LOCUS       sequence-172967-        3619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A backbone plasmid for creation of additional PUF repeat modules.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3619)
  AUTHORS   Abil Z, Denard CA, Zhao H
  TITLE     Modular assembly of designer PUF proteins for specific 
            post-transcriptional regulation of endogenous RNA.
  JOURNAL   J Biol Eng. 2014 Mar 1;8(1):7.
  PUBMED    24581042
REFERENCE   2  (bases 1 to 3619)
  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: "J Biol 
            Eng."; date: "2014-03-1"; volume: "8(1)"; pages: "7"
COMMENT     Sequence Label: pChlr-pNEB193
FEATURES             Location/Qualifiers
     source          1..3619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1430..2089)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2004..2023
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2090..2192)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(2235..2585)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRFKQSTLDLIKDPARPRVPSSNSLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGICDGR
                     SLRRINKSWCPC"
     primer_bind     2453..2475
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2467..2484
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2468..2484
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2581..2597)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2581..2597)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2594..2616)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2605..2621
                     /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)."
     promoter        complement(join(2629..2635,2636..2653,2654..2659))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2674..2695
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2812..2829)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2983..3571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3063..3082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcgaatt tctgccattc atccgcttat tatcacttat tcaggcgtag caaccaggcg
       61 ctgatgaatc ccctaatgat tttggtaaaa atcattaagt taaggtggat acacatcttg
      121 tcatatgatc aaatggtttc gcgaaaaatc aataatcaga caacaagatg tgcgaactcg
      181 atattttaca cgactctctt taccaattct gccccgaatt acacttaaaa cgactcaaca
      241 gcttaacgtt ggcttgccac gcattacttg actgtaaaac tctcactctt accgaacttg
      301 gccgtaacct gccaaccaaa gcgagaacaa aacataacat caaacgaatc gaccgattgt
      361 taggtaatcg tcacctccac aaagagcgac tcgctgtata ccgttggcat gctagcttta
      421 tctgttcggg caatacgatg cccattgtac ttgttgactg gtctgatatt cgtgagcaaa
      481 aacgacttat ggtattgcga gcttcagtcg cactacacgg tcgttctgtt actctttatg
      541 agaaagcgtt cccgctttca gagcaatgtt caaagaaagc tcatgaccaa tttctagccg
      601 accttgcgag cattctaccg agtaacacca caccgctcat tgtcagtgat gctggcttta
      661 aagtgccatg gtataaatcc gttgagaagc tgggttggta ctggttaagt cgagtaagag
      721 gaaaagtaca atatgcagac ctaggagcgg aaaactggaa acctatcagc aacttacatg
      781 atatgtcatc tagtcactca aagactttag gctataagag gctgactaaa agcaatccaa
      841 tctcatgcca aattctattg tataaatctc gctctaaagg ccgaaaaaat cagcgctcga
      901 cacggactca ttgtcaccac ccgtcaccta aaatctactc agcgtcggca aaggagccat
      961 gggttctagc aactaactta cctgttgaaa ttcgaacacc caaacaactt gttaatatct
     1021 attcgaagcg aatgcagatt gaagaaacct tccgagactt gaaaagtcct gcctacggac
     1081 taggcctacg ccatagccga acgagcagct cagagcgttt tgatatcatg ctgctaatcg
     1141 ccctgatgct tcaactaaca tgttggcttg cgggcgttca tgctcagaaa caaggttggg
     1201 acaagcactt ccaggctaac acagtcagaa atcgaaacgt actctcaaca gttcgcttag
     1261 gcatggaagt tttgcggcat tctggctaca caataacaag ggaagactta ctcgtggctg
     1321 caaccctact agctcaaaat ttattcacac atggttacgc tttggggaaa ttatgagggg
     1381 atctctcaga accaggcgtt taagggcacc aataactgcc tttaaaaaat tacgccccgc
     1441 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     1501 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     1561 aatatttgcc catagtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     1621 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
     1681 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
     1741 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
     1801 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     1861 ttgccatacg gaactccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     1921 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     1981 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     2041 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     2101 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     2161 ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc
     2221 agggcttccc ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg
     2281 tcacaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     2341 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     2401 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     2461 cgacgttgta aaacgacggc cagtgaattc gagctcggta cccgggggcg cgccggatcc
     2521 ttaattaagt ctagagtcga ctgtttaaac ctgcaggcat gcaagcttgg cgtaatcatg
     2581 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2641 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2701 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2761 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2821 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2881 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2941 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3001 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggtgaaacc cgacaggact
     3061 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3121 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3181 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3241 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3301 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3361 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3421 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3481 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3541 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3601 tgacgctcag tggaacgaa
//
