LOCUS       sequence-252123-        3741 bp    DNA     circular SYN 15-DEC-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 3741)
  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 3741)
  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: "2018-05-10"; volume: "12:8. doi"; pages: " 
            10.1186/s13036-018-0100-0. eCollection 2018"
COMMENT     Sequence Label: pSB3K3 containing mRFP1
FEATURES             Location/Qualifiers
     source          1..3741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(17..60)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    63..84
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    185..206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(221..226,227..244,245..251)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    259..275
                     /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             293..304
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             311..988
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1033..1104
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1120..1147
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1154..1174
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(1314..2129)
                     /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(1314..2129)
                     /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     2037..2056
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2121..2140
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(2724..3268)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2803..2822)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(3736..3741,1..13)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa ggatgatttc
       61 tggaattcgc ggccgcttct agagcaatac gcaaaccgcc tctccccgcg cgttggccga
      121 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
      181 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
      241 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacatactag agaaagagga
      301 gaaatactag atggcttcct ccgaagacgt tatcaaagag ttcatgcgtt tcaaagttcg
      361 tatggaaggt tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc
      421 gtacgaaggt acccagaccg ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc
      481 ttgggacatc ctgtccccgc agttccagta cggttccaaa gcttacgtta aacacccggc
      541 tgacatcccg gactacctga aactgtcctt cccggaaggt ttcaaatggg aacgtgttat
      601 gaacttcgaa gacggtggtg ttgttaccgt tacccaggac tcctccctgc aagacggtga
      661 gttcatctac aaagttaaac tgcgtggtac caacttcccg tccgacggtc cggttatgca
      721 gaaaaaaacc atgggttggg aagcttccac cgaacgtatg tacccggaag acggtgctct
      781 gaaaggtgaa atcaaaatgc gtctgaaact gaaagacggt ggtcactacg acgctgaagt
      841 taaaaccacc tacatggcta aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat
      901 caaactggac atcacctccc acaacgaaga ctacaccatc gttgaacagt acgaacgtgc
      961 tgaaggtcgt cactccaccg gtgcttaata acgctgatag tgctagtgta gatcgctact
     1021 agagccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     1081 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
     1141 ttctgcgttt atatactagt agcggccgct gcaggcttcc tcgctcactg actcgctgcg
     1201 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tctcgagtcc
     1261 cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa
     1321 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc gataccatat
     1381 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     1441 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     1501 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     1561 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
     1621 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     1681 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     1741 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     1801 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     1861 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     1921 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     1981 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     2041 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctggag
     2101 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     2161 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     2221 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc
     2281 acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac
     2341 tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat
     2401 ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag acctcagcgc
     2461 tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc
     2521 atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata
     2581 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat
     2641 ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta acagggaagt
     2701 gagagggccg cggcaaagcc gtttttccat aggctccgcc ccctgacaag catcacgaaa
     2761 tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2821 cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc
     2881 cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt
     2941 cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc
     3001 ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc
     3061 actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga
     3121 aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt
     3181 agctcagaga accttcaaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga
     3241 gattacgcgc agaccaaaac gatctcaaga agatcatctt attaaggggt ctgacgctca
     3301 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3361 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3421 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3481 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3541 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3601 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3661 cgcctccatc cagtctattc catggtgcca cctgacgtct aagaaaccat tattatcatg
     3721 acattaacct ataaaaatag g
//
