LOCUS       sequence-492374-        7051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFPmut3 constitutive Golden Gate construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 4154 to 4188)
  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 7051)
  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: eEML006
FEATURES             Location/Qualifiers
     source          1..7051
                     /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)"
     CDS             1566..2357
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     4069..4087
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4154..4188
                     /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             4197..4208
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4218..4931
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(4248..4276)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4871..4891
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      4942..5036
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5441..5829
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(5999..6370)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6403..6512
                     /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 tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa
     1561 acatcatgag ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg
     1621 tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg
     1681 atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg
     1741 atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag
     1801 agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt
     1861 ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg
     1921 caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa
     1981 gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg
     2041 aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact
     2101 gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa
     2161 ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc
     2221 agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct
     2281 tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca
     2341 aggtagtcgg caaataatca gaattggtta attggttgta acactggcag agcattacgc
     2401 tgacttgacg ggacggcggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca
     2461 gatcacgcat cttcccgaca acgcagaccg ttccgtggca aagcaaaagt tcaaaatcac
     2521 caactggtcc acctacaaca aagctctcat caaccgtggc tccctcactt tctggctgga
     2581 tgatggggcg attcaggcct ggtatgagtc agcaacacct tcttcacgag gcagacctca
     2641 gcgccccccc ccctgcaggt cgactagagt cggggatccg gtgattgatt gagcaagctt
     2701 tatgcttgta aaccgttttg tgaaaaaatt tttaaaataa aaaaggggac ctctagggtc
     2761 cccaattaat tagtaatata atctattaaa ggtcattcaa aaggtcatcc accggatcaa
     2821 ttcccctgct cgcgcaggct gggtgccaag ctctcgggta acatcaaggc ccgatccttg
     2881 gagcccttgc cctcccgcac gatgatcgtg ccgtgatcga aatccagatc cttgacccgc
     2941 agttgcaaac cctcactgat ccgcatgccc gttccataca gaagctgggc gaacaaacga
     3001 tgctcgcctt ccagaaaacc gaggatgcga accacttcat ccggggtcag caccaccggc
     3061 aagcgccgcg acggccgagg tcttccgatc tcctgaagcc agggcagatc cgtgcacagc
     3121 accttgccgt agaagaacag caaggccgcc aatgcctgac gatgcgtgga gaccgaaacc
     3181 ttgcgctcgt tcgccagcca ggacagaaat gcctcgactt cgctgctgcc caaggttgcc
     3241 gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg
     3301 gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc
     3361 gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
     3421 ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
     3481 ttgtgtctaa caattcgttc aagccgacgc cgcttcgcgg cgcggcttaa ctcaagcgtt
     3541 agatgcacta agcacataat tgctcacagc caaactatca ggtcaagtct gcttttatta
     3601 tttttaagcg tgcataataa gccctacaca aattgggaga tatatcatga aaggctggct
     3661 ttttcttgtt atcgcaatag ttggcgaagt aatcgcaaca tccgcattaa aatctagcga
     3721 gggctttact aagctgatcc ggtggatgac cttttgaatg acctttaata gattatatta
     3781 ctaattaatt ggggacccta gaggtcccct tttttatttt aaaaattttt tcacaaaacg
     3841 gtttacaagc ataaagcttg ctcaatcaat caccggatcc cgacgggccc ggtaccgcgg
     3901 ccgcgcgaat tcgagctcgg taccatttat cagggttatt gtctcatgag cggatacata
     3961 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4021 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4081 acgaggccct ttcgtcttca cctcgagaac attactcgca tccattctca ggctgtctcg
     4141 tctcgtctcg gagttgacgg ctagctcagt cctaggtaca gtgctagcta ctagagaaag
     4201 aggagaaata ctaaatgatg cgtaaaggag aagaactttt cactggagtt gtcccaattc
     4261 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag
     4321 gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg
     4381 ttccatggcc aacacttgtc actactttcg gttatggtgt tcaatgcttt gcgagatacc
     4441 cagatcatat gaaacagcat gactttttca agagtgccat gcccgaaggt tatgtacagg
     4501 aaagaactat atttttcaaa gatgacggga actacaagac acgtgctgaa gtcaagtttg
     4561 aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa
     4621 acattcttgg acacaaattg gaatacaact ataactcaca caatgtatac atcatggcag
     4681 acaaacaaaa gaatggaatc aaagttaact tcaaaattag acacaacatt gaagatggaa
     4741 gcgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
     4801 taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga
     4861 gagatcacat ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg
     4921 aactatacaa ataataaagg tgactcctgt tgatagatcc agtaatgacc tcagaactcc
     4981 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatgctt
     5041 ccaggataca tagattacca caactccgag cccttccacc ccaagctagc ttggaggaca
     5101 aatccgccgc taggagcttg cggccgcctt aagacgcgtg gatccccggg aatttcgacc
     5161 ccacgcccct ctttaatacg acgggcaatt tgcacttcag aaaatgaaga gtttgcttta
     5221 gccataacaa aagtccagta tgctttttca cagcataact ggactgattt cagtttacaa
     5281 ctattctgtc tagtttaaga ctttattgtc atagtttaga tctattttgt tcagtttaag
     5341 actttattgt ccgcccacac ccgcttacgc agggcatcca tttattactc aaccgtaacc
     5401 gattttgcca ggttacgcgg ctggtcctct agaattccca tgtcagccgt taagtgttcc
     5461 tgtgtcactc aaaattgctt tgagaggctc taagggcttc tcagtgcgtt acatccctgg
     5521 cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag ccttatatat tctttttttt
     5581 cttataaaac ttaaaacctt agaggctatt taagttgctg atttatatta attttattgt
     5641 tcaaacatga gagcttagta cgtgaaacat gagagcttag tacgttagcc atgagagctt
     5701 agtacgttag ccatgagggt ttagttcgtt aaacatgaga gcttagtacg ttaaacatga
     5761 gagcttagta cgtgaaacat gagagcttag tacgtactat caacaggttg aactgctgat
     5821 cttcagatcc tctacgccgg acgcatcgtg gccggatcgg gggggggggc gctcggtctt
     5881 gccttgctcg tcggtgatgt acttcaccag ctccgcgaag tcgctcttct tgatggagcg
     5941 catggggacg tgcttggcaa tcacgcgcac cccccggccg ttttagcggc taaaaaagtc
     6001 atggctctgc cctcgggcgg accacgccca tcatgacctt gccaagctcg tcctgcttct
     6061 cttcgatctt cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc gtgcgcgggt
     6121 cgtcggtgag ccagagtttc agcaggccgc ccaggcggcc caggtcgcca ttgatgcggg
     6181 ccagctcgcg gacgtgctca tagtccacga cgcccgtgat tttgtagccc tggccgacgg
     6241 ccagcaggta ggccgacagg ctcatgccgg ccgccgccgc cttttcctca atcgctcttc
     6301 gttcgtctgg aaggcagtac accttgatag gtgggctgcc cttcctggtt ggcttggttt
     6361 catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag
     6421 caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc
     6481 tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa
     6541 agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg
     6601 aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaagcgc tttctcatag
     6661 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6721 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6781 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6841 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6901 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6961 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7021 gcagattacg cgcagaaaaa aaggatctca a
//
