LOCUS Exported 3404 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Level 0 part. Promoter. ACCESSION . VERSION . KEYWORDS pC.060 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 May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3404 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 78..172 /label=lambda t0 terminator /note="transcription terminator from phage lambda" terminator 328..357 /label=T3Te terminator /note="phage T3 early transcription terminator" promoter 369..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 //