtggtcg agggcggccc tctccccttc gccttcgaca tcctggctac cagcttcatg
     5701 tacggcagca aagccttcat caaccacacc cagggcatcc ccgacttctt taagcagtcc
     5761 ttccctgagg gcttcacatg ggagagaatc accacatacg aagacggggg cgtgctgacc
     5821 gctacccagg acaccagctt ccagaacggc tgcatcatct acaacgtcaa gatcaacggg
     5881 gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa cacgcggctg ggaggccaac
     5941 accgagatgc tgtaccccgc tgacggcggc ctgagaggcc acagccagat ggccctgaag
     6001 ctcgtgggcg ggggctacct gcactgctcc ttcaagacca catacagatc caagaaaccc
     6061 gctaagaacc tcaagatgcc cggcttccac ttcgtggacc acagactgga aagaatcaag
     6121 gaggccgaca aagaaacgta cgtcgagcag cacgagatgg ctgtggccaa gtactgcgac
     6181 ctccctagca aactggggca cagatgaagg tgactcctgt tgatagatcc agtaatgacc
     6241 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
     6301 gagaatgctt ccaggataca tagattacca caactccgag cccttccacc ccaagctagc
     6361 ttggaggaca aatccgccgc taggagcttg cggccgcctt aagacgcgtg gatccccggg
     6421 aatttcgacc ccacgcccct ctttaatacg acgggcaatt tgcacttcag aaaatgaaga
     6481 gtttgcttta gccataacaa aagtccagta tgctttttca cagcataact ggactgattt
     6541 cagtttacaa ctattctgtc tagtttaaga ctttattgtc atagtttaga tctattttgt
     6601 tcagtttaag actttattgt ccgcccacac ccgcttacgc agggcatcca tttattactc
     6661 aaccgtaacc gattttgcca ggttacgcgg ctggtcctct agaattccca tgtcagccgt
     6721 taagtgttcc tgtgtcactc aaaattgctt tgagaggctc taagggcttc tcagtgcgtt
     6781 acatccctgg cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag ccttatatat
     6841 tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg atttatatta
     6901 attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag tacgttagcc
     6961 atgagagctt agtacgttag ccatgagggt ttagttcgtt aaacatgaga gcttagtacg
     7021 ttaaacatga gagcttagta cgtgaaacat gagagcttag tacgtactat caacaggttg
     7081 aactgctgat cttcagatcc tctacgccgg acgcatcgtg gccggatcgg gggggggggg
     7141 cgctcggtct tgccttgctc gtcggtgatg tacttcacca gctccgcgaa gtcgctcttc
     7201 ttgatggagc gcatggggac gtgcttggca atcacgcgca ccccccggcc gttttagcgg
     7261 ctaaaaaagt catggctctg ccctcgggcg gaccacgccc atcatgacct tgccaagctc
     7321 gtcctgcttc tcttcgatct tcgccagcag ggcgaggatc gtggcatcac cgaaccgcgc
     7381 cgtgcgcggg tcgtcggtga gccagagttt cagcaggccg cccaggcggc ccaggtcgcc
     7441 attgatgcgg gccagctcgc ggacgtgctc atagtccacg acgcccgtga ttttgtagcc
     7501 ctggccgacg gccagcaggt aggccgacag gctcatgccg gccgccgccg ccttttcctc
     7561 aatcgctctt cgttcgtctg gaaggcagta caccttgata ggtgggctgc ccttcctggt
     7621 tggcttggtt tcatcagcca tccgcttgcc ctcatctgtt acgccggcgg tagccggcca
     7681 gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa
     7741 ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac gccgttggat
     7801 acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga
     7861 tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag cggaaaagcg
     7921 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7981 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     8041 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     8101 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     8161 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     8221 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     8281 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aa
//
