LOCUS       sequence-146408-        2919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GFP under the sulA promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2919)
  AUTHORS   Cui L, Bikard D
  TITLE     Consequences of Cas9 cleavage in the chromosome of Escherichia coli.
  JOURNAL   Nucleic Acids Res. 2016 Apr 8. pii: gkw223.
  PUBMED    27060147
REFERENCE   2  (bases 1 to 2919)
  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: "Nucleic 
            Acids Res. 2016 Apr 8. pii: gkw223."
COMMENT     Sequence Label: pZA31-sulA-GFP
FEATURES             Location/Qualifiers
     source          1..2919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     34..52
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             349..1062
                     /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="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMILLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(379..407)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     terminator      1095..1181
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1217..1234)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1351..1896)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1431..1450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2010..2104)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2125..2784)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2699..2718
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2785..2887)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccgatcggt
       61 attcaattgt gcccaacgtt gccaggtatt cttgtgattt atagatccgc ttataggaga
      121 ggcttttcat aaaattcctt ttaaaatcat aacataaaag aatgattcac attaacggat
      181 ccgttaacta cgaaaatagg caacttattc ttaaggggca agattaattt atgttttccc
      241 gtcaccaacg acaaaatttg cgaggctctt tccgaaaata gggttgatct ttgttgtcac
      301 tggatgtact gtacatccat acagtaactc acaggggctg gattgattat gagtaaagga
      361 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
      421 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
      481 aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt cactactttc
      541 gcgtatggtc ttcaatgctt tgcgagatac ccagatcata tgaaacagca tgactttttc
      601 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
      661 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag
      721 ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt ggaatacaac
      781 tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat caaagttaac
      841 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
      901 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
      961 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tgatccttct tgagtttgta
     1021 acagctgctg ggattacaca tggcatggat gaactataca aataaggatc ccatggtacg
     1081 cgtgctagag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     1141 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgccc tagacctagg
     1201 ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
     1261 cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca
     1321 gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca
     1381 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca
     1441 ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
     1501 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
     1561 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
     1621 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
     1681 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
     1741 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
     1801 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
     1861 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata
     1921 aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc
     1981 atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
     2041 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
     2101 agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg
     2161 taattcatta agcattctgc cgacatggaa gccatcacag acggcatgat gaacctgaat
     2221 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     2281 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     2341 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     2401 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     2461 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     2521 aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaatt ccggatgagc
     2581 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     2641 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     2701 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     2761 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
     2821 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     2881 ccgatcaacg tctcattttc gccagatatc gacgtctaa
//
