LOCUS       sequence-49429-e        5248 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 5248)
  AUTHORS   Rodrigo G, Landrain TE, Jaramillo A
  TITLE     De novo automated design of small RNA circuits for engineering 
            synthetic riboregulation in living cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Sep 18;109(38):15271-6. Epub 2012 Sep
            4.
  PUBMED    22949707
REFERENCE   2  (bases 1 to 5248)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2012-09-18"; volume: "109(38)"; pages: 
            "15271-6. Epub 2012 Sep 4"
COMMENT     Sequence Label: pRAJ11-12
FEATURES             Location/Qualifiers
     source          1..5248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(26..703)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             710..721
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(747..774)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(790..861)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    957..975
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    982..1000
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1045..1061
                     /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)."
     RBS             1108..1119
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1135..1845
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     misc_feature    1852..1872
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1908..1939
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2038..2853)
                     /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"
     primer_bind     2761..2780
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2845..2864
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(3211..4161)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4209..4431
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4923..5017)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(5173..5202)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agttattaag caccggtgga gtgacgacct tcagcacgtt
       61 cgtactgttc aacgatggtg tagtcttcgt tgtgggaggt gatgtccagt ttgatgtcgg
      121 ttttgtaagc acccggcagc tgaaccggtt ttttagccat gtaggtggtt ttaacttcag
      181 cgtcgtagtg accaccgtct ttcagtttca gacgcatttt gatttcacct ttcagagcac
      241 cgtcttccgg gtacatacgt tcggtggaag cttcccaacc catggttttt ttctgcataa
      301 ccggaccgtc ggacgggaag ttggtaccac gcagtttaac tttgtagatg aactcaccgt
      361 cttgcaggga ggagtcctgg gtaacggtaa caacaccacc gtcttcgaag ttcataacac
      421 gttcccattt gaaaccttcc gggaaggaca gtttcaggta gtccgggatg tcagccgggt
      481 gtttaacgta agctttggaa ccgtactgga actgcgggga caggatgtcc caagcgaacg
      541 gcagcggacc acctttggta actttcagtt tagcggtctg ggtaccttcg tacggacgac
      601 cttcaccttc accttcgatt tcgaactcgt gaccgttaac ggaaccttcc atacgaactt
      661 tgaaacgcat gaactctttg ataacgtctt cggaggaagc catctagtat ttctcctctt
      721 tctctagtgg atccgctagc tataaacgca gaaaggccca cccgaaggtg agccagtgtg
      781 actctagtag agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga
      841 ctgagccttt cgttttattt gatgcctggc ctagggcctc gcataaatct gtcacagcat
      901 caacgtttcc gctgaactgt gacagactca cacaatcaac cctcccgtgc tcagtatctc
      961 tatcactgat agggatgtca atctctatca ctgataggga atgcatgcta ggatgcatat
     1021 aaatgtgagc ggataacatt gacattgtga gcggataaca agatactgag cacacccagt
     1081 atcattctct tcttcctgcc cacgcggaaa gaggagaaag gtgtaatgac tagccgtaaa
     1141 ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg tgatgtcaac
     1201 ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg taaactgacg
     1261 ctgaagttca tctgtactac tggtaaactg ccggtacctt ggccgactct ggtaacgacg
     1321 ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca gcatgacttc
     1381 ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt taaggatgac
     1441 ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt aaaccgcatt
     1501 gagctgaaag gcattgactt taaagaagac ggcaatatcc tgggccataa gctggaatac
     1561 aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg cattaaagcg
     1621 aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga tcactaccag
     1681 caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta tctgagcacg
     1741 caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct gctggagttc
     1801 gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaatgatg atactagtag
     1861 cggccgctgc aggagtcact aagggttagt tagttagatt agcagaaagt caaaagcctc
     1921 cgaccggagg cttttgacta aaacttccct tggggttatc attggggctc actcaaaggc
     1981 ggtaatcaga taaaaaaaat ccttagcttt cgctaaggat gatttctgct agtattatta
     2041 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     2101 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccaaag
     2161 aatggcaagg tcctggtaac ggtctgcgat tccgacccgt ccaacatcaa tacaacctat
     2221 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     2281 atccggtgag aatggcaaga gcttgtgcat ttctttccag acttgttcaa caggccagcc
     2341 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcatgc gtgattgcgc
     2401 ctgagcaaga cgaaatacac gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     2461 taaccggcgc aggaacacgg ccagcgcatc aacaatattt tcacctgaat caggatattc
     2521 ttctaatacc tggaaggctg ttttcccagg aatcgcggtg gtgagtaacc acgcatcatc
     2581 aggagtacgg ataaaatgct tgatggtcgg gagaggcata aactccgtca gccagttgag
     2641 acggaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     2701 ctctggcgca tcgggcttcc catacaagcg atagattgtc gcacctgatt gcccgacatt
     2761 atcgcgagcc catttatacc catataaatc agcgtccatg ttggagttta agcgcggacg
     2821 ggagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     2881 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     2941 attttgagac acaacgtggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca
     3001 gctctagtag ttacattgtc gatctgttca tggtgaacag ctttaaatgc accaaaaact
     3061 cgtaaaagct ctgatgtatc tatctttttt acaccgtttt catctgtgca tatggacagt
     3121 tttccctttg atatctaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct
     3181 tcactgatag atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt
     3241 ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatgctt
     3301 actttgcatg tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa
     3361 agcatcgtgt agtgtttttc ttagtccgtt acgtaggtag gaatctgatg taatggttgt
     3421 tggtattttg tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat
     3481 ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac
     3541 caccaatttc atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg
     3601 gttaagcctt ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc
     3661 aaggctaatc tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca
     3721 ctcataaatc ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt
     3781 tatgaatttt tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa
     3841 tttttcgctt gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa
     3901 aggattcctg atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc
     3961 ataagcattt tccctactga tgttcatcat ctgagcgtat tggttataag tgaacgatac
     4021 cgtccgttct ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa
     4081 aattagcttg gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt
     4141 gaaaacaact aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca
     4201 attgagatgg gctagtcaat gataattaca tgtccttttc ctttgagttg tgggtatctg
     4261 taaattctgc tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc
     4321 cgctagacct ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa
     4381 gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta
     4441 gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag
     4501 accttaaaac cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat
     4561 tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg
     4621 ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga
     4681 ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt
     4741 ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct
     4801 ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat
     4861 gcacccagta aggcagcggt atcatcaaca ggcttacccg tcttactgtc cctagtgctt
     4921 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     4981 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcgt aaacttggtc
     5041 tgacagctct agctccggca aaaaaacggg caaggtgtca ccaccctgcc ctttttcttt
     5101 aaaaccgaaa agattacttc gcgtttgcca cctgacgtct aagaaaagga atattcagca
     5161 atttgcccgt gccgaagaaa ggcccacccg tgaaggtgag ccagtgagtt gattgctacg
     5221 taattagtta gttagccctt agtgactc
//
