LOCUS       sequence-420759-        3156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing green fluorescent protein under constitutive 
            promoter PgyrA control..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3156)
  TITLE     Harnessing xylitol utilization in Escherichia coli for 
            antibiotic-free plasmid and recombinant protein production.
REFERENCE   2  (bases 1 to 3156)
  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"
COMMENT     Sequence Label: PgyrA_gfp
FEATURES             Location/Qualifiers
     source          1..3156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     152..169
                     /label=L4440
                     /note="L4440 vector, forward primer"
     RBS             477..499
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(507..509,510..1223)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(507..509,510..1220)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(549..570)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(810..829)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1157..1178
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1584..2375
                     /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"
     rep_origin      2566..3154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3055..3074
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
       61 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      121 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      181 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatgctg
      241 gatccttccc tcgactcaca cttggggagg cgacggcttc aaatttagcg atctcttcgt
      301 ggtctacgtt atggttttcc ggcgattttt cggcattcat tggcacttct actccgtaat
      361 tggcaagaca aacgagtata tcaggcattg gatgtgaata aagcgtatag gtttacctca
      421 aactgcgcgg ctgtgttata atttgcgacc tttgaatccg ggatactaga aataattttg
      481 tttaacttta agaaggagat atacatatga gcaagggcga ggagctgttc accggggtgg
      541 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      601 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      661 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
      721 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      781 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
      841 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
      901 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
      961 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1021 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1081 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1141 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1201 gcatggacga gctgtacaag tgatgggctg aagggtttaa gcgaagacga acaataaggc
     1261 ctcccaaatc ggggggcctt ttttattgat aacaaaaagg caacactatg acatcggaaa
     1321 acccgttact ggcgctgcga gagaaaatca gcgcgctgga tgaaaaatta ttagcgttac
     1381 tggcagagcg gcgcgaattc aaacagaaag gccatgggtc gacctgcaga agcttagatc
     1441 tattaccctg ttatccctac tcgagttcat gtgcagctcc ataagcaaaa ggggatgata
     1501 agtttatcac caccgactat ttgcaacagt gccgttgatc gtgctatgat cgactgatgt
     1561 catcagcggt ggagtgcaat gtcatgaggg aagcggtgat cgccgaagta tcgactcaac
     1621 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt
     1681 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta
     1741 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa
     1801 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc
     1861 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc
     1921 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta
     1981 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac
     2041 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat
     2101 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca
     2161 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa
     2221 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg
     2281 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg
     2341 tgaaaggcga gatcaccaag gtagtcggca aataagatgc cgctcgccag tcgattggct
     2401 gagctcataa gttcctattc cgaagttccg cgaacgcgta aaggatctag gtgaagatcc
     2461 tttttgataa tctcatgacc aaaatccctt aacgtcacac aacatagcca cggagttttc
     2521 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2581 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2641 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2701 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2761 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2821 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2881 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2941 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3001 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3061 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3121 gtgatgctcg tcaggggggc ggagcctatg gaaaaa
//
