LOCUS       sequence-485902-        6988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TurboRFP constitutive Golden Gate construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 4118 to 4152)
  AUTHORS   Larsson EM, Wang OY, Murray RM
  TITLE     A DNA Part Library for Reliable Engineering of the Emerging Model 
            Nematode Symbiotic Bacterium Xenorhabdus griffiniae HGB2511.
  JOURNAL   ACS Synth Biol. 2025 Oct 17;14(10):4122-4126. doi: 
            10.1021/acssynbio.5c00414. Epub 2025 Sep 25.
  PUBMED    40998411
REFERENCE   2  (bases 1 to 6988)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.5c00414"; journalName: "ACS Synth Biol"; date: 
            "2025-10-17- 17"; volume: "14"; issue: "10"; pages: "4122-4126"
COMMENT     Sequence Label: eOYW013
FEATURES             Location/Qualifiers
     source          1..6988
                     /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_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     CDS             4178..4873
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     terminator      4878..4972
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5377..5765
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(5936..6307)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6340..6449
                     /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 acggctagct cagtcctagg
     4141 tacagtgcta gctactagag tcacacagga ctactaaatg agcgagctga tcaaggagaa
     4201 catgcacatg aagctgtaca tggagggcac cgtgaacaac caccacttca agtgcacatc
     4261 cgagggcgaa ggcaagccct acgagggcac ccagaccatg aagatcaagg tggtcgaggg
     4321 cggccctctc cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcaaagc
     4381 cttcatcaac cacacccagg gcatccccga cttctttaag cagtccttcc ctgagggctt
     4441 cacatgggag agaatcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac
     4501 cagcttccag aacggctgca tcatctacaa cgtcaagatc aacggggtga acttcccatc
     4561 caacggccct gtgatgcaga agaaaacacg cggctgggag gccaacaccg agatgctgta
     4621 ccccgctgac ggcggcctga gaggccacag ccagatggcc ctgaagctcg tgggcggggg
     4681 ctacctgcac tgctccttca agaccacata cagatccaag aaacccgcta agaacctcaa
     4741 gatgcccggc ttccacttcg tggaccacag actggaaaga atcaaggagg ccgacaaaga
     4801 aacgtacgtc gagcagcacg agatggctgt ggccaagtac tgcgacctcc ctagcaaact
     4861 ggggcacaga tgaaggtgac tcctgttgat agatccagta atgacctcag aactccatct
     4921 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga atgcttccag
     4981 gatacataga ttaccacaac tccgagccct tccaccccaa gctagcttgg aggacaaatc
     5041 cgccgctagg agcttgcggc cgccttaaga cgcgtggatc cccgggaatt tcgaccccac
     5101 gcccctcttt aatacgacgg gcaatttgca cttcagaaaa tgaagagttt gctttagcca
     5161 taacaaaagt ccagtatgct ttttcacagc ataactggac tgatttcagt ttacaactat
     5221 tctgtctagt ttaagacttt attgtcatag tttagatcta ttttgttcag tttaagactt
     5281 tattgtccgc ccacacccgc ttacgcaggg catccattta ttactcaacc gtaaccgatt
     5341 ttgccaggtt acgcggctgg tcctctagaa ttcccatgtc agccgttaag tgttcctgtg
     5401 tcactcaaaa ttgctttgag aggctctaag ggcttctcag tgcgttacat ccctggcttg
     5461 ttgtccacaa ccgttaaacc ttaaaagctt taaaagcctt atatattctt ttttttctta
     5521 taaaacttaa aaccttagag gctatttaag ttgctgattt atattaattt tattgttcaa
     5581 acatgagagc ttagtacgtg aaacatgaga gcttagtacg ttagccatga gagcttagta
     5641 cgttagccat gagggtttag ttcgttaaac atgagagctt agtacgttaa acatgagagc
     5701 ttagtacgtg aaacatgaga gcttagtacg tactatcaac aggttgaact gctgatcttc
     5761 agatcctcta cgccggacgc atcgtggccg gatcgggggg ggggggcgct cggtcttgcc
     5821 ttgctcgtcg gtgatgtact tcaccagctc cgcgaagtcg ctcttcttga tggagcgcat
     5881 ggggacgtgc ttggcaatca cgcgcacccc ccggccgttt tagcggctaa aaaagtcatg
     5941 gctctgccct cgggcggacc acgcccatca tgaccttgcc aagctcgtcc tgcttctctt
     6001 cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg cgcgggtcgt
     6061 cggtgagcca gagtttcagc aggccgccca ggcggcccag gtcgccattg atgcgggcca
     6121 gctcgcggac gtgctcatag tccacgacgc ccgtgatttt gtagccctgg ccgacggcca
     6181 gcaggtaggc cgacaggctc atgccggccg ccgccgcctt ttcctcaatc gctcttcgtt
     6241 cgtctggaag gcagtacacc ttgataggtg ggctgccctt cctggttggc ttggtttcat
     6301 cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct cgcagagcag
     6361 gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg
     6421 cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt
     6481 ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa
     6541 aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaagcgcttt ctcatagctc
     6601 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6661 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6721 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6781 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6841 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6901 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6961 gattacgcgc agaaaaaaag gatctcaa
//
