LOCUS       sequence-492443-        6996 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 4120 to 4154)
  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 6996)
  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: eOYW018
FEATURES             Location/Qualifiers
     source          1..6996
                     /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(1495..1514)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1506..2321
                     /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             1506..2321
                     /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(1579..1598)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4035..4053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4120..4154
                     /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             4168..4179
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4187..4882
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     terminator      4887..4981
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5386..5774
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(5944..6315)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6348..6457
                     /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 gggggaaagc cacgttgtgt ctcaaaatct ctgatgttac attgcacaag
     1441 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     1501 gtgttatgag ccatattcaa cgggaaacgt cttgctcgag gccgcgatta aattccaaca
     1561 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     1621 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     1681 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     1741 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     1801 ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     1861 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     1921 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     1981 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     2041 ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt
     2101 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     2161 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     2221 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     2281 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     2341 tgtaacactg gcagagcatt acgctgactt gacgggacgg cggctttgtt gaataaatcg
     2401 aacttttgct gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt
     2461 ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg
     2521 tggctccctc actttctggc tggatgatgg ggcgattcag gcctggtatg agtcagcaac
     2581 accttcttca cgaggcagac ctcagcgccc ccccccccct gcaggtcgac tagagtcggg
     2641 gatccggtga ttgattgagc aagctttatg cttgtaaacc gttttgtgaa aaaattttta
     2701 aaataaaaaa ggggacctct agggtcccca attaattagt aatataatct attaaaggtc
     2761 attcaaaagg tcatccaccg gatcaattcc cctgctcgcg caggctgggt gccaagctct
     2821 cgggtaacat caaggcccga tccttggagc ccttgccctc ccgcacgatg atcgtgccgt
     2881 gatcgaaatc cagatccttg acccgcagtt gcaaaccctc actgatccgc atgcccgttc
     2941 catacagaag ctgggcgaac aaacgatgct cgccttccag aaaaccgagg atgcgaacca
     3001 cttcatccgg ggtcagcacc accggcaagc gccgcgacgg ccgaggtctt ccgatctcct
     3061 gaagccaggg cagatccgtg cacagcacct tgccgtagaa gaacagcaag gccgccaatg
     3121 cctgacgatg cgtggagacc gaaaccttgc gctcgttcgc cagccaggac agaaatgcct
     3181 cgacttcgct gctgcccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
     3241 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
     3301 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     3361 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
     3421 aagcgcgtta cgccgtgggt cgatgtttgt gtctaacaat tcgttcaagc cgacgccgct
     3481 tcgcggcgcg gcttaactca agcgttagat gcactaagca cataattgct cacagccaaa
     3541 ctatcaggtc aagtctgctt ttattatttt taagcgtgca taataagccc tacacaaatt
     3601 gggagatata tcatgaaagg ctggcttttt cttgttatcg caatagttgg cgaagtaatc
     3661 gcaacatccg cattaaaatc tagcgagggc tttactaagc tgatccggtg gatgaccttt
     3721 tgaatgacct ttaatagatt atattactaa ttaattgggg accctagagg tccccttttt
     3781 tattttaaaa attttttcac aaaacggttt acaagcataa agcttgctca atcaatcacc
     3841 ggatcccgac gggcccggta ccgcggccgc gcgaattcga gctcggtacc atttatcagg
     3901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3961 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     4021 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcacctc gagaacatta
     4081 ctcgcatcca ttctcaggct gtctcgtctc gtctcggagt tgacggctag ctcagtccta
     4141 ggtacagtgc tagctactga attcattaaa gaggagaaag gtaccaatga gcgagctgat
     4201 caaggagaac atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa
     4261 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga agatcaaggt
     4321 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg
     4381 cagcaaagcc ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc
     4441 tgagggcttc acatgggaga gaatcaccac atacgaagac gggggcgtgc tgaccgctac
     4501 ccaggacacc agcttccaga acggctgcat catctacaac gtcaagatca acggggtgaa
     4561 cttcccatcc aacggccctg tgatgcagaa gaaaacacgc ggctgggagg ccaacaccga
     4621 gatgctgtac cccgctgacg gcggcctgag aggccacagc cagatggccc tgaagctcgt
     4681 gggcgggggc tacctgcact gctccttcaa gaccacatac agatccaaga aacccgctaa
     4741 gaacctcaag atgcccggct tccacttcgt ggaccacaga ctggaaagaa tcaaggaggc
     4801 cgacaaagaa acgtacgtcg agcagcacga gatggctgtg gccaagtact gcgacctccc
     4861 tagcaaactg gggcacagat gaaggtgact cctgttgata gatccagtaa tgacctcaga
     4921 actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa
     4981 tgcttccagg atacatagat taccacaact ccgagccctt ccaccccaag ctagcttgga
     5041 ggacaaatcc gccgctagga gcttgcggcc gccttaagac gcgtggatcc ccgggaattt
     5101 cgaccccacg cccctcttta atacgacggg caatttgcac ttcagaaaat gaagagtttg
     5161 ctttagccat aacaaaagtc cagtatgctt tttcacagca taactggact gatttcagtt
     5221 tacaactatt ctgtctagtt taagacttta ttgtcatagt ttagatctat tttgttcagt
     5281 ttaagacttt attgtccgcc cacacccgct tacgcagggc atccatttat tactcaaccg
     5341 taaccgattt tgccaggtta cgcggctggt cctctagaat tcccatgtca gccgttaagt
     5401 gttcctgtgt cactcaaaat tgctttgaga ggctctaagg gcttctcagt gcgttacatc
     5461 cctggcttgt tgtccacaac cgttaaacct taaaagcttt aaaagcctta tatattcttt
     5521 tttttcttat aaaacttaaa accttagagg ctatttaagt tgctgattta tattaatttt
     5581 attgttcaaa catgagagct tagtacgtga aacatgagag cttagtacgt tagccatgag
     5641 agcttagtac gttagccatg agggtttagt tcgttaaaca tgagagctta gtacgttaaa
     5701 catgagagct tagtacgtga aacatgagag cttagtacgt actatcaaca ggttgaactg
     5761 ctgatcttca gatcctctac gccggacgca tcgtggccgg atcggggggg ggggcgctcg
     5821 gtcttgcctt gctcgtcggt gatgtacttc accagctccg cgaagtcgct cttcttgatg
     5881 gagcgcatgg ggacgtgctt ggcaatcacg cgcacccccc ggccgtttta gcggctaaaa
     5941 aagtcatggc tctgccctcg ggcggaccac gcccatcatg accttgccaa gctcgtcctg
     6001 cttctcttcg atcttcgcca gcagggcgag gatcgtggca tcaccgaacc gcgccgtgcg
     6061 cgggtcgtcg gtgagccaga gtttcagcag gccgcccagg cggcccaggt cgccattgat
     6121 gcgggccagc tcgcggacgt gctcatagtc cacgacgccc gtgattttgt agccctggcc
     6181 gacggccagc aggtaggccg acaggctcat gccggccgcc gccgcctttt cctcaatcgc
     6241 tcttcgttcg tctggaaggc agtacacctt gataggtggg ctgcccttcc tggttggctt
     6301 ggtttcatca gccatccgct tgccctcatc tgttacgccg gcggtagccg gccagcctcg
     6361 cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca
     6421 cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca
     6481 aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata
     6541 taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa agcgctttct
     6601 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6661 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6721 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6781 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6841 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6901 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6961 aagcagcaga ttacgcgcag aaaaaaagga tctcaa
//
