LOCUS       sequence-258462-        3406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 part. Promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3406)
  AUTHORS   Vasudevan R, Gale GAR, Schiavon AA, Puzorjov A, Malin J, Gillespie 
            MD, Vavitsas K, Zulkower V, Wang B, Howe CJ, Lea-Smith DJ, McCormick
            AJ
  TITLE     CyanoGate: A Modular Cloning Suite for Engineering Cyanobacteria 
            Based on the Plant MoClo Syntax.
  JOURNAL   Plant Physiol. 2019 May;180(1):39-55. doi: 10.1104/pp.18.01401. Epub
            2019 Feb 28.
  PUBMED    30819783
REFERENCE   2  (bases 1 to 3406)
  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"; doi: 
            "10.1104/pp.18.01401"; journalName: "Plant Physiol"; date: 
            "2019-05"; volume: "180"; issue: "1"; pages: "39-55"
COMMENT     Sequence Label: pC.061
FEATURES             Location/Qualifiers
     source          1..3406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(78..172)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(328..357)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     promoter        join(369..374,375..392,393..399)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    407..423
                     /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             441..452
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      469..500
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(599..1414)
                     /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     1322..1341
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1406..1425
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(1772..2722)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2770..2992
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caacaggctt acccgtctta
       61 ctgtccctag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg
      121 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag
      181 ctcgtaaact tggtctgaca gctctagctc cggcaaaaaa acgggcaagg tgtcaccacc
      241 ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt tgccacctga cgtctaagaa
      301 aaggaatatt cagcaatttg cccgtgccga agaaaggccc acccgtgaag gtgagccggt
      361 ctcaggagtt gacaattaat catcgcggct cgtataatgt gtggaattgt gagcggataa
      421 caatttcaca catactagag aaagaggaga aatactaaat gtgagaccag tcaaaagcct
      481 ccgaccggag gcttttgact aaaacttccc ttggggttat cattggggct cactcaaagg
      541 cggtaatcag ataaaaaaaa tccttagctt tcgctaagga tgatttctgc tagtattatt
      601 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
      661 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccaaa
      721 gaatggcaag gtcctggtaa cggtctgcga ttccgacccg tccaacatca atacaaccta
      781 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
      841 aatccggtga gaatggcaag agcttgtgca tttctttcca gacttgttca acaggccagc
      901 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatg cgtgattgcg
      961 cctgagcaag acgaaataca cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     1021 gtaaccggcg caggaacacg gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     1081 cttctaatac ctggaaggct gttttcccag gaatcgcggt ggtgagtaac cacgcatcat
     1141 caggagtacg gataaaatgc ttgatggtcg ggagaggcat aaactccgtc agccagttga
     1201 gacggaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     1261 actctggcgc atcgggcttc ccatacaagc gatagattgt cgcacctgat tgcccgacat
     1321 tatcgcgagc ccatttatac ccatataaat cagcgtccat gttggagttt aagcgcggac
     1381 gggagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     1441 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
     1501 gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc
     1561 agctctagta gttacattgt cgatctgttc atggtgaaca gctttgaatg caccaaaaac
     1621 tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt tcatctgtgc atatggacag
     1681 ttttcccttt gatatgtaac ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc
     1741 ttcactgata gatacaagag ccataagaac ctcagatcct tccgtattta gccagtatgt
     1801 tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatact
     1861 tactttgcat gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta
     1921 aagcatcgtg tagtgttttt cttagtccgt tatgtaggta ggaatctgat gtaatggttg
     1981 ttggtatttt gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca
     2041 tttgtctatc tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa
     2101 ccaccaattt catattgctg taagtgttta aatctttact tattggtttc aaaacccatt
     2161 ggttaagcct tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat
     2221 caaggctaat ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc
     2281 actcataaat cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt
     2341 ttatgaattt ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta
     2401 atttttcgct tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca
     2461 aaggattcct gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac
     2521 cataagcatt ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata
     2581 ccgtccgttc tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata
     2641 aaattagctt ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt
     2701 tgaaaacaac taattcagac atacatctca attggtctag gtgattttaa tcactatacc
     2761 aattgagatg ggctagtcaa tgataattac atgtcctttt cctttgagtt gtgggtatct
     2821 gtaaattctg ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt
     2881 ccgctagacc tttgtgtgtt ttttttgttt atattcaagt ggttataatt tatagaataa
     2941 agaaagaata aaaaaagata aaaagaatag atcccagccc tgtgtataac tcactacttt
     3001 agtcagttcc gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca
     3061 gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata
     3121 ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc
     3181 gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg
     3241 attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt
     3301 tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc
     3361 tctgattttc cagtctgacc acttcggatt atcccgtgac aggtca
//
