LOCUS       sequence-492423-        6989 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 1 to 6989)
  TITLE     Murray Lab Golden Gate Plasmids 2025
REFERENCE   2  (bases 1 to 6989)
  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: eOYW009
FEATURES             Location/Qualifiers
     source          1..6989
                     /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     4033..4051
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4118..4152
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     RBS             4161..4172
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4179..4874
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     terminator      4879..4973
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5378..5766
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(5937..6308)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6341..6450
                     /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 ccccccctgc aggtcgacta gagtcgggga
     2641 tccggtgatt gattgagcaa gctttatgct tgtaaaccgt tttgtgaaaa aatttttaaa
     2701 ataaaaaagg ggacctctag ggtccccaat taattagtaa tataatctat taaaggtcat
     2761 tcaaaaggtc atccaccgga tcaattcccc tgctcgcgca ggctgggtgc caagctctcg
     2821 ggtaacatca aggcccgatc cttggagccc ttgccctccc gcacgatgat cgtgccgtga
     2881 tcgaaatcca gatccttgac ccgcagttgc aaaccctcac tgatccgcat gcccgttcca
     2941 tacagaagct gggcgaacaa acgatgctcg ccttccagaa aaccgaggat gcgaaccact
     3001 tcatccgggg tcagcaccac cggcaagcgc cgcgacggcc gaggtcttcc gatctcctga
     3061 agccagggca gatccgtgca cagcaccttg ccgtagaaga acagcaaggc cgccaatgcc
     3121 tgacgatgcg tggagaccga aaccttgcgc tcgttcgcca gccaggacag aaatgcctcg
     3181 acttcgctgc tgcccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga
     3241 acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca
     3301 cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc
     3361 atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa
     3421 gcgcgttacg ccgtgggtcg atgtttgtgt ctaacaattc gttcaagccg acgccgcttc
     3481 gcggcgcggc ttaactcaag cgttagatgc actaagcaca taattgctca cagccaaact
     3541 atcaggtcaa gtctgctttt attattttta agcgtgcata ataagcccta cacaaattgg
     3601 gagatatatc atgaaaggct ggctttttct tgttatcgca atagttggcg aagtaatcgc
     3661 aacatccgca ttaaaatcta gcgagggctt tactaagctg atccggtgga tgaccttttg
     3721 aatgaccttt aatagattat attactaatt aattggggac cctagaggtc ccctttttta
     3781 ttttaaaaat tttttcacaa aacggtttac aagcataaag cttgctcaat caatcaccgg
     3841 atcccgacgg gcccggtacc gcggccgcgc gaattcgagc tcggtaccat ttatcagggt
     3901 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3961 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     4021 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcacctcga gaacattact
     4081 cgcatccatt ctcaggctgt ctcgtctcgt ctcggagttg acagctagct cagtcctagg
     4141 tactgtgcta gctactagag aaagaggaga aatactaaat gagcgagctg atcaaggaga
     4201 acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat
     4261 ccgagggcga aggcaagccc tacgagggca cccagaccat gaagatcaag gtggtcgagg
     4321 gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcaaag
     4381 ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc cctgagggct
     4441 tcacatggga gagaatcacc acatacgaag acgggggcgt gctgaccgct acccaggaca
     4501 ccagcttcca gaacggctgc atcatctaca acgtcaagat caacggggtg aacttcccat
     4561 ccaacggccc tgtgatgcag aagaaaacac gcggctggga ggccaacacc gagatgctgt
     4621 accccgctga cggcggcctg agaggccaca gccagatggc cctgaagctc gtgggcgggg
     4681 gctacctgca ctgctccttc aagaccacat acagatccaa gaaacccgct aagaacctca
     4741 agatgcccgg cttccacttc gtggaccaca gactggaaag aatcaaggag gccgacaaag
     4801 aaacgtacgt cgagcagcac gagatggctg tggccaagta ctgcgacctc cctagcaaac
     4861 tggggcacag atgaaggtga ctcctgttga tagatccagt aatgacctca gaactccatc
     4921 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatgcttcca
     4981 ggatacatag attaccacaa ctccgagccc ttccacccca agctagcttg gaggacaaat
     5041 ccgccgctag gagcttgcgg ccgccttaag acgcgtggat ccccgggaat ttcgacccca
     5101 cgcccctctt taatacgacg ggcaatttgc acttcagaaa atgaagagtt tgctttagcc
     5161 ataacaaaag tccagtatgc tttttcacag cataactgga ctgatttcag tttacaacta
     5221 ttctgtctag tttaagactt tattgtcata gtttagatct attttgttca gtttaagact
     5281 ttattgtccg cccacacccg cttacgcagg gcatccattt attactcaac cgtaaccgat
     5341 tttgccaggt tacgcggctg gtcctctaga attcccatgt cagccgttaa gtgttcctgt
     5401 gtcactcaaa attgctttga gaggctctaa gggcttctca gtgcgttaca tccctggctt
     5461 gttgtccaca accgttaaac cttaaaagct ttaaaagcct tatatattct tttttttctt
     5521 ataaaactta aaaccttaga ggctatttaa gttgctgatt tatattaatt ttattgttca
     5581 aacatgagag cttagtacgt gaaacatgag agcttagtac gttagccatg agagcttagt
     5641 acgttagcca tgagggttta gttcgttaaa catgagagct tagtacgtta aacatgagag
     5701 cttagtacgt gaaacatgag agcttagtac gtactatcaa caggttgaac tgctgatctt
     5761 cagatcctct acgccggacg catcgtggcc ggatcggggg gggggggcgc tcggtcttgc
     5821 cttgctcgtc ggtgatgtac ttcaccagct ccgcgaagtc gctcttcttg atggagcgca
     5881 tggggacgtg cttggcaatc acgcgcaccc cccggccgtt ttagcggcta aaaaagtcat
     5941 ggctctgccc tcgggcggac cacgcccatc atgaccttgc caagctcgtc ctgcttctct
     6001 tcgatcttcg ccagcagggc gaggatcgtg gcatcaccga accgcgccgt gcgcgggtcg
     6061 tcggtgagcc agagtttcag caggccgccc aggcggccca ggtcgccatt gatgcgggcc
     6121 agctcgcgga cgtgctcata gtccacgacg cccgtgattt tgtagccctg gccgacggcc
     6181 agcaggtagg ccgacaggct catgccggcc gccgccgcct tttcctcaat cgctcttcgt
     6241 tcgtctggaa ggcagtacac cttgataggt gggctgccct tcctggttgg cttggtttca
     6301 tcagccatcc gcttgccctc atctgttacg ccggcggtag ccggccagcc tcgcagagca
     6361 ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc
     6421 gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag
     6481 tctacacgaa ccctttggca aaatcctgta tatcgtgcga aaaaggatgg atataccgaa
     6541 aaaatcgcta taatgacccc gaagcagggt tatgcagcgg aaaagcgctt tctcatagct
     6601 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6661 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6721 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6781 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6841 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6901 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6961 agattacgcg cagaaaaaaa ggatctcaa
//
