LOCUS       sequence-45102-e        5103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mycobacteria replicating vector expressing eGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5103)
  AUTHORS   Seeliger JC, Topp S, Sogi KM, Previti ML, Gallivan JP, Bertozzi CR
  TITLE     A riboswitch-based inducible gene expression system for 
            mycobacteria.
  JOURNAL   PLoS One. 2012;7(1):e29266. Epub 2012 Jan 18.
  PUBMED    22279533
REFERENCE   2  (bases 1 to 5103)
  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: "PLoS One.";
            date: "2012"; volume: "7(1)"; pages: "e29266. Epub 2012 Jan 18"
COMMENT     Sequence Label: pST5552
FEATURES             Location/Qualifiers
     source          1..5103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(2233..2235,2236..2949)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2233..2235,2236..2946)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2275..2296)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2536..2555)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2883..2904
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3002..3048
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3198..3217)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3209..4024
                     /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
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3282..3301)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4355..4943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4844..4863
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagcccacca gctccgtaag ttcgggcgct gtgtggctcg tacccgcgca ttcaggcggc
       61 agggggtcta acgggtctaa ggcggcgtgt acggccgcca cagcggctct cagcggcccg
      121 gaaacgtcct cgaaacgacg catgtgttcc tcctggttgg tacaggtggt tgggggtgct
      181 cggctgtcgc tggtgttcca ccaccagggc tcgacgggag agcgggggag tgtgcagttg
      241 tggggtggcc cctcagcgaa atatctgact tggagctcgt gtcggaccat acaccggtga
      301 ttaatcgtgg tctactacca agcgtgagcc acgtcgccga cgaatttgag cagctctggc
      361 tgccgtactg gccgctggca agcgacgatc tgctcgaggg gatctaccgc caaagccgcg
      421 cgtcggccct aggccgccgg tacatcgagg cgaacccaac agcgctggca aacctgctgg
      481 tcgtggacgt agaccatcca gacgcagcgc tccgagcgct cagcgcccgg gggtcccatc
      541 cgctgcccaa cgcgatcgtg ggcaatcgcg ccaacggcca cgcacacgca gtgtgggcac
      601 tcaacgcccc tgttccacgc accgaatacg cgcggcgtaa gccgctcgca tacatggcgg
      661 cgtgcgccga aggccttcgg cgcgccgtcg atggcgaccg cagttactca ggcctcatga
      721 ccaaaaaccc cggccacatc gcctgggaaa cggaatggct ccactcagat ctctacacac
      781 tcagccacat cgaggccgag ctcggcgcga acatgccacc gccgcgctgg cgtcagcaga
      841 ccacgtacaa agcggctccg acgccgctag ggcggaattg cgcactgttc gattccgtca
      901 ggttgtgggc ctatcgtccc gccctcatgc ggatctacct gccgacccgg aacgtggacg
      961 gactcggccg cgcgatctat gccgagtgcc acgcgcgaaa cgccgaattt ccgtgcaacg
     1021 acgtgtgtcc cggaccgcta ccggacagcg aggtccgcgc catcgccaac agcatttggc
     1081 gttggatcac aaccaagtcg cgcatttggg cggacgggat cgtggtctac gaggccacac
     1141 tcagtgcgcg ccagtcggcc atctcgcgga agggcgcagc agcgcgcacg gcggcgagca
     1201 cagttgcgcg gcgcgcaaag tccgcgtcag ccatggaggc attgctatga gcgacggcta
     1261 cagcgacggc tacaaccggc agccgactgt ccgcaaaaag cggcgcgtga ccgccgccga
     1321 aggcgctcga atcaccggac tatccgaacg ccacgtcgtc cggctcgtgg cgcaggaacg
     1381 cagcgagtgg ctcgccgagc aggctgcacg ccgcgaacgc atccgcgcct atcacgacga
     1441 cgagggccac tcttggccgc aaacggccaa acatttcggg ctgcatctgg acaccgttaa
     1501 gcgactcggc tatcgggcga ggaaagagcg tgcggcagaa caggaagcgg ctcaaaaggc
     1561 ccacaacgaa gccgacaatc caccgctgtt ctaacgcaat tggggagcgg gtgtcgcggg
     1621 ggttccgtgg ggggttccgt tgcaacgggt cggacaggta aaagtcctgg tagacgctag
     1681 ttttctggtt tgggccatgc ctgtctcgtt gcgtgtttcg ttgcgcccgt tttgaatacc
     1741 agccagacga gacggggttc tacgaatctt ggtcgatacc aagccatttc cgctgaatat
     1801 cggggagctc accgccagaa tcggtggttg tggtgatgta cgtggcgaac tccgttgtag
     1861 tgcctgtggt ggcatccgtg gcgcggccgc ggtaccagat ctttaaatct agaggtgacc
     1921 acaacgcgcc cgctttgatc ggggacgtct gcggccgacc atttacgggt cttgttgtcg
     1981 ttggcggtca tgggccgaac atactcaccc ggatcggagg gccgaggaca aggtcgaacg
     2041 aggggcatga cccggtgcgg ggcttcttgc actcggcata ggcgagtgct aagaataacg
     2101 ttggcactcg cgaccgggat acgactcact ataggtaccg gtgataccag catcgtcttg
     2161 atgcccttgg cagcaccctg ctaaggaggc aacaagatgg ccaagacaat tgcggatcca
     2221 gctgcagaat tcgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2281 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2341 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     2401 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     2461 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2521 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2581 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2641 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     2701 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     2761 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2821 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2881 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2941 tacaagtaaa agcttatcga tgtcgacgta gttaactagc gtacgatcga ctgccaggca
     3001 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatgc catcatggcc
     3061 gcggtgatca gctagccacc tgacgtcggg ggggggggaa agccacgttg tgtctcaaaa
     3121 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     3181 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc
     3241 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     3301 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccc atgcgccaga
     3361 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     3421 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     3481 tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga
     3541 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     3601 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3661 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3721 tggctggcct gttgaacaag tctggaaaga aatgcataat cttttgccat tctcaccgga
     3781 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3841 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3901 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3961 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     4021 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     4081 cggcggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat cacgcatctt
     4141 cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa ctggtccacc
     4201 tacaacaaag ctctcatcaa ccgtggctcc ctcactttct ggctggatga tggggcgatt
     4261 caggcctggt atgagtcagc aacaccttct tcacgaggca gacctcacta gttccatgag
     4321 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4381 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4441 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4501 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4561 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4621 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4681 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4741 gtgagctatg agaaagcgcc acgcttcccg aggggagaaa ggcggacagg tatccggtaa
     4801 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4861 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4921 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4981 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5041 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcaacg
     5101 cgt
//
