LOCUS       sequence-492413-        8322 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  IPTG inducible TurboRFP circuit with stronger signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8322)
  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: eOYW022
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(1494..1513)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1505..2320
                     /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             1505..2320
                     /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(1578..1597)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4034..4052
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4119..4153
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     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="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     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 ggggaaagcc acgttgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     1441 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     1501 tgttatgagc catattcaac gggaaacgtc ttgctcgagg ccgcgattaa attccaacat
     1561 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1621 aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     1681 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     1741 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     1801 tgcgatcccc gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     1861 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     1921 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     1981 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2041 gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc atggtgattt
     2101 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2161 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2221 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2281 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     2341 gtaacactgg cagagcatta cgctgacttg acgggacggc ggctttgttg aataaatcga
     2401 acttttgctg agttgaagga tcagatcacg catcttcccg acaacgcaga ccgttccgtg
     2461 gcaaagcaaa agttcaaaat caccaactgg tccacctaca acaaagctct catcaaccgt
     2521 ggctccctca ctttctggct ggatgatggg gcgattcagg cctggtatga gtcagcaaca
     2581 ccttcttcac gaggcagacc tcagcgcccc cccccccctg caggtcgact agagtcgggg
     2641 atccggtgat tgattgagca agctttatgc ttgtaaaccg ttttgtgaaa aaatttttaa
     2701 aataaaaaag gggacctcta gggtccccaa ttaattagta atataatcta ttaaaggtca
     2761 ttcaaaaggt catccaccgg atcaattccc ctgctcgcgc aggctgggtg ccaagctctc
     2821 gggtaacatc aaggcccgat ccttggagcc cttgccctcc cgcacgatga tcgtgccgtg
     2881 atcgaaatcc agatccttga cccgcagttg caaaccctca ctgatccgca tgcccgttcc
     2941 atacagaagc tgggcgaaca aacgatgctc gccttccaga aaaccgagga tgcgaaccac
     3001 ttcatccggg gtcagcacca ccggcaagcg ccgcgacggc cgaggtcttc cgatctcctg
     3061 aagccagggc agatccgtgc acagcacctt gccgtagaag aacagcaagg ccgccaatgc
     3121 ctgacgatgc gtggagaccg aaaccttgcg ctcgttcgcc agccaggaca gaaatgcctc
     3181 gacttcgctg ctgcccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg
     3241 aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc
     3301 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     3361 catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca
     3421 agcgcgttac gccgtgggtc gatgtttgtg tctaacaatt cgttcaagcc gacgccgctt
     3481 cgcggcgcgg cttaactcaa gcgttagatg cactaagcac ataattgctc acagccaaac
     3541 tatcaggtca agtctgcttt tattattttt aagcgtgcat aataagccct acacaaattg
     3601 ggagatatat catgaaaggc tggctttttc ttgttatcgc aatagttggc gaagtaatcg
     3661 caacatccgc attaaaatct agcgagggct ttactaagct gatccggtgg atgacctttt
     3721 gaatgacctt taatagatta tattactaat taattgggga ccctagaggt cccctttttt
     3781 attttaaaaa ttttttcaca aaacggttta caagcataaa gcttgctcaa tcaatcaccg
     3841 gatcccgacg ggcccggtac cgcggccgcg cgaattcgag ctcggtacca tttatcaggg
     3901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3961 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4021 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcacctcg agaacattac
     4081 tcgcatccat tctcaggctg tctcgtctcg tctcggagtt tacggctagc tcagtcctag
     4141 gtatagtgct agctactaga gtcacacagg aaagtactaa atggtgaatg tgaaaccagt
     4201 aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt
     4261 gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa 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
//
