LOCUS       sequence-492410-        8321 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  IPTG inducible TurboRFP circuit with medium signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8321)
  TITLE     Murray Lab Golden Gate Plasmids 2025
REFERENCE   2  (bases 1 to 8321)
  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: eOYW021
FEATURES             Location/Qualifiers
     source          1..8321
                     /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"
     RBS             4162..4173
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4189..5271
                     /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(4208..4227)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5431..5436,5437..5452,5453..5459)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5465..5484
                     /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             5493..5504
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5511..6206
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     terminator      6211..6305
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      6710..7098
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(7269..7640)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            7673..7782
                     /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 gaaagaggag aaatactaaa tggtgaatgt gaaaccagta
     4201 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     4261 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     4321 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     4381 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     4441 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     4501 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt
     4561 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     4621 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgag
     4681 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg
     4741 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat
     4801 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
     4861 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt
     4921 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt
     4981 ggtgcggata tctcggtagt gggatacgac gataccgaag atagctcatg ttatatcccg
     5041 ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg
     5101 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg
     5161 aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     5221 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagta aaggtttcag
     5281 ccaaaaaact taagaccgcc ggtcttgtcc actaccttgc agtaatgcgg tggacaggat
     5341 cggcggtttt cttttctctt ctcaagcttg cactgaaggt cctcaatcgc actggaaaca
     5401 tcaaggtcgg gagaacgatc gttggctgtg ttgacaatta atcatcggct cgtataatgt
     5461 gtggaattgt gagcgctcac aatttactag agaaagagga gaaatactaa atgagcgagc
     5521 tgatcaagga gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact
     5581 tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc atgaagatca
     5641 aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt
     5701 acggcagcaa agccttcatc aaccacaccc agggcatccc cgacttcttt aagcagtcct
     5761 tccctgaggg cttcacatgg gagagaatca ccacatacga agacgggggc gtgctgaccg
     5821 ctacccagga caccagcttc cagaacggct gcatcatcta caacgtcaag atcaacgggg
     5881 tgaacttccc atccaacggc cctgtgatgc agaagaaaac acgcggctgg gaggccaaca
     5941 ccgagatgct gtaccccgct gacggcggcc tgagaggcca cagccagatg gccctgaagc
     6001 tcgtgggcgg gggctacctg cactgctcct tcaagaccac atacagatcc aagaaacccg
     6061 ctaagaacct caagatgccc ggcttccact tcgtggacca cagactggaa agaatcaagg
     6121 aggccgacaa agaaacgtac gtcgagcagc acgagatggc tgtggccaag tactgcgacc
     6181 tccctagcaa actggggcac agatgaaggt gactcctgtt gatagatcca gtaatgacct
     6241 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
     6301 agaatgcttc caggatacat agattaccac aactccgagc ccttccaccc caagctagct
     6361 tggaggacaa atccgccgct aggagcttgc ggccgcctta agacgcgtgg atccccggga
     6421 atttcgaccc cacgcccctc tttaatacga cgggcaattt gcacttcaga aaatgaagag
     6481 tttgctttag ccataacaaa agtccagtat gctttttcac agcataactg gactgatttc
     6541 agtttacaac tattctgtct agtttaagac tttattgtca tagtttagat ctattttgtt
     6601 cagtttaaga ctttattgtc cgcccacacc cgcttacgca gggcatccat ttattactca
     6661 accgtaaccg attttgccag gttacgcggc tggtcctcta gaattcccat gtcagccgtt
     6721 aagtgttcct gtgtcactca aaattgcttt gagaggctct aagggcttct cagtgcgtta
     6781 catccctggc ttgttgtcca caaccgttaa accttaaaag ctttaaaagc cttatatatt
     6841 cttttttttc ttataaaact taaaacctta gaggctattt aagttgctga tttatattaa
     6901 ttttattgtt caaacatgag agcttagtac gtgaaacatg agagcttagt acgttagcca
     6961 tgagagctta gtacgttagc catgagggtt tagttcgtta aacatgagag cttagtacgt
     7021 taaacatgag agcttagtac gtgaaacatg agagcttagt acgtactatc aacaggttga
     7081 actgctgatc ttcagatcct ctacgccgga cgcatcgtgg ccggatcggg gggggggggc
     7141 gctcggtctt gccttgctcg tcggtgatgt acttcaccag ctccgcgaag tcgctcttct
     7201 tgatggagcg catggggacg tgcttggcaa tcacgcgcac cccccggccg ttttagcggc
     7261 taaaaaagtc atggctctgc cctcgggcgg accacgccca tcatgacctt gccaagctcg
     7321 tcctgcttct cttcgatctt cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc
     7381 gtgcgcgggt cgtcggtgag ccagagtttc agcaggccgc ccaggcggcc caggtcgcca
     7441 ttgatgcggg ccagctcgcg gacgtgctca tagtccacga cgcccgtgat tttgtagccc
     7501 tggccgacgg ccagcaggta ggccgacagg ctcatgccgg ccgccgccgc cttttcctca
     7561 atcgctcttc gttcgtctgg aaggcagtac accttgatag gtgggctgcc cttcctggtt
     7621 ggcttggttt catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag
     7681 cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag
     7741 gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata
     7801 caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat
     7861 ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaagcgc
     7921 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7981 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8041 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8101 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8161 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8221 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8281 tttgcaagca gcagattacg cgcagaaaaa aaggatctca a
//
