LOCUS       sequence-252166-        3273 bp    DNA     circular SYN 14-APR-2025
DEFINITION  For course in ""Synthetic Biology: A Lab Manual"".
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3273)
  AUTHORS   Liljeruhm J, Funk SK, Tietscher S, Edlund AD, Jamal S, Wistrand-Yuen
            P, Dyrhage K, Gynna A, Ivermark K, Lovgren J, Tornblom V, Virtanen 
            A, Lundin ER, Wistrand-Yuen E, Forster AC
  TITLE     Engineering a palette of eukaryotic chromoproteins for bacterial 
            synthetic biology.
  JOURNAL   J Biol Eng. 2018 May 10;12:8. doi: 10.1186/s13036-018-0100-0. 
            eCollection 2018.
  PUBMED    29760772
REFERENCE   2  (bases 1 to 3273)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Eng."; date: "2018-05-10"; volume: "12:8. doi"; pages: " 
            10.1186/s13036-018-0100-0. eCollection 2018"
COMMENT     Sequence Label: pSB1K3 containing mRFP1
FEATURES             Location/Qualifiers
     source          1..3273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(15..20,21..38,39..45)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    53..69
                     /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             87..98
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             105..782
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      827..898
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      914..941
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    948..968
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      969..1026
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1050..1067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1221..1809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1301..1320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2042..2857)
                     /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"
     CDS             complement(2042..2857)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /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     2765..2784
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2849..2868
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3062..3080
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3084..3127)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3130..3151
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    3252..3273
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
       61 ggataacaat ttcacacata ctagagaaag aggagaaata ctagatggct tcctccgaag
      121 acgttatcaa agagttcatg cgtttcaaag ttcgtatgga aggttccgtt aacggtcacg
      181 agttcgaaat cgaaggtgaa ggtgaaggtc gtccgtacga aggtacccag accgctaaac
      241 tgaaagttac caaaggtggt ccgctgccgt tcgcttggga catcctgtcc ccgcagttcc
      301 agtacggttc caaagcttac gttaaacacc cggctgacat cccggactac ctgaaactgt
      361 ccttcccgga aggtttcaaa tgggaacgtg ttatgaactt cgaagacggt ggtgttgtta
      421 ccgttaccca ggactcctcc ctgcaagacg gtgagttcat ctacaaagtt aaactgcgtg
      481 gtaccaactt cccgtccgac ggtccggtta tgcagaaaaa aaccatgggt tgggaagctt
      541 ccaccgaacg tatgtacccg gaagacggtg ctctgaaagg tgaaatcaaa atgcgtctga
      601 aactgaaaga cggtggtcac tacgacgctg aagttaaaac cacctacatg gctaaaaaac
      661 cggttcagct gccgggtgct tacaaaaccg acatcaaact ggacatcacc tcccacaacg
      721 aagactacac catcgttgaa cagtacgaac gtgctgaagg tcgtcactcc accggtgctt
      781 aataacgctg atagtgctag tgtagatcgc tactagagcc aggcatcaaa taaaacgaaa
      841 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta
      901 ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatatac tagtagcggc
      961 cgctgcagtc cggcaaaaaa gggcaaggtg tcaccaccct gccctttttc tttaaaaccg
     1021 aaaagattac ttcgcgttat gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1081 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1141 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1201 aaggccgcgt tgctggcgtt tttccacagg ctccgccccc ctgacgagca tcacaaaaat
     1261 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1321 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1381 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1441 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1501 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1561 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1621 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1681 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1741 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1801 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1861 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1921 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc
     1981 tcgagtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta accaattctg
     2041 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     2101 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     2161 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     2221 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     2281 ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc
     2341 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     2401 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     2461 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     2521 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     2581 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     2641 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     2701 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     2761 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     2821 gcctggagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     2881 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
     2941 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg
     3001 atcagctcga gtgccacttg acgtctaaga aaccattatt atcatgacat taacctataa
     3061 aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga
     3121 tgatttctgg aattcgcggc cgcttctaga gcaatacgca aaccgcctct ccccgcgcgt
     3181 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3241 cgcaacgcaa ttaatgtgag ttagctcact cat
//
