LOCUS       sequence-212174-        6027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modified (delta CRP-binding site) 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 6027)
  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 6027)
  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: pCK314
FEATURES             Location/Qualifiers
     source          1..6027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1146..1148,1149..1151,1152..1865)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1191..1212)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1452..1471)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1799..1820
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1971..1990)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1982..2797
                     /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             1982..2797
                     /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(2055..2074)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             2832..3668
                     /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      5040..5585
                     /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     5486..5505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5702..5719
                     /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 gttcatcttt ccctggttgc caatggccca ttttcctgtc agtaacgaga aggtcgcgta
     1081 ttcaggcgct ttttagactg gtcgtaatga atcgggtaag tttataatat acaaaggagg
     1141 tagaaatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1201 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1261 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1321 ccaccctcgt gaccaccttc ggctacggcc tgcaatgctt cgcccgctac cccgaccaca
     1381 tgaagctgca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1441 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1501 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1561 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1621 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1681 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1741 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1801 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1861 agtaaggatc caaagccacg ttgtgtctca aaatctctga tgttacattg cacaagataa
     1921 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     1981 tatgagccat attcaacggg aaacgtcttg ctcgaggccg cgattaaatt ccaacatgga
     2041 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2101 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2161 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2221 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2281 gatccccggg aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2341 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2401 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2461 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2521 agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2581 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2641 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2701 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2761 attgcagttt catttgatgc tcgatgagtt tttctaatat ttcgccgtgt tgacgacatc
     2821 aggaggccag tatgaccgta ttacatagtg tggatttttt tccgtctggt aacgcgtccg
     2881 tggcgataga accccggctc ccgcaggcgg attttcctga acatcatcat gattttcatg
     2941 aaattgtgat tgtcgaacat ggcacgggta ttcatgtgtt taatgggcag ccctatacca
     3001 tcaccggtgg cacggtctgt ttcgtacgcg atcatgatcg gcatctgtat gaacataccg
     3061 ataatctgtg tctgaccaat gtgctgtatc gctcgccgga tcgatttcag tttctcgccg
     3121 ggctgaatca gttgctgcca caagagctgg atgggcagta tccgtctcac tggcgcgtta
     3181 accacagcgt attgcagcag gtgcgacagc tggttgcaca gatggaacag caggaagggg
     3241 aaaatgattt accctcgacc gccagtcgcg agatcttgtt tatgcaatta ctgctcttgc
     3301 tgcgtaaaag cagtttgcag gagaacctgg aaaacagcgc atcacgtctc aacttgcttc
     3361 tggcctggct ggaggaccat tttgccgatg aggtgaattg ggatgccgtg gcggatcaat
     3421 tttctctttc actgcgtacg ctacatcggc agcttaagca gcaaacggga ctgacgcctc
     3481 agcgatacct gaaccgcctg cgactgatga aagcccgaca tctgctacgc cacagcgagg
     3541 ccagcgttac tgacatcgcc tatcgctgtg gattcagcga cagtaaccac ttttcgacgc
     3601 tttttcgccg agagtttaac tggtcaccgc gtgatattcg ccagggacgg gatggctttc
     3661 tgcaataatc ttgggggaaa ttaagaccaa actcgatgac ctccaaaaag atgtaacttc
     3721 tcttaagatc gatatggcaa cggtgaaaac cgagttatct gcggtcagga tggagatagg
     3781 tacagtcaag gatgatgtta aagatgtcaa agggcgggct aatgctcaaa tttgggcgtt
     3841 gattcttgcc gtcatcggag ccataattac caccttggtg cgttttggca ttttccctaa
     3901 tccctaacaa aaaagcgacc aggcttttct ttcaattgcc cgatcgcctt tgatattttc
     3961 ccaaaggata aaagctagtc cattcagaat cgagccttaa agtactccca tattggctag
     4021 ccccagaatt actccagcgc cgaggatgtg gccaaagcta gcggtgccca gcacagcccc
     4081 taaaccaaag ccgccaaaga agttagagga aggcatgggg gtgcccacat tttgttgttt
     4141 gatggtcaat ttaccaaagg cgatcgccaa aatgttgcaa gcaatcatca ccccagcaac
     4201 tttagggctc caggacaggg tggcgggaac ggcggtggcc aacaaaaagc tatgcattga
     4261 gattctccag aataaagacg gtttttaaag ggatagcccc acgctaatgg gggtctttaa
     4321 aaatctcatc ttacggggac gctctgcccc tgggaaacca ccgttgcaat acttaacaaa
     4381 ttttcgtttt tagcttggca aatgtctttg gcaaaattgg ttgatctggc ttaaatcgtc
     4441 agttatttgc cctggaatag tctggggacg ggcaattctg atcagattta cccccaacgc
     4501 ttccgccact ttttgcttaa ccaattctcc cccctgggca ccggaggctt tagttaccac
     4561 cccttgaatt tgccattgtt gccacagggc tttttccaat ggttcggcta cgggggggcg
     4621 caaagcaatg atacggtcgg aagtaaaccc agcggcgatc gcctgggcta gggcttgggg
     4681 atagggcaga atacgggcaa atagaacaaa tctggagccg gtgagcgtgg gtctcgcggt
     4741 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4801 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4861 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4921 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4981 atcccttaac gtgagttttc gttccactga gcgtcagacc ccttaataag atgatcttct
     5041 tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag
     5101 ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga
     5161 ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag
     5221 actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc
     5281 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg
     5341 ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg
     5401 aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag
     5461 gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5521 ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat
     5581 ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc
     5641 caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg
     5701 tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac
     5761 cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca
     5821 acatagtaag ccagtataca ctccgctagc gctgaggtct gcctcgtgaa gaaggtgttg
     5881 ctgactcata ccaggcctga atcgccccat catccagcca gaaagtgagg gagccacggt
     5941 tgatgagagc tttgttgtag gtggaccagt tggtgatttt gaacttttgc tttgccacgg
     6001 aacggtctgc gttgtcggga agatgcg
//
