LOCUS       sequence-174013-        3679 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Encodes module 1 with amino acids 12S-16Q for recognition of nt 8A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3679)
  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 3679)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-1SQ
FEATURES             Location/Qualifiers
     source          1..3679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     71..90
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     324..341
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    458..479
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(494..499,500..517,518..524)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    532..548
                     /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)."
     primer_bind     537..559
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     556..572
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     556..572
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             568..978
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLVSLEDFIEDKLALSWCGTLQFQLNESGPMLVLGKFHY
                     MSVRPSSNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNG
                     EWRLMRYFLLTHLCGICDGRSLRRINKSWCPC"
     primer_bind     complement(729..746)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(729..745)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(738..760)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2462..3121
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2528..2547)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(3261..3679,1..170)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
       61 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcac cacctctgac
      121 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      181 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      241 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
      301 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
      361 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
      421 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
      481 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
      541 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt ggtctctctg
      601 gaggatttca ttgaagacaa gctggcgctc agctggtgtg gcacgctcca gtttcagctg
      661 aatgaatctg gacccatgct ggtcttggga aaattccatt atatgtccgt gagaccgagc
      721 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
      781 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
      841 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
      901 tttctcctta cgcatctgtg cggtatttgt gacggaagat cacttcgcag aataaataaa
      961 tcctggtgtc cctgttgata ccgggaagcc ctgggccaac ttttggcgaa aatgagacgt
     1021 tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgc
     1081 tgagagatcc cctcataatt tccccaaaac gtaaccatgt gtgaatagat tttgagtaag
     1141 cagggttgca gccacgagtg agtcttccct tgttattgtg tagccagaat gccgcaaaac
     1201 ttccatgcct aagcgaactg ttgagagtac gtttcgattt ctgactgtgt tagcctggaa
     1261 gtgcttgtcc caaccttgtt tctgagcatg aacgcccgca agccaacatg ttagttgaag
     1321 catcagggcg attagcagca tgatatcaaa acgctctgag ctgctcgttc ggctatggcg
     1381 taggcctagt ccgtaggcag gacttttcaa gtctcggaag gtttcttcaa tctgcattcg
     1441 cttcgaatag atattaacaa gttgtttggg tgttcgaatt tcaacaggta agttagttgc
     1501 tagaatccat ggctcctttg ccgacgctga gtagatttta ggtgacgggt ggtgacaatg
     1561 agtccgtgtc gagcgctgat tttttcggcc tttagagcga gatttataca atagaatttg
     1621 gcatgagatt ggattgcttt tagtcagcct cttatagcct aaagtctttg agtgactaga
     1681 tgacatatca tgtaagttgc tgataggttt ccagttttcc gctcctaggt ctgcatattg
     1741 tacttttcct cttactcgac ttaaccagta ccaacccagc ttctcaacgg atttatacca
     1801 tggcacttta aagccagcat cactgacaat gagcggtgtg gtgttactcg gtagaatgct
     1861 cgcaaggtcg gctagaaatt ggtcatgagc tttctttgaa cattgctctg aaagcgggaa
     1921 cgctttctca taaagagtaa cagaacgacc gtgtagtgcg actgaagctc gcaataccat
     1981 aagccgtttt tgctcacgga tatcagacca gtcaacaagt acaatgggca tcgtattgcc
     2041 cgaacagata aagctagcat gccaacggta tacagcgagt cgctctttgt ggaggtgacg
     2101 attacctaac aatcggtcga ttcgtttgat gttatgtttt gttctcgctt tggttggcag
     2161 gttacggcca agttcggtaa gagtgagagt tttacagtca agtaaggcgt ggcaagccaa
     2221 cgttaagctg ttgagtcgtt ttaagtgtaa ttcggggcag aattggtaaa gagagtcgtg
     2281 taaaatatcg agttcgcaca ttttgttgtc tgattattga tttttggcga aaccatttga
     2341 tcatatgaca agatgtgtat ctaccttaac ttaatgattt tgataaaaat cattagggga
     2401 ttcatcagta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
     2461 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     2521 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     2581 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     2641 tcacattctt gcccgcctga tgaatgctca tccggagttc cgtatggcaa tgaaagacgg
     2701 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     2761 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     2821 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     2881 gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     2941 ggccaatatg gacaacttct tcgcccccgt tttcactatg ggcaaatatt atacgcaagg
     3001 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     3061 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     3121 attttttaaa ggcagttatt ggtgccctta aacgcctggt tgctacgcct gaataagtga
     3181 taataagcgg atgaatggca gaaattcgaa agttcgttcc actgagcgtc agaccccgta
     3241 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3301 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3361 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3421 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3481 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3541 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3601 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3661 agcgccacgc ttcccgaag
//
