LOCUS       sequence-42997-e        4864 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 4864)
  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 4864)
  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: pBbS2k-RFP
FEATURES             Location/Qualifiers
     source          1..4864
                     /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"
     CDS             complement(2097..3047)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(3809..3903)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3934..4728)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4045..4064)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4655..4674
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
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 tacctagggt acgggttttg ctgcccgcaa acgggctgtt ctggtgttgc
     1621 tagtttgtta tcagaatcgc agatccggct tcagccggtt tgccggctga aagcgctatt
     1681 tcttccagaa ttgccatgat tttttcccca cgggaggcgt cactggctcc cgtgttgtcg
     1741 gcagctttga ttcgataagc agcatcgcct gtttcaggct gtctatgtgt gactgttgag
     1801 ctgtaacaag ttgtctcagg tgttcaattt catgttctag ttgctttgtt ttactggttt
     1861 cacctgttct attaggtgtt acatgctgtt catctgttac attgtcgatc tgttcatggt
     1921 gaacagcttt gaatgcacca aaaactcgta aaagctctga tgtatctatc ttttttacac
     1981 cgttttcatc tgtgcatatg gacagttttc cctttgatat gtaacggtga acagttgttc
     2041 tacttttgtt tgttagtctt gatgcttcac tgatagatac aagagccata agaacctcag
     2101 atccttccgt atttagccag tatgttctct agtgtggttc gttgtttttg cgtgagccat
     2161 gagaacgaac cattgagatc atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa
     2221 ctggtgagct gaatttttgc agttaaagca tcgtgtagtg tttttcttag tccgttatgt
     2281 aggtaggaat ctgatgtaat ggttgttggt attttgtcac cattcatttt tatctggttg
     2341 ttctcaagtt cggttacgag atccatttgt ctatctagtt caacttggaa aatcaacgta
     2401 tcagtcgggc ggcctcgctt atcaaccacc aatttcatat tgctgtaagt gtttaaatct
     2461 ttacttattg gtttcaaaac ccattggtta agccttttaa actcatggta gttattttca
     2521 agcattaaca tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt gtgagttttc
     2581 ttttgtgtta gttcttttaa taaccactca taaatcctca tagagtattt gttttcaaaa
     2641 gacttaacat gttccagatt atattttatg aattttttta actggaaaag ataaggcaat
     2701 atctcttcac taaaaactaa ttctaatttt tcgcttgaga acttggcata gtttgtccac
     2761 tggaaaatct caaagccttt aaccaaagga ttcctgattt ccacagttct cgtcatcagc
     2821 tctctggttg ctttagctaa tacaccataa gcattttccc tactgatgtt catcatctga
     2881 gcgtattggt tataagtgaa cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg
     2941 gggttgagta gtgccacaca gcataaaatt agcttggttt catgctccgt taagtcatag
     3001 cgactaatcg ctagttcatt tgctttgaaa acaactaatt cagacataca tctcaattgg
     3061 tctaggtgat tttaatcact ataccaattg agatgggcta gtcaatgata attactagtc
     3121 cttttcccgg gtgatctggg tatctgtaaa ttctgctaga cctttgctgg aaaacttgta
     3181 aattctgcta gaccctctgt aaattccgct agacctttgt gtgttttttt tgtttatatt
     3241 caagtggtta taatttatag aataaagaaa gaataaaaaa agataaaaag aatagatccc
     3301 agccctgtgt ataactcact actttagtca gttccgcagt attacaaaag gatgtcgcaa
     3361 acgctgtttg ctcctctaca aaacagacct taaaacccta aaggcttaag tagcaccctc
     3421 gcaagctcgg gcaaatcgct gaatattcct tttgtctccg accatcaggc acctgagtcg
     3481 ctgtcttttt cgtgacattc agttcgctgc gctcacggct ctggcagtga atgggggtaa
     3541 atggcactac aggcgccttt tatggattca tgcaaggaaa ctacccataa tacaagaaaa
     3601 gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg ctatgtggtg ctatctgact
     3661 ttttgctgtt cagcagttcc tgccctctga ttttccagtc tgaccacttc ggattatccc
     3721 gtgacaggtc attcagactg gctaatgcac ccagtaaggc agcggtatca tcaacaggct
     3781 tacccgtctt actgtcccta gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac
     3841 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
     3901 gtccaagcga gctctcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata
     3961 gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc
     4021 ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg
     4081 gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat
     4141 gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg
     4201 cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc
     4261 atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc
     4321 ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc
     4381 catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac
     4441 ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca
     4501 aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag
     4561 ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa
     4621 cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc
     4681 cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc
     4741 tcatcctgtc tcttgatcag atcatgatcc cctgcgccat cagatccttg gcggcaagaa
     4801 agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa
     4861 ttcc
//
