LOCUS       sequence-234053-        3405 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 3405)
  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 3405)
  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.059
FEATURES             Location/Qualifiers
     source          1..3405
                     /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..391,392..398)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    406..422
                     /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             440..451
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      468..499
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(598..1413)
                     /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     1321..1340
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1405..1424
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(1771..2721)
                     /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      2769..2991
                     /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 catccggctc gtataatgtg tggaattgtg agcggataac
      421 aatttcacac atactagaga aagaggagaa atactaaatg tgagaccagt caaaagcctc
      481 cgaccggagg cttttgacta aaacttccct tggggttatc attggggctc actcaaaggc
      541 ggtaatcaga taaaaaaaat ccttagcttt cgctaaggat gatttctgct agtattatta
      601 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
      661 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccaaag
      721 aatggcaagg tcctggtaac ggtctgcgat tccgacccgt ccaacatcaa tacaacctat
      781 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
      841 atccggtgag aatggcaaga gcttgtgcat ttctttccag acttgttcaa caggccagcc
      901 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcatgc gtgattgcgc
      961 ctgagcaaga cgaaatacac gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     1021 taaccggcgc aggaacacgg ccagcgcatc aacaatattt tcacctgaat caggatattc
     1081 ttctaatacc tggaaggctg ttttcccagg aatcgcggtg gtgagtaacc acgcatcatc
     1141 aggagtacgg ataaaatgct tgatggtcgg gagaggcata aactccgtca gccagttgag
     1201 acggaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     1261 ctctggcgca tcgggcttcc catacaagcg atagattgtc gcacctgatt gcccgacatt
     1321 atcgcgagcc catttatacc catataaatc agcgtccatg ttggagttta agcgcggacg
     1381 ggagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     1441 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     1501 attttgagac acaacgtggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca
     1561 gctctagtag ttacattgtc gatctgttca tggtgaacag ctttgaatgc accaaaaact
     1621 cgtaaaagct ctgatgtatc tatctttttt acaccgtttt catctgtgca tatggacagt
     1681 tttccctttg atatgtaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct
     1741 tcactgatag atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt
     1801 ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatactt
     1861 actttgcatg tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa
     1921 agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag gaatctgatg taatggttgt
     1981 tggtattttg tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat
     2041 ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac
     2101 caccaatttc atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg
     2161 gttaagcctt ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc
     2221 aaggctaatc tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca
     2281 ctcataaatc ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt
     2341 tatgaatttt tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa
     2401 tttttcgctt gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa
     2461 aggattcctg atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc
     2521 ataagcattt tccctactga tgttcatcat ctgagcgtat tggttataag tgaacgatac
     2581 cgtccgttct ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa
     2641 aattagcttg gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt
     2701 gaaaacaact aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca
     2761 attgagatgg gctagtcaat gataattaca tgtccttttc ctttgagttg tgggtatctg
     2821 taaattctgc tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc
     2881 cgctagacct ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa
     2941 gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta
     3001 gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag
     3061 accttaaaac cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat
     3121 tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg
     3181 ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga
     3241 ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt
     3301 ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct
     3361 ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtca
//
