LOCUS       sequence-203507-        3831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  R6 plasmid. Contains PBAD-EGFP and is part of part of the CAMERA 
            system for rewritable multi-event analog recording in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3831)
  AUTHORS   Tang W, Liu DR
  TITLE     Rewritable multi-event analog recording in bacterial and mammalian 
            cells.
  JOURNAL   Science. 2018 Feb 15. pii: science.aap8992. doi: 
            10.1126/science.aap8992.
  PUBMED    29449507
REFERENCE   2  (bases 1 to 3831)
  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: "Science. 
            2018 Feb 15. pii: science.aap8992. doi: 10.1126/science.aap8992."
COMMENT     Sequence Label: pWT009d
FEATURES             Location/Qualifiers
     source          1..3831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(17..808,809..877))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     640..659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(878..969)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(985..1734)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(995..1012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1235..1254)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      1823..1868
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1883..2761)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2788..3072
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2995..3014
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             join(3112..3114,3115..3117,3118..3831)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3112..3114,3115..3117,3118..3828)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3157..3178)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3418..3437)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3765..3786
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gacgaaaggg cctcgtttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
       61 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
      121 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
      181 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
      241 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
      301 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
      361 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
      421 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
      481 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
      541 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
      601 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
      661 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
      721 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
      781 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
      841 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
      901 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
      961 ataaacaaat aggcatgcag cgctcttccg cttcctcgct cactgactcg ctacgctcgg
     1021 tcgttcgact gcggcgagcg gtgtcagctc actcaaaagc ggtaatacgg ttatccacag
     1081 aatcagggga taaagccgga aagaacatgt gagcaaaaag caaagcaccg gaagaagcca
     1141 acgccgcagg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1201 tcaagccaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1261 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1321 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gttggtatct cagttcggtg
     1381 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1441 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1501 gcagcagcca ttggtaactg atttagagga ctttgtcttg aagttatgca cctgttaagg
     1561 ctaaactgaa agaacagatt ttggtgagtg cggtcctcca acccacttac cttggttcaa
     1621 agagttggta gctcagcgaa ccttgagaaa accaccgttg gtagcggtgg tttttcttta
     1681 tttatgagat gatgaatcaa tcggtctatc aagtcaacga acagctattc cgttactcta
     1741 gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa
     1801 gttgtaaaag aggacatccg gtcaaataaa acgaaaggct cagtcgaaag actgggcctt
     1861 tcgttttaga cttagggacc ctttatgaca acttgacggc tacatcattc actttttctt
     1921 cacaaccggc acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga
     1981 aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg
     2041 tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa
     2101 tccctaactg ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg
     2161 cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct
     2221 cgcgtacccg attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca
     2281 gtaacaattg ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc
     2341 cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc
     2401 gaaagaaccc cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc
     2461 gcggacgaaa gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt
     2521 gatgaatctc tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc
     2581 cctgattttt caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc
     2641 ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg
     2701 ccaccagatg ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca
     2761 tacttttcat actcccacca ttcagagaag aaaccaattg tccatattgc atcagacatt
     2821 gccgtcactg cgtcttttac tggctcttct cgctaaccca accggtaacc ccgcttatta
     2881 aaagcattct gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct
     2941 ataatcacgg cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca
     3001 tagcattttt atccataaga ttagcggatc ctacctgacg ctttttatcg caactctcta
     3061 ctgtttctcc atacccgttt tacctgcagg tgcagtaagg aggaaaaaaa aatggtgagc
     3121 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     3181 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     3241 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     3301 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     3361 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     3421 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     3481 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     3541 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     3601 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     3661 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     3721 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     3781 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta a
//
