LOCUS       sequence-232203-        7499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial eGFP labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7499)
  AUTHORS   Kang YS, Kirby JE
  TITLE     Promotion and Rescue of Intracellular Brucella neotomae Replication 
            during Coinfection with Legionella pneumophila.
  JOURNAL   Infect Immun. 2017 Apr 21;85(5). pii: e00991-16. doi: 
            10.1128/IAI.00991-16. Print 2017 May.
  PUBMED    28264909
REFERENCE   2  (bases 1 to 7499)
  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: "Infect 
            Immun."; date: "2017-04-21"; volume: "85(5). pii: e00991-16. doi"; 
            pages: " 10.1128/IAI.00991-16. Print 2017 May"
COMMENT     Sequence Label: pAT28-proD/SaEGFP
FEATURES             Location/Qualifiers
     source          1..7499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    95..116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(131..136,137..154,155..161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    169..185
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     174..196
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     193..209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     193..209
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(386..388,389..391,392..1102)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1133..1150)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1133..1149)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1142..1164)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(1475..1846)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            1879..1988
                     /note="incP origin of transfer"
     rep_origin      6718..7306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7207..7226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7460..7477
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      181 aacaatttca cacaggaaac agctatgaca tgattacgaa ttccacagct aacaccacgt
      241 cgtccctatc tgctgcccta ggtctatgag tggttgctgg ataactttac gggcatgcat
      301 aaggctcgta taatatattc agggagacca caacggtttc cctctacaaa taattttgtt
      361 taactttgaa ttccaggagg actctatggt tagtaaagga gaagaattat ttacaggtgt
      421 agttccaata ttagttgaat tagatggtga cgttaatggt cacaaatttt cagtttcagg
      481 tgaaggtgaa ggtgatgcga cttatggaaa actaacatta aagtttattt gtacaactgg
      541 aaaattacct gtaccgtggc cgacattagt gactacactt acatatggtg tgcaatgctt
      601 ctcacgttat ccagatcata tgaagcaaca tgattttttt aaatctgcaa tgcctgaggg
      661 atatgttcaa gaaagaacta tcttttttaa agatgatgga aattataaaa ctcgtgcgga
      721 ggttaagttc gaaggtgaca cgttagtaaa tcgtatcgaa ttaaaaggta ttgatttcaa
      781 agaagacggc aatattttag gtcacaaatt agaatacaat tataattcac acaatgttta
      841 tattatggca gacaaacaaa aaaacggtat taaggtgaat tttaaaatac gtcataatat
      901 cgaagatggt tctgtgcaac tagcagatca ttatcaacag aatactccaa ttggtgatgg
      961 tccagtgtta ttacctgaca atcattattt aagtactcag tcagcattaa gtaaagaccc
     1021 aaatgagaag cgtgatcaca tggttttatt agaatttgtt acagcagcag gtattactct
     1081 tggtatggat gaattatata aataagtcga cctgcaggca tgcaagcttg gcactggccg
     1141 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     1201 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     1261 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     1321 tgtgcggtat ttcacaccgc ataggggatc ggtcttgcct tgctcgtcgg tgatgtactt
     1381 caccagctcc gcgaagtcgc tcttcttgat ggagcgcatg gggacgtgct tggcaatcac
     1441 gcgcaccccc cggccgtttt agcggctaaa aaagtcatgg ctctgccctc gggcggacca
     1501 cgcccatcat gaccttgcca agctcgtcct gcttctcttc gatcttcgcc agcagggcga
     1561 ggatcgtggc atcaccgaac cgcgccgtgc gcgggtcgtc ggtgagccag agtttcagca
     1621 ggccgcccag gcggcccagg tcgccattga tgcgggccag ctcgcggacg tgctcatagt
     1681 ccacgacgcc cgtgattttg tagccctggc cgacggccag caggtaggcc gacaggctca
     1741 tgccggccgc cgccgccttt tcctcaatcg ctcttcgttc gtctggaagg cagtacacct
     1801 tgataggtgg gctgcccttc ctggttggct tggtttcatc agccatccgc ttgccctcat
     1861 ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg attcccgttg agcaccgcca
     1921 ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct acttcaccta
     1981 tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc ctttggcaaa
     2041 atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata atgaccccga
     2101 agcagggtta tgcagcggaa aagatccggg ggatcgatcc tctagctaga ggatcgatct
     2161 gtataataaa gaataattat taatctgtag acaaattgtg aaaggatgta cttaaacgct
     2221 aacggtcagc tttattgaac agtaatttaa gtatatgtcc aatctagggt aagtaaattg
     2281 agtatcaata taaactttat atgaacataa tcaacgaggt gaaatcatga gcaatttgat
     2341 taacggaaaa ataccaaatc aagcgattca aacattaaaa atcgtaaaag atttatttgg
     2401 aagttcaata gttggagtat atctatttgg ttcagcagta aatggtggtt tacgcattaa
     2461 cagcgatgta gatgttctag tcgtcgtgaa tcatagttta cctcaattaa ctcgaaaaaa
     2521 actaacagaa agactaatga ctatatcagg aaagattgga aatacggatt ctgttagacc
     2581 acttgaagtt acggttataa ataggagtga agttgtccct tggcaatatc ctccaaaaag
     2641 agaatttata tacggtgagt ggctcagggg tgaatttgag aatggacaaa ttcaggaacc
     2701 aagctatgat cctgatttgg ctattgtttt agcacaagca agaaagaata gtatttctct
     2761 atttggtcct gattcttcaa gtatacttgt ctccgtacct ttgacagata ttcgaagagc
     2821 aattaaggat tctttgccag aactaattga ggggataaaa ggtgatgagc gtaatgtaat
     2881 tttaacccta gctcgaatgt ggcaaacagt gactactggt gaaattacct cgaaagatgt
     2941 cgctgcagaa tgggctatac ctcttttacc taaagagcat gtaactttac tggatatagc
     3001 tagaaaaggc tatcggggag agtgtgatga taagtgggaa ggactatatt caaaggtgaa
     3061 agcactcgtt aagtatatga aaaattctat agaaacttct ctcaattagg ctaattttat
     3121 tgcaataaca ggtgcttact tttaaaacta ctgatttatt gataaatatt gaacaatttt
     3181 tgggaagaat aaagcgtcct cttgtgaaat tagagaacgc tttattactt taatttagtg
     3241 aaacaatttg taactattga aaatagaaag aaattgttcc ttcgatagtt tattaatatt
     3301 agtgacattt cttcaaagcc tgtcggaatt ggtttttagt gaaaagaaca cgatattcac
     3361 ggtttaccca cttataaaca aaagatcgga tcgatcccca aatcaaaaga aaaattctcc
     3421 caagaattag aaatgagtag atcaaattat tcacgaatag aatcaggaaa atcagatcca
     3481 accataaaaa cactagaaca aattgcaaag ttaactaact caacgctagt agtggattta
     3541 atcccaaatg agccaacaga accagaacca gaaacagaat cagaacaagt aacattggat
     3601 ttagaaatgg aagaagaaaa aagcaatgac ttcgtgtgaa taatgcacga aatcgttgct
     3661 tatttttttt aaaagcggta tactagatat aacgaaacaa cgaactgaat agaaacgaaa
     3721 aaagagccat gacacattta taaaatgttt gacgacattt tataaatgca tagcccgata
     3781 agattgccaa accaacgctt atcagttagt cagatgaact cttccctcgt aagaagttat
     3841 ttaattaact ttgtttgaag acggtatata accgtactat cattatatag ggaaatcaga
     3901 gagttttcaa gtatctaagc tactgaattt aagaattgtt aagcaatcaa tcggaaatcg
     3961 tttgattgct ttttttgtat tcatttatag aaggtggagt ttgtatgaat catgatgaat
     4021 gtaaaactta tataaaaaat agtttattgg agataagaaa attagcaaat atctatacac
     4081 tagaaacgtt taagaaagag ttagaaaaga gaaatatcta cttagaaaca aaatcagata
     4141 agtatttttc ttcggagggg gaagattata tatataagtt aatagaaaat aacaaaataa
     4201 tttattcgat tagtggaaaa aaattgactt ataaaggaaa aaaatctttt tcaaaacatg
     4261 caatattgaa acagttgaat gaaaaagcaa accaagttaa ttaaacaacc tattttatag
     4321 gatttatagg aaaggagaac agctgaatga atatcccttt tgttgtagaa actgtgcttc
     4381 atgacggctt gttaaagtac aaatttaaaa atagtaaaat tcgctcaatc actaccaagc
     4441 caggtaaaag caaaggggct atttttgcgt atcgctcaaa atcaagcatg attggcggtc
     4501 gtggtgttgt tctgacttcc gaggaagcga ttcaagaaaa tcaagataca tttacacatt
     4561 ggacacccaa cgtttatcgt tatggaacgt atgcagacga aaaccgttca tacacgaaag
     4621 gacattctga aaacaattta agacaaatca ataccttctt tattgatttt gatattcaca
     4681 cggcaaaaga aactatttca gcaagcgata ttttaacaac cgctattgat ttaggtttta
     4741 tgcctactat gattatcaaa tctgataaag gttatcaagc atattttgtt ttagaaacgc
     4801 cagtctatgt gacttcaaaa tcagaattta aatctgtcaa agcagccaaa ataatttcgc
     4861 aaaatatccg agaatatttt ggaaagtctt tgccagttga tctaacgtgt aatcattttg
     4921 gtattgctcg cataccaaga acggacaatg tagaattttt tgatcctaat taccgttatt
     4981 ctttcaaaga atggcaagat tggtctttca aacaaacaga taataagggc tttactcgtt
     5041 caagtctaac ggttttaagc ggtacagaag gcaaaaaaca agtagatgaa ccctggttta
     5101 atctcttatt gcacgaaacg aaattttcag gagaaaaggg tttaataggg cgtaataacg
     5161 tcatgtttac cctctcttta gcctacttta gttcaggcta ttcaatcgaa acgtgcgaat
     5221 ataatatgtt tgagtttaat aatcgattag atcaaccctt agaagaaaaa gaagtaatca
     5281 aaattgttag aagtgcctat tcagaaaact atcaaggggc taatagggaa tacattacca
     5341 ttctttgcaa agcttgggta tcaagtgatt taaccagtaa agatttattt gtccgtcaag
     5401 ggtggtttaa attcaagaaa aaaagaagcg aacgtcaacg tgttcatttg tcagaatgga
     5461 aagaagattt aatggcttat attagcgaaa aaagcgatgt atacaagcct tatttagtga
     5521 cgaccaaaaa agagattaga gaagtgctag gcattcctga acggacatta gataaattgc
     5581 tgaaggtact gaaggcgaat caggaaattt tctttaagat taaaccagga agaaatggtg
     5641 gcattcaact tgctagtgtt aaatcattgt tgctatcgat cattaaagta aaaaaagaag
     5701 aaaaagaaag ctatataaag gcgctgacaa attcttttga cttagagcat acattcattc
     5761 aagagacttt aaacaagcta gcagaacgcc ctaaaacgga cacacaactc gatttgttta
     5821 gctatgatac aggctgaaaa taaaacccgc actatgccat tacatttata tctatgatac
     5881 gtgtttgttt tttctttgct gtttagcgaa tgattagcag aaatatacag agtaagattt
     5941 taattaatta ttagggggag aaggagagag tagcccgaaa acttttagtt ggcttggact
     6001 gaacgaagtg agggaaaggc tactaaaacg tcgaggggca gtgagagcga agcgaacact
     6061 tgatttttta attttctatc ttttataggt cattagagta tacttatttg tcctataaac
     6121 tatttagcag cataatagat ttattgaata ggtcatttaa gttgagcata ttagaggagg
     6181 aaaatcttgg agaaatattt gaagaacccg attacatgga ttggattagt tcttgtggtt
     6241 acgtggtttt taactaaaag tagtgaattt ttgatttttg gtgtgtgtgt cttgttgtta
     6301 gtatttgcta gtcaaagtga ttaaatagaa ttgaccactt ctgcgctcgg cccttccggc
     6361 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6421 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6481 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6541 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6601 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6661 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6721 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6781 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6841 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6901 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6961 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7021 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7081 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7141 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7201 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7261 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7321 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7381 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     7441 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatac
//
