LOCUS       sequence-492435-        7078 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  Strongest TurboRFP constitutive Golden Gate construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 4119 to 4153)
  TITLE     Murray Lab Golden Gate Plasmids 2025
REFERENCE   2  (bases 1 to 7078)
  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: eOYW020
FEATURES             Location/Qualifiers
     source          1..7078
                     /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_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     misc_RNA        4160..4209
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             4239..4261
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4268..4963
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     terminator      4968..5062
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5467..5855
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(6026..6397)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6430..6539
                     /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 aagttgtgtc 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 gacggctagc tcagtcctag
     4141 gtacagtgct agctactagc tgtcaccgga tgtgctttcc ggtctgatga gtccgtgagg
     4201 acgaaacagc ctctacaaat aattttgttt aaaataattt tgtttaactt taagaaggag
     4261 atatacaatg agcgagctga tcaaggagaa catgcacatg aagctgtaca tggagggcac
     4321 cgtgaacaac caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac
     4381 ccagaccatg aagatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct
     4441 ggctaccagc ttcatgtacg gcagcaaagc cttcatcaac cacacccagg gcatccccga
     4501 cttctttaag cagtccttcc ctgagggctt cacatgggag agaatcacca catacgaaga
     4561 cgggggcgtg ctgaccgcta cccaggacac cagcttccag aacggctgca tcatctacaa
     4621 cgtcaagatc aacggggtga acttcccatc caacggccct gtgatgcaga agaaaacacg
     4681 cggctgggag gccaacaccg agatgctgta ccccgctgac ggcggcctga gaggccacag
     4741 ccagatggcc ctgaagctcg tgggcggggg ctacctgcac tgctccttca agaccacata
     4801 cagatccaag aaacccgcta agaacctcaa gatgcccggc ttccacttcg tggaccacag
     4861 actggaaaga atcaaggagg ccgacaaaga aacgtacgtc gagcagcacg agatggctgt
     4921 ggccaagtac tgcgacctcc ctagcaaact ggggcacaga tgaaggtgac tcctgttgat
     4981 agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg
     5041 ggcgtttttt attggtgaga atgcttccag gatacataga ttaccacaac tccgagccct
     5101 tccaccccaa gctagcttgg aggacaaatc cgccgctagg agcttgcggc cgccttaaga
     5161 cgcgtggatc cccgggaatt tcgaccccac gcccctcttt aatacgacgg gcaatttgca
     5221 cttcagaaaa tgaagagttt gctttagcca taacaaaagt ccagtatgct ttttcacagc
     5281 ataactggac tgatttcagt ttacaactat tctgtctagt ttaagacttt attgtcatag
     5341 tttagatcta ttttgttcag tttaagactt tattgtccgc ccacacccgc ttacgcaggg
     5401 catccattta ttactcaacc gtaaccgatt ttgccaggtt acgcggctgg tcctctagaa
     5461 ttcccatgtc agccgttaag tgttcctgtg tcactcaaaa ttgctttgag aggctctaag
     5521 ggcttctcag tgcgttacat ccctggcttg ttgtccacaa ccgttaaacc ttaaaagctt
     5581 taaaagcctt atatattctt ttttttctta taaaacttaa aaccttagag gctatttaag
     5641 ttgctgattt atattaattt tattgttcaa acatgagagc ttagtacgtg aaacatgaga
     5701 gcttagtacg ttagccatga gagcttagta cgttagccat gagggtttag ttcgttaaac
     5761 atgagagctt agtacgttaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg
     5821 tactatcaac aggttgaact gctgatcttc agatcctcta cgccggacgc atcgtggccg
     5881 gatcgggggg ggggggcgct cggtcttgcc ttgctcgtcg gtgatgtact tcaccagctc
     5941 cgcgaagtcg ctcttcttga tggagcgcat ggggacgtgc ttggcaatca cgcgcacccc
     6001 ccggccgttt tagcggctaa aaaagtcatg gctctgccct cgggcggacc acgcccatca
     6061 tgaccttgcc aagctcgtcc tgcttctctt cgatcttcgc cagcagggcg aggatcgtgg
     6121 catcaccgaa ccgcgccgtg cgcgggtcgt cggtgagcca gagtttcagc aggccgccca
     6181 ggcggcccag gtcgccattg atgcgggcca gctcgcggac gtgctcatag tccacgacgc
     6241 ccgtgatttt gtagccctgg ccgacggcca gcaggtaggc cgacaggctc atgccggccg
     6301 ccgccgcctt ttcctcaatc gctcttcgtt cgtctggaag gcagtacacc ttgataggtg
     6361 ggctgccctt cctggttggc ttggtttcat cagccatccg cttgccctca tctgttacgc
     6421 cggcggtagc cggccagcct cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat
     6481 aagggacagt gaagaaggaa cacccgctcg cgggtgggcc tacttcacct atcctgcccg
     6541 gctgacgccg ttggatacac caaggaaagt ctacacgaac cctttggcaa aatcctgtat
     6601 atcgtgcgaa aaaggatgga tataccgaaa aaatcgctat aatgaccccg aagcagggtt
     6661 atgcagcgga aaagcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6721 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6781 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6841 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6901 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6961 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7021 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaa
//
