LOCUS       sequence-388755-        3720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP under the control of J23100 and RiboJ-UTR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 5 to 39)
  AUTHORS   Keating KW, Young EM
  TITLE     Systematic Part Transfer by Extending a Modular Toolkit to Diverse 
            Bacteria.
  JOURNAL   ACS Synth Biol. 2023 Jun 9. doi: 10.1021/acssynbio.3c00104.
  PUBMED    37294017
REFERENCE   2  (bases 1 to 3720)
  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"; journalName: "ACS Synth 
            Biol. 2023 Jun 9. doi: 10.1021/acssynbio.3c00104."
COMMENT     Sequence Label: pBBR1_DVK_AE(J23100-RiboJ-UTR1-E0040m-B0015)
FEATURES             Location/Qualifiers
     source          1..3720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..39
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     misc_RNA        46..95
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             125..147
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             157..870
                     /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(187..215)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     810..830
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      889..960
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      976..1003
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      1151..1922
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1923..2585
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     oriT            2596..2704
                     /note="incP origin of transfer"
     primer_bind     complement(2794..2813)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2805..3620
                     /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             2805..3620
                     /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(2878..2897)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ggagttgacg gctagctcag tcctaggtac agtgctagct actagctgtc accggatgtg
       61 ctttccggtc tgatgagtcc gtgaggacga aacagcctct acaaataatt ttgtttaaaa
      121 taattttgtt taactttaag aaggagatat acaatgatgc gtaaaggaga agaacttttc
      181 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
      241 gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc
      301 actactggaa aactacctgt tccatggcca acacttgtca ctactttcgg ttatggtgtt
      361 caatgctttg cgagataccc agatcatatg aaacagcatg actttttcaa gagtgccatg
      421 cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca
      481 cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt
      541 gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac
      601 aatgtataca tcatggcaga caaacaaaag aatggaatca aagttaactt caaaattaga
      661 cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt
      721 ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg
      781 aaagatccca acgaaaagag agatcacatg gtccttcttg agtttgtaac agctgctggg
      841 attacacatg gcatggatga actatacaaa taataaaggt ccaggcatca aataaaacga
      901 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
      961 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatag cttatgtctt
     1021 cagcagtagg acaaatccgc cgccctagac agctgggcgc gcccccccta cgggcttgct
     1081 ctccgggctt cgccctgcgc ggtcgctgcg ctcccttgcc agcccgtgga tatgtggacg
     1141 atggccgcga gcggccaccg gctggctcgc ttcgctcggc ccgtggacaa ccctgctgga
     1201 caagctgatg gacaggctgc gcctgcccac gagcttgacc acagggattg cccaccggct
     1261 acccagcctt cgaccacata cccaccggct ccaactgcgc ggcctgcggc cttgccccat
     1321 caattttttt aattttctct ggggaaaagc ctccggcctg cggcctgcgc gcttcgcttg
     1381 ccggttggac accaagtgga aggcgggtca aggctcgcgc agcgaccgcg cagcggcttg
     1441 gccttgacgc gcctggaacg acccaagcct atgcgagtgg gggcagtcga agggcgaagc
     1501 ccgcccgcct gccccccgag cctcacggcg gcgagtgcgg gggttccaag ggggcagcgc
     1561 caccttgggc aaggccgaag gccgcgcagt cgatcaacaa gccccggagg ggccactttt
     1621 tgccggaggg ggagccgcgc cgaaggcgtg ggggaacccc gcaggggtgc ccttctttgg
     1681 gcaccaaaga actagatata gggcgaaatg cgaaagactt aaaaatcaac aacttaaaaa
     1741 aggggggtac gcaacagctc attgcggcac cccccgcaat agctcattgc gtaggttaaa
     1801 gaaaatctgt aattgactgc cacttttacg caacgcataa ttgttgtcgc gctgccgaaa
     1861 agttgcagct gattgcgcat ggtgccgcaa ccgtgcggca cccctaccgc atggagataa
     1921 gcatggccac gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg
     1981 tgcaaacgga acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg
     2041 cggcaatgct gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca
     2101 gccagaaaac actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca
     2161 aggacttggt ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt
     2221 cggcctacgt ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt
     2281 cggtgttcag tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc
     2341 atggcgacct gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc
     2401 ccggcgagga gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct
     2461 tgaccgaaac ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc
     2521 cgtgttttct ggacgatggc gagccgttgg agccgccgac acgggtaacg ctgccgcgcc
     2581 ggtagggccg gcctacggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg
     2641 cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct
     2701 gcccttgtgt ctcaaaatct ctgatgtaca ttgcacaaga taaaaatata tcatcatgaa
     2761 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     2821 gggaaacgtc ttgctccagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     2881 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga
     2941 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     3001 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     3061 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag
     3121 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     3181 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     3241 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     3301 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt
     3361 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     3421 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     3481 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     3541 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     3601 tgctcgatga gtttttctaa ggtaactgtc agaccaagtt tactcatata tactttagat
     3661 tgatttttcg ttccactgag cgtcagaccc cagaactctg tcagaccgag tggaagacat
//
