LOCUS       sequence-258458-        3404 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 3404)
  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 3404)
  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.060
FEATURES             Location/Qualifiers
     source          1..3404
                     /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..390,391..397)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    405..421
                     /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             439..450
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      467..498
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(597..1412)
                     /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     1320..1339
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1404..1423
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(1770..2720)
                     /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      2768..2990
                     /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 catcggctcg tataatgtgt ggaattgtga gcggataaca
      421 atttcacaca tactagagaa agaggagaaa tactaaatgt gagaccagtc aaaagcctcc
      481 gaccggaggc ttttgactaa aacttccctt ggggttatca ttggggctca ctcaaaggcg
      541 gtaatcagat aaaaaaaatc cttagctttc gctaaggatg atttctgcta gtattattag
      601 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
      661 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccaaaga
      721 atggcaaggt cctggtaacg gtctgcgatt ccgacccgtc caacatcaat acaacctatt
      781 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
      841 tccggtgaga atggcaagag cttgtgcatt tctttccaga cttgttcaac aggccagcca
      901 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcatgcg tgattgcgcc
      961 tgagcaagac gaaatacacg atcgctgtta aaaggacaat tacaaacagg aatcgaatgt
     1021 aaccggcgca ggaacacggc cagcgcatca acaatatttt cacctgaatc aggatattct
     1081 tctaatacct ggaaggctgt tttcccagga atcgcggtgg tgagtaacca cgcatcatca
     1141 ggagtacgga taaaatgctt gatggtcggg agaggcataa actccgtcag ccagttgaga
     1201 cggaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     1261 tctggcgcat cgggcttccc atacaagcga tagattgtcg cacctgattg cccgacatta
     1321 tcgcgagccc atttataccc atataaatca gcgtccatgt tggagtttaa gcgcggacgg
     1381 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     1441 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
     1501 ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag
     1561 ctctagtagt tacattgtcg atctgttcat ggtgaacagc tttgaatgca ccaaaaactc
     1621 gtaaaagctc tgatgtatct atctttttta caccgttttc atctgtgcat atggacagtt
     1681 ttccctttga tatgtaacgg tgaacagttg ttctactttt gtttgttagt cttgatgctt
     1741 cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc cagtatgttc
     1801 tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag atcatactta
     1861 ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt tgcagttaaa
     1921 gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg aatctgatgt aatggttgtt
     1981 ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac gagatccatt
     2041 tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg cttatcaacc
     2101 accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa aacccattgg
     2161 ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt aaattcatca
     2221 aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt taataaccac
     2281 tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag attatatttt
     2341 atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac taattctaat
     2401 ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc tttaaccaaa
     2461 ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc taatacacca
     2521 taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt gaacgatacc
     2581 gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac acagcataaa
     2641 attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc atttgctttg
     2701 aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc actataccaa
     2761 ttgagatggg ctagtcaatg ataattacat gtccttttcc tttgagttgt gggtatctgt
     2821 aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc tgtaaattcc
     2881 gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta tagaataaag
     2941 aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc actactttag
     3001 tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct acaaaacaga
     3061 ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc gctgaatatt
     3121 ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc
     3181 tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc ttttatggat
     3241 tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct cagggcgttt
     3301 tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc
     3361 tgattttcca gtctgaccac ttcggattat cccgtgacag gtca
//
