LOCUS       sequence-83230-e        6068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Controls GFP expression from the P3 promoter cloned out of S. aureus
            RN4220 genome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6068)
  AUTHORS   Marchand N, Collins CH
  TITLE     Peptide-based communication system enables Escherichia coli to 
            Bacillus megaterium interspecies signaling.
  JOURNAL   Biotechnol Bioeng. 2013 Nov;110(11):3003-12. doi: 10.1002/bit.24975.
            Epub 2013 Jul 5.
  PUBMED    23775238
REFERENCE   2  (bases 1 to 6068)
  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"; doi: "10.1002/bit.24975";
            journalName: "Biotechnol Bioeng"; date: "2013-11"; volume: "110"; 
            issue: "11"; pages: "3003-12"
COMMENT     Sequence Label: pP3-GFP
FEATURES             Location/Qualifiers
     source          1..6068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(661..663,664..1377)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="ASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDEPYK"
     primer_bind     complement(691..719)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1314..1334
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1418..1465
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1621..2209)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1701..1720)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2380..3171,3172..3240))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3003..3022
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3241..3345)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3413..3431
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(3509..4885)
                     /codon_start=1
                     /gene="Bacillus subtilis tetL"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     CDS             complement(join(5085..6068,1..21))
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
ORIGIN
        1 aatattaaaa gaaaccccca tctatttagt tatttgtttg gtcacttata actttaacag
       61 atggggtttt tctgtgcaac caattttaag ggttttcaat actttaaaac acatacatac
      121 caacacttca acgcaccttt cagcaactaa aataaaaatg acgttatttc tatatgtatc
      181 aagataagaa agaacaagtt caaaaccatc aaaaaaagac accttttcag gtgctttttt
      241 tattttataa actcattccc tgatctcgac ttcgttcttt ttttacctct cggttatgag
      301 ttagttcaaa ttcgttcttt ttaggttcta aatcgtgttt ttcttggaat tgtgctgttt
      361 tatcctttac cttgtctaca aaccccttaa aaacgttttt aaaggctttt aagccgtctg
      421 tacgttcctt aaggcctaac tgtaggaaat aaatacttaa ctgttaaatg taatttgtat
      481 ttaatatttt aacataaaaa aatttacagt taagaataaa aaacgactag ttaagaaaaa
      541 ttggaaaata aatgctttta gcatgtttta atataactag atcacagaga tgtggtcagt
      601 cagtcttaat taaccaagga ggtgaaatgt acaatggtcc aaactagttc gaagatctgg
      661 gctagcaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
      721 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc tacatacgga
      781 aagcttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
      841 gtcactactt tgacctatgg tgttcaatgc ttttcccgtt atccggatca tatgaaacgg
      901 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaacgcac tatatctttc
      961 aaagatgacg ggaactacaa gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt
     1021 aatcgtatcg agttaaaagg tattgatttt aaagaagatg gaaacattct cggacacaaa
     1081 ctcgagtaca actataactc acacaatgta tacatcacgg cagacaaaca aaagaatgga
     1141 atcaaagcta acttcaaaat tcgccacaac attgaagatg gatccgttca actagcagac
     1201 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
     1261 ctgtcgacac aatctgccct ttcgaaagat cccaacgaaa agcgtgacca catggtcctt
     1321 cttgagtttg taactgctgc tgggattaca catggcatgg atgagcccta caaataatga
     1381 attcgcggcc gcatgccggc taatcgcgac cggttaacta gcataacccc ttggggcctc
     1441 taaacgggtc ttgaggggtt ttttgctaaa ggaggaacta tatccggtcc aagaattgga
     1501 gccaatcaat tcttgcggag aactgtgaat gcgcaaacca acccttggca gaacatatcc
     1561 atcgcgtccg ccatctccag cagccgcacg cggcgcatct cgggccgcgt tgctggcgtt
     1621 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1681 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1741 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1801 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1861 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1921 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1981 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2041 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     2101 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2161 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2221 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2281 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2341 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2401 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2461 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2521 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2581 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2641 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg
     2701 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2761 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     2821 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     2881 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     2941 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg
     3001 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3061 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3121 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3181 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3241 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3301 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3361 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     3421 tatcacgagg ccctttcgtc ttcaagaatt cctgttataa aaaaaggatc aattttgaac
     3481 tctctcccaa agttgatccc ttaacgattt agaaatccct ttgagaatgt ttatatacat
     3541 tcaaggtaac cagccaacta atgacaatga ttcctgaaaa aagtaataac aaattactat
     3601 acagataagt tgactgatca acttccatag gtaacaacct ttgatcaagt aagggtatgg
     3661 ataataaacc acctacaatt gcaatacctg ttccctctga taaaaagctg gtaaagttaa
     3721 gcaaactcat tccagcacca gcttcctgct gtttcaagct acttgaaaca attgttgata
     3781 taactgtttt ggtgaacgaa agcccaccta aaacaaatac gattataatt gtcatgaacc
     3841 atgatgttgt ttctaaaaga aaggaagcag ttaaaaagct aacagaaaga aatgtaactc
     3901 cgatgtttaa cacgtataaa ggacctcttc tatcaacaag tatcccacca atgtagccga
     3961 aaataatgac actcattgtt ccagggaaaa taattacact tccgatttcg gcagtactta
     4021 gctggtgaac atctttcatc atataaggaa ccatagagac aaaccctgct actgttccaa
     4081 atataattcc cccacaaaga actccaatca taaaaggtat atttttccct aatccgggat
     4141 caacaaaagg atctgttact ttcctgatat gttttacaaa tatcaggaat gacagcacgc
     4201 taacgataag aaaagaaatg ctatatgatg ttgtaaacaa cataaaaaat acaatgccta
     4261 cagacattag tataattcct ttgatatcaa aatgaccttt tatccttact tctttcttta
     4321 ataatttcat aagaaacgga acagtgataa ttgttatcat aggaatgagt agaagatagg
     4381 accaatgaat ataatgggct atcattccac caatcgctgg accgactcct tctcccatgg
     4441 ctactatcga tccaataaga ccaaatgctt tacccctatt ttcctttgga atatagcgcg
     4501 caactacaac cattacgagt gctggaaatg cagctgcacc agccccttga ataaaacgag
     4561 ccataataag taaggaaaag aaagaatggc caacaaaccc aattaccgac ccgaaacaat
     4621 ttattataat tccaaatagg agtaaccttt tgatgcctaa ttgatcagat agctttccat
     4681 atacagctgt tccaatggaa aaggttaaca taaaggctgt gttcacccag tttgtactcg
     4741 caggtggttt attaaaatca tttgcaatat caggtaatga gacgttcaaa accatttcat
     4801 ttaatacgct aaaaaaagat aaaatgcaaa gccaaattaa aatttggttg tgtcgtaaat
     4861 tcgattgtga ataggatgta ttcacatttc accctccaat aatgagggca gacgtagttt
     4921 atagggttaa tgatacgctt ccctctttta attgaaccct gttacattca ttacacttca
     4981 taattaattc ctcctaaact tgattaaaac attttaccac atataaacta agttttaaat
     5041 tcagtatttc atcacttata caacaatatg gcccgtttgt tgaactactc tttaataaaa
     5101 taatttttcc gttcccaatt ccacattgca ataatagaaa atccatcttc atcggctttt
     5161 tcgtcatcat ctgtatgaat caaatcgcct tcttctgtgt catcaaggtt taatttttta
     5221 tgtatttctt ttaacaaacc accataggag attaaccttt tacggtgtaa accttcctcc
     5281 aaatcagaca aacgtttcaa attcttttct tcatcatcgg tcataaaatc cgtatccttt
     5341 acaggatatt ttgcagtttc gtcaattgcc gattgtatat ccgatttata tttatttttc
     5401 ggtcgaatca tttgaacttt tacatttgga tcatagtcta atttcattgc ctttttccaa
     5461 aattgaatcc attgtttttg attcacgtag ttttctgtat tcttaaaata agttggttcc
     5521 acacatacca atacatgcat gtgctgatta taagaattat ctttattatt tattgtcact
     5581 tccgttgcac gcataaaacc aacaagattt ttattaattt ttttatattg catcattcgg
     5641 cgaaatcctt gagccatatc tgacaaactc ttatttaatt cttcgccatc ataaacattt
     5701 ttaactgtta atgtgagaaa caaccaacga actgttggct tttgtttaat aacttcagca
     5761 acaacctttt gtgactgaat gccatgtttc attgctctcc tccagttgca cattggacaa
     5821 agcctggatt tacaaaacca cactcgatac aactttcttt cgcctgtttc acgattttgt
     5881 ttatactcta atatttcagc acaatctttt actctttcag cctttttaaa ttcaagaata
     5941 tgcagaagtt caaagtaatc aacattagcg attttctttt ctctccatgg tctcactttt
     6001 ccactttttg tcttgtccac taaaaccctt gatttttcat ctgaataaat gctactatta
     6061 ggacacat
//
