LOCUS       sequence-42989-e        3514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3514)
  AUTHORS   Lee TS, Krupa RA, Zhang F, Hajimorad M, Holtz WJ, Prasad N, Lee SK, 
            Keasling JD
  TITLE     BglBrick vectors and datasheets: A synthetic biology platform for 
            gene expression.
  JOURNAL   J Biol Eng. 2011 Sep 20;5:12.
  PUBMED    21933410
REFERENCE   2  (bases 1 to 3514)
  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: "J Biol 
            Eng."; date: "2011-09-20"; volume: "5"; pages: "12"
COMMENT     Sequence Label: pBbE2a-RFP
FEATURES             Location/Qualifiers
     source          1..3514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..633)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     538..557
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        649..704
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    685..703
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             742..1419
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1452..1523
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1539..1566
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1724..2312)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1804..1823)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2400..2494)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2524..3315,3316..3384))
                     /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     3147..3166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3385..3489)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc
       61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg
      121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg
      181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa
      241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc
      301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag
      361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc
      421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa
      481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc
      541 gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt
      601 aatcacttta cttttatcta atctagacat cattaattcc taatttttgt tgacactcta
      661 tcgttgatag agttatttta ccactcccta tcagtgatag agaaaagaat tcaaaagatc
      721 ttttaagaag gagatataca tatggcgagt agcgaagacg ttatcaaaga gttcatgcgt
      781 ttcaaagttc gtatggaagg ttccgttaac ggtcacgagt tcgaaatcga aggtgaaggt
      841 gaaggtcgtc cgtacgaagg tacccagacc gctaaactga aagttaccaa aggtggtccg
      901 ctgccgttcg cttgggacat cctgtccccg cagttccagt acggttccaa agcttacgtt
      961 aaacacccgg ctgacatccc ggactacctg aaactgtcct tcccggaagg tttcaaatgg
     1021 gaacgtgtta tgaacttcga agacggtggt gttgttaccg ttacccagga ctcctccctg
     1081 caagacggtg agttcatcta caaagttaaa ctgcgtggta ccaacttccc gtccgacggt
     1141 ccggttatgc agaaaaaaac catgggttgg gaagcttcca ccgaacgtat gtacccggaa
     1201 gacggtgctc tgaaaggtga aatcaaaatg cgtctgaaac tgaaagacgg tggtcactac
     1261 gacgctgaag ttaaaaccac ctacatggct aaaaaaccgg ttcagctgcc gggtgcttac
     1321 aaaaccgaca tcaaactgga catcacctcc cacaacgaag actacaccat cgttgaacag
     1381 tacgaacgtg ctgaaggtcg tcactccacc ggtgcttaag gatccaaact cgagtaagga
     1441 tctccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     1501 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt
     1561 tctgcgttta tacctagggc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1621 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1681 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1741 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1801 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1861 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1921 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1981 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2041 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2101 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2161 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2221 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2281 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2341 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgact agtgcttgga
     2401 ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt
     2461 tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgtaaa cttggtctga
     2521 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2581 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2641 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     2701 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2761 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     2821 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     2881 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     2941 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3001 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3061 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3121 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3181 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3241 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3301 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3361 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3421 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3481 ggttccgcgc acatttcccc gaaaagtgcc acct
//
