LOCUS       Exported                2345 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pZE31ms2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2345)
  AUTHORS   Le TT, Harlepp S, Guet CC, Dittmar K, Emonet T, Pan T, Cluzel P
  TITLE     Real-time RNA profiling within a single bacterium.
  JOURNAL   Proc Natl Acad Sci U S A. 2005 Jun 28. 102(26):9160-4.
  PUBMED    15967986
REFERENCE   2  (bases 1 to 2345)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2345
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        6..79
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    6..24
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    31..49
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             89..100
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     111..127
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     111..127
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_RNA        167..185
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        206..224
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(269..285)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(270..286)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             310..345
                     /codon_start=1
                     /product="6x(His-Asn) affinity tag"
                     /label=6xHN
                     /translation="HNHNHNHNHNHN"
     CDS             352..366
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     terminator      443..529
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(693..1281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(773..792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      1369..1463
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1484..2143)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2058..2077
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2144..2246)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2312..2330
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgagtccct atcagtgata gagattgaca tccctatcag tgatagagat actgagcaca
       61 tcagcaggac gcactgaccg aattcattaa agaggagaaa ggtaccccgc tctagaacta
      121 gtggatcccc cgggcagctt gcatgcctgc aggtcgactc tagaaaacat gaggatcacc
      181 catgtctgca ggtcgactct agaaaacatg aggatcaccc atgtctgcag gtcgactcta
      241 gaggatcgga attcgatatc aagcttatcg ataccgtcga cctcgagggg gggcccggta
      301 cccatgggtc ataatcataa tcataatcat aatcataatc acaacggtgg agatgacgat
      361 gacaaggggt cgacaagctt ggatccctgc aggcctcagg gcccgatcga tgcggccgct
      421 taattaatta atctagaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
      481 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta
      541 gacctaggcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      601 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      661 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      721 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      781 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      841 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
      901 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      961 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1021 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1081 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     1141 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1201 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1261 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1321 tggaacgaaa actcacgtta agggattttg gtcatgacta gtgcttggat tctcaccaat
     1381 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
     1441 tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc ccgccctgcc
     1501 actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaga
     1561 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
     1621 tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
     1681 tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag
     1741 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
     1801 gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga
     1861 aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca
     1921 tacggaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa
     1981 acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct
     2041 ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt
     2101 gggatatatc aacggtggta tatccagtga tttttttctc cattttagct tccttagctc
     2161 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     2221 agttggaacc tcttacgtgc cgatcaacgt ctcattttcg ccagatatcg acgtctaaga
     2281 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     2341 tcacc
//
