LOCUS       sequence-492431-        7053 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  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 7053)
  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: eOYW014
FEATURES             Location/Qualifiers
     source          1..7053
                     /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]
     RBS             4228..4236
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             4243..4938
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     terminator      4943..5037
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5442..5830
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(6001..6372)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6405..6514
                     /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 gacggctagc tcagtcctag
     4141 gtacagtgct agctactggg cccaagttca cttaaaaagg agatcaacaa tgaaagcaat
     4201 tttcgtactg aaacatctta atcatgctaa ggaggttttc taatgagcga gctgatcaag
     4261 gagaacatgc acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc
     4321 acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgaagat caaggtggtc
     4381 gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagc
     4441 aaagccttca tcaaccacac ccagggcatc cccgacttct ttaagcagtc cttccctgag
     4501 ggcttcacat gggagagaat caccacatac gaagacgggg gcgtgctgac cgctacccag
     4561 gacaccagct tccagaacgg ctgcatcatc tacaacgtca agatcaacgg ggtgaacttc
     4621 ccatccaacg gccctgtgat gcagaagaaa acacgcggct gggaggccaa caccgagatg
     4681 ctgtaccccg ctgacggcgg cctgagaggc cacagccaga tggccctgaa gctcgtgggc
     4741 gggggctacc tgcactgctc cttcaagacc acatacagat ccaagaaacc cgctaagaac
     4801 ctcaagatgc ccggcttcca cttcgtggac cacagactgg aaagaatcaa ggaggccgac
     4861 aaagaaacgt acgtcgagca gcacgagatg gctgtggcca agtactgcga cctccctagc
     4921 aaactggggc acagatgaag gtgactcctg ttgatagatc cagtaatgac ctcagaactc
     4981 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatgct
     5041 tccaggatac atagattacc acaactccga gcccttccac cccaagctag cttggaggac
     5101 aaatccgccg ctaggagctt gcggccgcct taagacgcgt ggatccccgg gaatttcgac
     5161 cccacgcccc tctttaatac gacgggcaat ttgcacttca gaaaatgaag agtttgcttt
     5221 agccataaca aaagtccagt atgctttttc acagcataac tggactgatt tcagtttaca
     5281 actattctgt ctagtttaag actttattgt catagtttag atctattttg ttcagtttaa
     5341 gactttattg tccgcccaca cccgcttacg cagggcatcc atttattact caaccgtaac
     5401 cgattttgcc aggttacgcg gctggtcctc tagaattccc atgtcagccg ttaagtgttc
     5461 ctgtgtcact caaaattgct ttgagaggct ctaagggctt ctcagtgcgt tacatccctg
     5521 gcttgttgtc cacaaccgtt aaaccttaaa agctttaaaa gccttatata ttcttttttt
     5581 tcttataaaa cttaaaacct tagaggctat ttaagttgct gatttatatt aattttattg
     5641 ttcaaacatg agagcttagt acgtgaaaca tgagagctta gtacgttagc catgagagct
     5701 tagtacgtta gccatgaggg tttagttcgt taaacatgag agcttagtac gttaaacatg
     5761 agagcttagt acgtgaaaca tgagagctta gtacgtacta tcaacaggtt gaactgctga
     5821 tcttcagatc ctctacgccg gacgcatcgt ggccggatcg gggggggggg gcgctcggtc
     5881 ttgccttgct cgtcggtgat gtacttcacc agctccgcga agtcgctctt cttgatggag
     5941 cgcatgggga cgtgcttggc aatcacgcgc accccccggc cgttttagcg gctaaaaaag
     6001 tcatggctct gccctcgggc ggaccacgcc catcatgacc ttgccaagct cgtcctgctt
     6061 ctcttcgatc ttcgccagca gggcgaggat cgtggcatca ccgaaccgcg ccgtgcgcgg
     6121 gtcgtcggtg agccagagtt tcagcaggcc gcccaggcgg cccaggtcgc cattgatgcg
     6181 ggccagctcg cggacgtgct catagtccac gacgcccgtg attttgtagc cctggccgac
     6241 ggccagcagg taggccgaca ggctcatgcc ggccgccgcc gccttttcct caatcgctct
     6301 tcgttcgtct ggaaggcagt acaccttgat aggtgggctg cccttcctgg ttggcttggt
     6361 ttcatcagcc atccgcttgc cctcatctgt tacgccggcg gtagccggcc agcctcgcag
     6421 agcaggattc ccgttgagca ccgccaggtg cgaataaggg acagtgaaga aggaacaccc
     6481 gctcgcgggt gggcctactt cacctatcct gcccggctga cgccgttgga tacaccaagg
     6541 aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac
     6601 cgaaaaaatc gctataatga ccccgaagca gggttatgca gcggaaaagc gctttctcat
     6661 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6721 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6781 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6841 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6901 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6961 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7021 cagcagatta cgcgcagaaa aaaaggatct caa
//
