LOCUS       sequence-212164-        6059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  rhaBAD, a rhamnose-inducible promoter, YFP, an E. coli-Synechocystis
            shuttle vector, chromosomal-integration sites. Allows precise.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6059)
  AUTHORS   Kelly CL, Taylor GM, Hitchcock A, Torres-Mendez A, Heap JT
  TITLE     A Rhamnose-Inducible System for Precise and Temporal Control of Gene
            Expression in Cyanobacteria.
  JOURNAL   ACS Synth Biol. 2018 Apr 2. doi: 10.1021/acssynbio.7b00435.
  PUBMED    29544054
REFERENCE   2  (bases 1 to 6059)
  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: "ACS Synth 
            Biol. 2018 Apr 2. doi: 10.1021/acssynbio.7b00435."
COMMENT     Sequence Label: pCK306
FEATURES             Location/Qualifiers
     source          1..6059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1178..1180,1181..1183,1184..1897)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1223..1244)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1484..1503)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1831..1852
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2003..2022)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2014..2829
                     /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             2014..2829
                     /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     complement(2087..2106)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             2864..3700
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     rep_origin      5072..5617
                     /direction=RIGHT
                     /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     5518..5537
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5734..5751
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtgggtcaat ccctcggttt ggcgatcgcc ttggtgtttg tcattgctgg tattgccttt
       61 aaaatttccg ctgtaccctt ccaccaatgg acccccgacg tttatgaagg ttctcccacc
      121 ccggtggtgg cttttctttc ggtgggctct aaggcggccg gttttgccgt agctatccga
      181 ttactagtga ctgcctttgg tggtattacc gatgagtggc acgttatttt caccgctctg
      241 gccgtgttga gcatggtgct gggcaacgtg gtggctttag cccaaaccag catgaaacgg
      301 atgttggcct actcttccat cggtcaagca ggctttgtga tgattggcct agtggccggc
      361 agtgaagatg gttacgccag catggttttc tacatgctca tctatctgtt tatgaacctg
      421 ggggcgttta gttgcattat tctcttcacc ctccgcactg gcagtgacca aattagtgat
      481 tacgctggtc tgtaccacaa agaccccttg ttaaccttgg gcttgagcat ttgtttatta
      541 tccttggggg gcattcctcc tctggcgggc tttttcggca aaatttacat cttctgggcc
      601 ggttggcaat cgggattgta tggcctagtc ctacttggtc tggttaccag tgtagtttcc
      661 atctactact acatccgggt ggtgaaaatg atggtggtga aggagcccca ggaaatgtcc
      721 gaagtaatca aaaattaccc ggccatcaaa tggaatttac ccggcatgcg tcccctacag
      781 gtgggcattg tcgctacttt ggttgctacc tcgctggcag gtattctggc taatcccctc
      841 tttaacctcg ccaccgattc cgtggtcagc accaagatgt tgcagacagc cctccagcaa
      901 acaggagaaa ctccggcgat cgccatttcc catgatttac cctaggggta tcaggaaata
      961 ttgctttgca ggcaaaagcc aatgagtgta actatagaaa ccgatttaaa ggagactgca
     1021 gccacaattc agcaaattgt gaacatcatc acgttcatct ttccctggtt gccaatggcc
     1081 cattttcctg tcagtaacga gaaggtcgcg tattcaggcg ctttttagac tggtcgtaat
     1141 gaatcgggta agtttataat atacaaagga ggtagaaatg gtgagcaagg gcgaggagct
     1201 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1261 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1321 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg
     1381 cctgcaatgc ttcgcccgct accccgacca catgaagctg cacgacttct tcaagtccgc
     1441 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1501 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1561 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1621 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1681 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1741 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
     1801 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1861 cgggatcact ctcggcatgg acgagctgta caagtaagga tccaaagcca cgttgtgtct
     1921 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     1981 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct
     2041 tgctcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     2101 cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg
     2161 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2221 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     2281 actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta
     2341 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     2401 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     2461 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     2521 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataagctttt gccattctca
     2581 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     2641 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     2701 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2761 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2821 tttttctaat atttcgccgt gttgacgaca tcaggaggcc agtatgaccg tattacatag
     2881 tgtggatttt tttccgtctg gtaacgcgtc cgtggcgata gaaccccggc tcccgcaggc
     2941 ggattttcct gaacatcatc atgattttca tgaaattgtg attgtcgaac atggcacggg
     3001 tattcatgtg tttaatgggc agccctatac catcaccggt ggcacggtct gtttcgtacg
     3061 cgatcatgat cggcatctgt atgaacatac cgataatctg tgtctgacca atgtgctgta
     3121 tcgctcgccg gatcgatttc agtttctcgc cgggctgaat cagttgctgc cacaagagct
     3181 ggatgggcag tatccgtctc actggcgcgt taaccacagc gtattgcagc aggtgcgaca
     3241 gctggttgca cagatggaac agcaggaagg ggaaaatgat ttaccctcga ccgccagtcg
     3301 cgagatcttg tttatgcaat tactgctctt gctgcgtaaa agcagtttgc aggagaacct
     3361 ggaaaacagc gcatcacgtc tcaacttgct tctggcctgg ctggaggacc attttgccga
     3421 tgaggtgaat tgggatgccg tggcggatca attttctctt tcactgcgta cgctacatcg
     3481 gcagcttaag cagcaaacgg gactgacgcc tcagcgatac ctgaaccgcc tgcgactgat
     3541 gaaagcccga catctgctac gccacagcga ggccagcgtt actgacatcg cctatcgctg
     3601 tggattcagc gacagtaacc acttttcgac gctttttcgc cgagagttta actggtcacc
     3661 gcgtgatatt cgccagggac gggatggctt tctgcaataa tcttggggga aattaagacc
     3721 aaactcgatg acctccaaaa agatgtaact tctcttaaga tcgatatggc aacggtgaaa
     3781 accgagttat ctgcggtcag gatggagata ggtacagtca aggatgatgt taaagatgtc
     3841 aaagggcggg ctaatgctca aatttgggcg ttgattcttg ccgtcatcgg agccataatt
     3901 accaccttgg tgcgttttgg cattttccct aatccctaac aaaaaagcga ccaggctttt
     3961 ctttcaattg cccgatcgcc tttgatattt tcccaaagga taaaagctag tccattcaga
     4021 atcgagcctt aaagtactcc catattggct agccccagaa ttactccagc gccgaggatg
     4081 tggccaaagc tagcggtgcc cagcacagcc cctaaaccaa agccgccaaa gaagttagag
     4141 gaaggcatgg gggtgcccac attttgttgt ttgatggtca atttaccaaa ggcgatcgcc
     4201 aaaatgttgc aagcaatcat caccccagca actttagggc tccaggacag ggtggcggga
     4261 acggcggtgg ccaacaaaaa gctatgcatt gagattctcc agaataaaga cggtttttaa
     4321 agggatagcc ccacgctaat gggggtcttt aaaaatctca tcttacgggg acgctctgcc
     4381 cctgggaaac caccgttgca atacttaaca aattttcgtt tttagcttgg caaatgtctt
     4441 tggcaaaatt ggttgatctg gcttaaatcg tcagttattt gccctggaat agtctgggga
     4501 cgggcaattc tgatcagatt tacccccaac gcttccgcca ctttttgctt aaccaattct
     4561 cccccctggg caccggaggc tttagttacc accccttgaa tttgccattg ttgccacagg
     4621 gctttttcca atggttcggc tacggggggg cgcaaagcaa tgatacggtc ggaagtaaac
     4681 ccagcggcga tcgcctgggc tagggcttgg ggatagggca gaatacgggc aaatagaaca
     4741 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4801 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4861 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4921 tttactaata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4981 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5041 gagcgtcaga ccccttaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct
     5101 cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta
     5161 ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt
     5221 cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg
     5281 gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg
     5341 gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga
     5401 actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg
     5461 aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg
     5521 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat
     5581 ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc
     5641 acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg ttcgtaagcc
     5701 atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa
     5761 tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct cctgccacat
     5821 gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata cactccgcta
     5881 gcgctgaggt ctgcctcgtg aagaaggtgt tgctgactca taccaggcct gaatcgcccc
     5941 atcatccagc cagaaagtga gggagccacg gttgatgaga gctttgttgt aggtggacca
     6001 gttggtgatt ttgaactttt gctttgccac ggaacggtct gcgttgtcgg gaagatgcg
//
