LOCUS       sequence-342924-        4625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4625)
  AUTHORS   
  TITLE     Turn on the Mtr pathway genes under pLacI promoter in Shewanella 
            oneidensis MR-1
  JOURNAL   Bioresour. Bioprocess. 5, 35 (2018)
REFERENCE   2  (bases 1 to 4625)
  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: "Bioresour. 
            Bioprocess. 5, 35 (2018)"
COMMENT     Sequence Label: pETSXM2-pLacI-GFP
FEATURES             Location/Qualifiers
     source          1..4625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1154..1174
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      complement(1181..1208)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1224..1295)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1318..2031)
                     /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(1358..1378)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     1973..2001
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             2038..2049
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2067..2083
                     /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)."
     promoter        complement(join(2091..2097,2098..2115,2116..2121))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2136..2157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    2258..2279
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      2282..2325
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     complement(2329..2347)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      complement(2575..3163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2655..2674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3274..3293)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3285..4100
                     /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             3285..4100
                     /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(3358..3377)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(join(4192..4625,1..22))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4329..4350)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4541..4560
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 aatgcgccgc tacagggcgc gtcccattcg ccaatccgga tatagttcct cctttcagca
       61 aaaaacccct caagacccgt ttagaggccc caagggatcc gagcctttag gcgaactagg
      121 gcgaagccct tggtcgcggc gcagccgcca gatgctcaaa tttacttgtt tatccacagg
      181 cgaaaccctt tttaatagat ctttcttgtt ttttaagatc tttcttgttt ttagcgccgc
      241 tgtagccctt ggtgctaaag ggctacagcg gtttaatgtg gagagattta cggtaaaaca
      301 gcccctaaag gtggagagat ttacggtaaa acaatcccta aaggtggaga gatttacggt
      361 gatattcaca gttaattgtg gagaacgttg atagcaattc cacaattggt tattgtggaa
      421 tccccaaaaa taaactccac aatagtgacg taaaatgaac aacctatctg taactaaatc
      481 aaatagcttg atagatgcta gttacaaact aaatgtacaa gctcaaaaac tggtcttagc
      541 ttgcctcggg aaagtagatt ctagaggtaa tgcaccaaag gaaatgaccc taacagccct
      601 tgaattctca gaacttatgg ggatagacat caaaaatgct catagagagc tatacaaggc
      661 gtctgatacc cttttcgatg ccgtaatact cctaagagat gagcaagaag aggtaaagct
      721 taggtggatt caaaaaggag tgaaaaaact caaaggacaa ggagcaataa caatcacttg
      781 gactgatgaa gttttgaagt acatcagtag tttacagagc aggtttacaa cttataagct
      841 gcgacatatt gcgaatctac agtcagctca ttctattcgt ttatacgaac ttttgatgaa
      901 atttaacgct actggagaaa gagttatcta tgtggatgac ttccgctcag cattaggtat
      961 ctccgacaaa tacccacaat ttagagactt aaataaatgg gttatcaagc cggccgtaga
     1021 tgaactaaat cagcgttcag acctcaccat aaattacgag accattaaga aagggcgaac
     1081 agtggctgca ttgtcatttg agttcaagca gagcgcacag ctaaaattag acgtataatc
     1141 agtcgaccca tggctgcagc ggccgctact agtatataaa cgcagaaagg cccacccgaa
     1201 ggtgagccag tgtgactcta gtagagagcg ttcaccgaca aacaacagat aaaacgaaag
     1261 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggctctagt attattattt
     1321 gtatagttca tccatgccat gtgtaatccc agcagctgtt acaaactcaa gaaggaccat
     1381 gtggtctctc ttttcgttgg gatctttcga aagggcagat tgtgtggaca ggtaatggtt
     1441 gtctggtaaa aggacagggc catcgccaat tggagtattt tgttgataat ggtctgctag
     1501 ttgaacgctt ccatcttcaa tgttgtgtct aattttgaag ttaactttga ttccattctt
     1561 ttgtttgtct gccatgatgt atacattgtg tgagttatag ttgtattcca atttgtgtcc
     1621 aagaatgttt ccatcttctt taaaatcaat accttttaac tcgattctat taacaagggt
     1681 atcaccttca aacttgactt cagcacgtgt cttgtagttc ccgtcatctt tgaaaaatat
     1741 agttctttcc tgtacataac cttcgggcat ggcactcttg aaaaagtcat gctgtttcat
     1801 atgatctggg tatctcgcaa agcattgaac accataaccg aaagtagtga caagtgttgg
     1861 ccatggaaca ggtagttttc cagtagtgca aataaattta agggtaagtt ttccgtatgt
     1921 tgcatcacct tcaccctctc cactgacaga aaatttgtgc ccattaacat caccatctaa
     1981 ttcaacaaga attgggacaa ctccagtgaa aagttcttct cctttacgca tctagtattt
     2041 ctcctctttc tctagtatgt gtgaaattgt tatccgctca caattccaca caacatacga
     2101 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     2161 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2221 atcggccaac gcgcggggag aggcggtttg cgtattgctc tagaagcggc cgcgaattcc
     2281 agaaatcatc cttagcgaaa gctaaggatt ttttttatct gaaattctgc ctcgtgatac
     2341 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactc
     2401 gagaatgcaa cacacgcaac accgcatctc ttgatgacta gtgcgagcgg tatcagctca
     2461 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2521 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2581 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga agtggcgaaa
     2641 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2701 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2761 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2821 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2881 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2941 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3001 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3061 aaaaagagtt ggtagctctt gatccggcaa actaaccacc gctggtagcg gtggtttttt
     3121 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3181 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgaa
     3241 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     3301 gggaaacgtc ttgctctagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     3361 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga
     3421 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     3481 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     3541 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag
     3601 cattccaggt attagaagag tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     3661 tgttcctgcg ccggttgcac tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     3721 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     3781 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     3841 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     3901 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     3961 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     4021 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     4081 tgctcgatga gtttttctaa gaattaattc atgagcggat acatatttga atgtatttag
     4141 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     4201 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4261 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     4321 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4381 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     4441 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4501 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4561 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4621 cgctt
//
