LOCUS       sequence-474312-        6952 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 4054 to 4088)
  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 6952)
  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: eEML009
FEATURES             Location/Qualifiers
     source          1..6952
                     /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)"
     promoter        join(1505..1510,1511..1527,1528..1533)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1722..2255
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAK"
     primer_bind     3969..3987
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4054..4088
                     /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             4097..4108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4118..4831
                     /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(4148..4176)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4771..4791
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      4842..4936
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5341..5729
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(5900..6271)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6304..6413
                     /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 tgttttgaca taagcctgtt cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg
     1561 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     1621 ggcttgttat gactgttttt ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     1681 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagcaacg
     1741 atgttacgca gcagggcagt cgccctaaaa caaagttagg tggctcaagt atgggcatca
     1801 ttcgcacatg taggctcggc cctgaccaag tcaaatccat gcgggctgct cttgatcttt
     1861 tcggtcgtga gttcggagac gtagccacct actcccaaca tcagccggac tccgattacc
     1921 tcgggaactt gctccgtagt aagacattca tcgcgcttgc tgccttcgac caagaagcgg
     1981 ttgttggcgc tctcgcggct tacgttctgc ccaagtttga gcagccgcgt agtgagatct
     2041 atatctatga tctcgcagtc tccggcgagc accggaggca gggcattgcc accgcgctca
     2101 tcaatctcct caagcatgag gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag
     2161 attacggtga cgatcccgca gtggctctct atacaaagtt gggcatacgg gaagaagtga
     2221 tgcactttga tatcgaccca agtaccgcca aataatcaga attggttaat tggttgtaac
     2281 actggcagag cattacgctg acttgacggg acggcggctt tgttgaataa atcgaacttt
     2341 tgctgagttg aaggatcaga tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa
     2401 gcaaaagttc aaaatcacca actggtccac ctacaacaaa gctctcatca accgtggctc
     2461 cctcactttc tggctggatg atggggcgat tcaggcctgg tatgagtcag caacaccttc
     2521 ttcacgaggc agacctcagc gccccccccc ccctgcaggt cgactagagt cggggatccg
     2581 gtgattgatt gagcaagctt tatgcttgta aaccgttttg tgaaaaaatt tttaaaataa
     2641 aaaaggggac ctctagggtc cccaattaat tagtaatata atctattaaa ggtcattcaa
     2701 aaggtcatcc accggatcaa ttcccctgct cgcgcaggct gggtgccaag ctctcgggta
     2761 acatcaaggc ccgatccttg gagcccttgc cctcccgcac gatgatcgtg ccgtgatcga
     2821 aatccagatc cttgacccgc agttgcaaac cctcactgat ccgcatgccc gttccataca
     2881 gaagctgggc gaacaaacga tgctcgcctt ccagaaaacc gaggatgcga accacttcat
     2941 ccggggtcag caccaccggc aagcgccgcg acggccgagg tcttccgatc tcctgaagcc
     3001 agggcagatc cgtgcacagc accttgccgt agaagaacag caaggccgcc aatgcctgac
     3061 gatgcgtgga gaccgaaacc ttgcgctcgt tcgccagcca ggacagaaat gcctcgactt
     3121 cgctgctgcc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     3181 agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca
     3241 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     3301 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
     3361 gttacgccgt gggtcgatgt ttgtgtctaa caattcgttc aagccgacgc cgcttcgcgg
     3421 cgcggcttaa ctcaagcgtt agatgcacta agcacataat tgctcacagc caaactatca
     3481 ggtcaagtct gcttttatta tttttaagcg tgcataataa gccctacaca aattgggaga
     3541 tatatcatga aaggctggct ttttcttgtt atcgcaatag ttggcgaagt aatcgcaaca
     3601 tccgcattaa aatctagcga gggctttact aagctgatcc ggtggatgac cttttgaatg
     3661 acctttaata gattatatta ctaattaatt ggggacccta gaggtcccct tttttatttt
     3721 aaaaattttt tcacaaaacg gtttacaagc ataaagcttg ctcaatcaat caccggatcc
     3781 cgacgggccc ggtaccgcgg ccgcgcgaat tcgagctcgg taccatttat cagggttatt
     3841 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3901 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     3961 cctataaaaa taggcgtatc acgaggccct ttcgtcttca cctcgagaac attactcgca
     4021 tccattctca ggctgtctcg tctcgtctcg gagttgacgg ctagctcagt cctaggtaca
     4081 gtgctagcta ctagagaaag aggagaaata ctaaatgatg cgtaaaggag aagaactttt
     4141 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc
     4201 tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg
     4261 cactactgga aaactacctg ttccatggcc aacacttgtc actactttcg gttatggtgt
     4321 tcaatgcttt gcgagatacc cagatcatat gaaacagcat gactttttca agagtgccat
     4381 gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac
     4441 acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat
     4501 tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact ataactcaca
     4561 caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagttaact tcaaaattag
     4621 acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat
     4681 tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc
     4741 gaaagatccc aacgaaaaga gagatcacat ggtccttctt gagtttgtaa cagctgctgg
     4801 gattacacat ggcatggatg aactatacaa ataataaagg tgactcctgt tgatagatcc
     4861 agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt
     4921 ttttattggt gagaatgctt ccaggataca tagattacca caactccgag cccttccacc
     4981 ccaagctagc ttggaggaca aatccgccgc taggagcttg cggccgcctt aagacgcgtg
     5041 gatccccggg aatttcgacc ccacgcccct ctttaatacg acgggcaatt tgcacttcag
     5101 aaaatgaaga gtttgcttta gccataacaa aagtccagta tgctttttca cagcataact
     5161 ggactgattt cagtttacaa ctattctgtc tagtttaaga ctttattgtc atagtttaga
     5221 tctattttgt tcagtttaag actttattgt ccgcccacac ccgcttacgc agggcatcca
     5281 tttattactc aaccgtaacc gattttgcca ggttacgcgg ctggtcctct agaattccca
     5341 tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt tgagaggctc taagggcttc
     5401 tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag
     5461 ccttatatat tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg
     5521 atttatatta attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag
     5581 tacgttagcc atgagagctt agtacgttag ccatgagggt ttagttcgtt aaacatgaga
     5641 gcttagtacg ttaaacatga gagcttagta cgtgaaacat gagagcttag tacgtactat
     5701 caacaggttg aactgctgat cttcagatcc tctacgccgg acgcatcgtg gccggatcgg
     5761 gggggggggg cgctcggtct tgccttgctc gtcggtgatg tacttcacca gctccgcgaa
     5821 gtcgctcttc ttgatggagc gcatggggac gtgcttggca atcacgcgca ccccccggcc
     5881 gttttagcgg ctaaaaaagt catggctctg ccctcgggcg gaccacgccc atcatgacct
     5941 tgccaagctc gtcctgcttc tcttcgatct tcgccagcag ggcgaggatc gtggcatcac
     6001 cgaaccgcgc cgtgcgcggg tcgtcggtga gccagagttt cagcaggccg cccaggcggc
     6061 ccaggtcgcc attgatgcgg gccagctcgc ggacgtgctc atagtccacg acgcccgtga
     6121 ttttgtagcc ctggccgacg gccagcaggt aggccgacag gctcatgccg gccgccgccg
     6181 ccttttcctc aatcgctctt cgttcgtctg gaaggcagta caccttgata ggtgggctgc
     6241 ccttcctggt tggcttggtt tcatcagcca tccgcttgcc ctcatctgtt acgccggcgg
     6301 tagccggcca gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga
     6361 cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac
     6421 gccgttggat acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg
     6481 cgaaaaagga tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag
     6541 cggaaaagcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6601 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6661 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6721 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6781 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6841 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6901 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aa
//
