LOCUS       sequence-306382-        3578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the sgRNA targeting to the E. coli can gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3578)
  AUTHORS   Tan SI, Yu PJ, Ng IS
  TITLE     CRISPRi-mediated Programming Essential Gene can as a Direct 
  JOURNAL   Biotechnol Bioeng. 2020 May 27. doi: 10.1002/bit.27443.
  PUBMED    32458463
REFERENCE   2  (bases 1 to 3578)
  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: "Biotechnol 
            Bioeng. 2020 May 27. doi: 10.1002/bit.27443."
COMMENT     Sequence Label: pSUI-SynP-CAS1sg
FEATURES             Location/Qualifiers
     source          1..3578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..44)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    47..68
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    72..88
                     /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)."
     misc_RNA        146..221
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    228..248
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     promoter        295..372
                     /gene="lacI"
                     /label=lacI promoter
     CDS             373..1455
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(392..411)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1653..1670)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1824..2412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1904..1923)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2736..3395)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3310..3329
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     3557..3575
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tcgcggccgc
       61 ttctagagga attgtgagcg gataacaatt ttgacggtgt gagcggataa cagtacagtg
      121 gatcccgttt aaccggtctt gagccgtttt agagctagaa atagcaagtt aaaataaggc
      181 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttac tagtagcggc
      241 cgctgcagtg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
      301 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
      361 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
      421 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
      481 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
      541 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
      601 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
      661 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
      721 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
      781 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
      841 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
      901 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
      961 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     1021 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     1081 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     1141 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     1201 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     1261 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     1321 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     1381 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     1441 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg
     1501 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     1561 catgactatc gtcgccgcac ttatgagatc tattaatgaa tcggccaacg cgcggggaga
     1621 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1681 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1741 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1801 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1861 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1921 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1981 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2041 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2101 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2161 tcgccaccgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2221 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2281 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2341 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2401 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2461 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctc
     2521 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2581 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2641 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2701 tccagtgctg caatgatacc gcgagagtcg acaaattacg ccccgccctg ccactcatcg
     2761 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
     2821 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
     2881 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
     2941 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
     3001 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
     3061 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
     3121 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat
     3181 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
     3241 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
     3301 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     3361 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     3421 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     3481 cctcttacgt gcccgatcaa ctcgagtgcc acctgacgtc taagaaacca ttattatcat
     3541 gacattaacc tataaaaata ggcgtatcac gaggcaga
//
