LOCUS       40924_24762        4885 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHN1_lacUV5, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4885)
  AUTHORS   Niederholtmeyer H, Wolfstadter BT, Savage DF, Silver PA, Way JC.
  TITLE     Engineering cyanobacteria to synthesize and export hydrophilic 
            products
  JOURNAL   Appl. Environ. Microbiol. 76 (11), 3462-3466 (2010)
  PUBMED    20363793
REFERENCE   2  (bases 1 to 4885)
  AUTHORS   Niederholtmeyer H, Wolfstadter BT, Savage DF, Silver PA, Way JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAR-2010) Systems Biology, Harvard Medical School, 200
            Longwood Ave., Boston, MA 02115, USA
REFERENCE   3  (bases 1 to 4885)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4885)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2010"; volume: "76"; issue: "11"; 
            pages: "3462-3466"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-MAR-2010) Systems Biology, Harvard Medical School, 200 Longwood 
            Ave., Boston, MA 02115, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(21..609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      1564..1650
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1742..1769
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     protein_bind    complement(1791..1810)
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        1825..1855
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    1863..1879
                     /label=lac operator
                     /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)."
     regulatory      1888..1902
                     /regulatory_class="ribosome_binding_site"
     misc_feature    1903..1926
                     /label=multiple cloning site
                     /note="multiple cloning site"
     promoter        1934..2036
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             2037..2693
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        2761..2838
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2839..3918
                     /label=lacI
                     /note="lac repressor"
ORIGIN
        1 ctagtcgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
       61 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      121 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      181 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      241 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      301 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      361 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      421 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      481 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      541 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      601 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtc tagttcagcc
      661 agctcgtcgt gatgtcgaaa cccaagccac cctcagaggt gaaggccgct tcgagcacat
      721 cgggaagcgt gtcgacgacg gtgctcggtg cctcgggtaa gagccagata gatgcggtga
      781 cgttcaccgt cgcgatcgtg gcgggaacca cggtcactgc gtccgttaca actcgcacgt
      841 cgtcggccag cacgactgac tcgacggctt cgattagccc gggagaggca aggccatcag
      901 gctcggtaga cagaatgctg attaacacct cgccgggagc agggctgctc acagccgcgt
      961 ccttcacccg tgggtcagcc gtcagcgctt gatagcggta ccaggccgca ccgcctgcgg
     1021 tcgagctgcc cttgatccgc tcgatggtgc gatcgcgaag ctcgccatcc ggctcattcg
     1081 gctgccgcac cacgccgtag aacgcggcaa ggttgtcgag gtcggggccg tttgagtagc
     1141 gaagcagtgt cgcaagaagt gcgtcgttga tccgctgacg caggatcagc tcgcgggctg
     1201 cgcagacctc cagcagcttg atgaccgggt cggattcgag gatggcggtg tagctggcgt
     1261 cacgcgatcg caggtcgtcg atcaagtcct gcaggatcag ttcaaagtcc agcgcttcga
     1321 tgatggtggg cgcgggaatc gtagcaaagt caagaacggt catgagacga ctaagccctc
     1381 cagcgtgata cgcctgccct cggggatgta gtagccgatc aggttcagct cgacttgacc
     1441 agctgcgctg gctgagacga tgcgaacctt ctccagcttc agccgtggct cccagcgatc
     1501 gagcgcttca gctgtggccg ccaccaggtc aacgatgagg gactggttga tcggtctagt
     1561 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     1621 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     1681 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     1741 cagaaggcca tcctgacgga tggccttttt gcgtttctac tctagtttcg aattgtgagc
     1801 gctcacaatt tcgaaacccc aggctttaca ctttatgctt ccggctcgta taatgtgtgg
     1861 aattgtgagc ggataacaat ttcacacaag gaggaaaaac atatgtctag aaagcttgcg
     1921 gccgcgtagg atccgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat
     1981 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg
     2041 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     2101 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     2161 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     2221 ttcttgcccg cctgatgaat gctcatccgg agttccgtat ggcaatgaaa gacggtgagc
     2281 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     2341 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     2401 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     2461 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     2521 atatggacaa cttcttcgcc cccgttttca ctatgggcaa atattatacg caaggcgaca
     2581 aggtgctgat gccgctggcg attcaggttc atcatgccgt ctgtgatggc ttccatgtcg
     2641 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt
     2701 tttaaggcag ttattggtgc ccttgaattc ctactagtcg aagcggcatg catttacgtt
     2761 gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
     2821 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
     2881 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
     2941 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
     3001 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
     3061 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
     3121 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
     3181 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
     3241 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
     3301 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
     3361 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
     3421 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
     3481 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
     3541 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
     3601 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
     3661 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
     3721 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     3781 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     3841 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3901 cgactggaaa gcgggcagtg atcccacagc cgccagttcc gctggcggca ttttaacttt
     3961 ctttaatgaa tctagtgaca agccggggca gacgtgagcc gtagtcccgt cgccagacgc
     4021 gggtgcccac gggcgtcgtc aggatgtccg taattgactg ccggaggtgg tcaatgccct
     4081 tcagctcctt gccactgtca cggctcatgc ctcgggtcat tagtcgcccg ctccggtatc
     4141 ttcactggct tcgatgattg ccgccccgca gctgcagagg tcaccgatcc gagcagtcgg
     4201 cctctggttg gtaaagaccg tgcgactgcc ggtgatgatc gtgttcaagc catgcagggg
     4261 gcaggcgtga aggtcatcct ttcgggccac ggggcggctg ttgacgaagg tgtcgtcgct
     4321 cccggtgatg atgatcccgc cgtgatcggt cacgtcgttt agtcgagcga tgcctggcgt
     4381 cgtagtcacg ggtttaggtc aatacgactt gcggtcactg taacgttgcc ctcggcggtc
     4441 acgttaacgt cgccttgggc ttcgacttgc gcctcctgca caaggatcac aatccgtcct
     4501 tgggctgcgg tgaggtcgat cttgtactca tgcgcttcgc ggtcgtactg gatgattgag
     4561 tcatcctcga actgcgtctt ttggatcgtt tctttgtcct cgatctgggg gtagtcagtc
     4621 gagaacgcgc cgggcatcgc gaagccctga ctgatctcgc cggagggggc catcacgacg
     4681 acggcctcac cgacctcggg cgcccaccag aaccgatcct tgcccgctcg ctgcgtgagc
     4741 cacggaatcc agtcagtgag gagcagcgcc tcgccgctct cctcgtcttc ctcgatcgcg
     4801 acacggatca gccccttggg atagtcagcc tcggctaccc tgcctacgcg gagcaagttg
     4861 ccgtgacgcc gactgtctcg agtat
//
