LOCUS       V004867                 9272 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004867
VERSION     V004867
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9272)
  AUTHORS   Zhu L, Dalbey RE.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAR-2014) Department of Chemistry and Biochemistry,
            The Ohio State University, RM580, W.12th Ave, Columbus, OH 43210,
            USA
REFERENCE   2  (bases 1 to 9272)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9272)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (31-MAR-2014) Department of Chemistry and Biochemistry, The Ohio
            State University, RM580, W.12th Ave, Columbus, OH 43210, USA"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             28..72
                     /label="S-Tag"
                     /note="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
     terminator      124..171
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(206..2854)
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
                     /label="T7 RNA polymerase"
     CDS             complement(2953..3018)
                     /label="lambda N peptide"
                     /note="N-terminal RNA-binding domain from the lambda
                     bacteriophage antiterminator protein N (Baron-Benhamou et
                     al., 2004)"
     regulatory      complement(3226..3236)
                     /note="pL; induces expression of T7 RNA polymerase"
                     /regulatory_class="promoter"
     regulatory      complement(3247..3252)
                     /label="lambda left promoter"
                     /note="lambda left promoter"
                     /regulatory_class="minus_10_signal"
     regulatory      complement(3270..3275)
                     /label="lambda left promoter"
                     /note="lambda left promoter"
                     /regulatory_class="minus_35_signal"
     promoter        3415..3445
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    3453..3469
                     /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)."
     CDS             3490..4200
                     /label="lambda repressor (ts)"
                     /note="temperature-sensitive variant of the phage lambda
                     repressor"
     CDS             complement(4656..5468)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(5469..5560)
                     /label="AmpR promoter"
     rep_origin      complement(5576..6325)
                     /direction=LEFT
                     /label="RSF ori"
                     /note="Plasmids containing the RSF 1030 origin of
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     protein_bind    complement(6487..6508)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6524..7603)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(7604..7680)
                     /label="lacI promoter"
     promoter        7698..7716
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7781..7802
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7817..7847
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    7855..7871
                     /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     7879..7895
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     complement(7911..7927)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      8007..8013
                     /regulatory_class="ribosome_binding_site"
     CDS             8021..9271
                     /gene="lacY"
                     /label="Lactose permease"
                     /note="Lactose permease from Escherichia coli (strain K12).
                     Accession#: P02920"
ORIGIN
        1 aagcttgcgg ccgcactcga gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag
       61 cacatggact cgtctactag cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa
      121 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
      181 tccgggtcgt attgatttgg cgttacgcga acgcgaagtc cgactctaag atgtcacgga
      241 ggttcaagtt acctttagcc ggaagtgctg gcattttgtc caattgagac tcgtgcaact
      301 ggtcagcgaa ctggtcgtag aaatcagcca gtacatcaca agactcatat gtgtcaacca
      361 tagtttcgcg cactgctttg aacaggttcg cagcgtcagc cggaatggta ccgaaggagt
      421 cgtgaatcag tgcaaaagat tcgattccgt acttctcgtg tgcccacact acagtcttac
      481 gaaggtggct accgtcttgg ctgtgtacaa agttaggagc gataccagac tcctgtttgt
      541 gtgcatcaat ctcgctatct ttgttggtgt taatggtagg ctgtaagcgg aactgaccga
      601 ggaacatcag gttcaagcgc gtctgaatag gcttcttgta ttcctgccac acagggaaac
      661 catcaggagt tacccaatgc acagcgcaac gcttgcgaag aatctctcca gtcttcttat
      721 ctttgacctc agcagccagc agcttagcag cagacttaag ccagttcatt gcttcaaccg
      781 cagctaccac cgtcacgctc acagattccc aaatcagctt agccatgtat ccagcagcct
      841 gattcggctg agtgaacatc agacccttgc cggaatcaat agctggctga atggtatctt
      901 ccagcacttg ttgacggaag ccgaactctt tggacccgta agccagcgtc atgactgaac
      961 gcttagtcac actgcgagta acaccgtaag ccagccattg accagccagt gccttagtgc
     1021 ccagcttgac tttctcagag atttcaccag tgttctcatc ggtcacggta actacttcgt
     1081 tatcggtccc attgattgcg tctgcttgta gaatctcgtt gactttctta gcaacaatcc
     1141 cgtagatgtc ctgaacggtt tcactaggaa gcaagttaac cgcgcgacca cctacctcat
     1201 ctcggagcat cgcggagaag tgctggatgc cagagcaaga cccgtcaaac gccagcggaa
     1261 gggagcagtt atagctcagg ccgtggtgct gtaccccagc gtactcaaag cagaacgcaa
     1321 ggaagcagaa cggagaatct tgctcagccc accaagtgtt ctccagtgga gacttagcgc
     1381 aagccatgat gttctcgtgg ttttcctcaa tgaacttgat gcgctcaggg aacggaacct
     1441 tatcgacacc cgcacagttt gcaccgtgga ttttcagcca gtagtaacct tccttaccga
     1501 ttggtttacc tttcgccagc gtaagcagtc ctttggtcat atcgttacct tgcgggttga
     1561 acattgacac agcgtaaaca cgaccgcgcc agtccatgtt gtaagggaac cagatggcct
     1621 tatggttagc aaacttattg gcttgctcaa gcatgaactc aaggctgata cggcgagact
     1681 tgcgagcctt gtccttgcgg tacacagcag cggcagcacg tttccacgcg gtgagagcct
     1741 caggattcat gtcgatgtct tccggtttca tcgggagttc ttcacgctca atcgcaggga
     1801 tgtcctcgac cggacaatgc ttccacttgg tgattacgtt ggcgaccgct aggactttct
     1861 tgttgatttt ccatgcggtg ttttgcgcaa tgttaatcgc tttgtacacc tcaggcatgt
     1921 aaacgtcttc gtagcgcatc agtgctttct tactgtgagt acgcaccagc gccagaggac
     1981 gacgaccgtt agcccaatag ccaccaccag taatgccagt ccacggctta ggaggaacta
     2041 cgcaaggttg gaacatcgga gagatgccag ccagcgcacc tgcacgggtt gcgatagcct
     2101 cagcgtattc aggtgcgagt tcgatagtct cagagtcttg acctactacg ccagcatttt
     2161 ggcggtgtaa gctaaccatt ccggttgact caatgagcat ctcgatgcag cgtactccta
     2221 catgaataga gtcttcctta tgccacgaag accacgcctc gccaccgagt agacccttag
     2281 agagcatgtc agcctcgaca acttgcataa atgctttctt gtagacgtgc cctacgcgct
     2341 tgttgagttg ttcctcaacg tttttcttga agtgcttagc ttcaaggtca cggatacgac
     2401 cgaagcgagc ctcgtcctca atggcccgac cgattgcgct tgctacagcc tgaacggttg
     2461 tattgtcagc actggttagg caagccagag tggtcttaat ggtgatgtac gctacggctt
     2521 ccggcttgat ttcttgcagg aactggaagg ctgtcgggcg cttgccgcgc ttagctttca
     2581 cttcctcaaa ccagtcgttg atgcgtgcaa tcatcttagg gagtagggta gtgatgagag
     2641 gcttggcggc agcgttatcc gcaacctcac cagctttaag ttgacgctca aacatcttgc
     2701 ggaagcgtgc ttcacccatc tcgtaagact catgctcaag ggccaactgt tcgcgagcta
     2761 aacgctcacc gtaatggtca gccagagtgt tgaacgggat agcagccagt tcgatgtcag
     2821 agaagtcgtt cttagcgatg ttaatcgtgt tcatttagtg cctcttccag ttagtaaatc
     2881 gtacctattc aggccacccg cagatgacct gtaagataaa actggttttg cgcttacccc
     2941 aaccaacagg ggatttgctg ctttccattg agcctgtttc tctgcgcgac gttcgcggcg
     3001 gcgtgtttgt gcatccatct ggattctcct gtcagttagc tttggtggtg tgtggcagtt
     3061 gtagtcctga acgaaaaccc cccgcgattg gcacattggc agctaatccg gaatcgcact
     3121 tacggccaat gcttcgtttc gtatcacaca ccccaaagcc ttctgctttg aatgctgccc
     3181 ttcttcaggg cttaattttt aagagcgtca ccttcatggt ggtcagtgcg tcctgctgat
     3241 gtgctcagta tcaccgccag tggtatttat gtcaacaccg ccagagataa tttatcaccg
     3301 cagatggtta tctgtatgtt ttttatatga atttattttt tgcagggggg cattgtttgg
     3361 taggtgagag atccccgggc gagctcgaat tctcactcat taggcacccc aggctttaca
     3421 ctttatgctt ccggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg
     3481 aaacagcgta tgagcacaaa aaagaaacca ttaacacaag agcagcttga ggacgcacgt
     3541 cgccttaaag caatttatga aaaaaagaaa aatgaacttg gcttatccca ggaatctgtc
     3601 gcagacaaga tggggatggg gcagtcaggc gttggtgctt tatttaatgg catcaatgca
     3661 ttaaatgctt ataacgccgc attgcttaca aaaattctca aagttagcgt tgaagaattt
     3721 agcccttcaa tcgccagaga aatctacgag atgtatgaag cggttagtat gcagccgtca
     3781 cttagaagtg agtatgagta ccctgttttt tctcatgttc aggcagggat gttctcacct
     3841 gagcttagaa cctttaccaa aggtgatgcg gagagatggg taagcacaac caaaaaagcc
     3901 agtgattctg cattctggct tgaggttgaa ggtaattcca tgaccgcacc aacaggctcc
     3961 aagccaagct ttcctgacgg aatgttaatt ctcgttgacc ctgagcaggc tgttgagcca
     4021 ggtgatttct gcatagccag acttgggggt gatgagttta ccttcaagaa actgatcagg
     4081 gatagcggtc aggtgttttt acaaccacta aacccacagc acccaatgat cccatgcaat
     4141 gagagttgtt ccgttgtggg gaaagttatc gctagtcagt ggcctgaaga gacgtttggc
     4201 tgatcggcaa ggtgttctgg tcggcgcata gctgataaca attgagcaag aatcttcatc
     4261 gaattagggg aattttcact cccctcagaa cataacatag taaatggatt gaattatgaa
     4321 gaatggtttt tatgcgactt accgcagcaa aaataaaggg aaagataagc gctcaataaa
     4381 cctgtctgtt ttccttaatt ctctgctggc tgataatcat cacctgcagc ctcaggcatt
     4441 tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac
     4501 ataagcggct atttaacgac cctgccctga accgacgaca agctgacgac cgggtctccg
     4561 caagtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4621 attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga
     4681 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     4741 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     4801 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     4861 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
     4921 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     4981 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcggtcg
     5041 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     5101 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     5161 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     5221 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     5281 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     5341 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     5401 aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata
     5461 tggctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5521 agcggataca tatttgaatg tatttagaaa aataaacaaa taggcatgca gcgctcttcc
     5581 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggtgtcagct
     5641 cactcaaaag cggtaatacg gttatccaca gaatcagggg ataaagccgg aaagaacatg
     5701 tgagcaaaaa gcaaagcacc ggaagaagcc aacgccgcag gcgtttttcc ataggctccg
     5761 cccccctgac gagcatcaca aaaatcgacg ctcaagccag aggtggcgaa acccgacagg
     5821 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5881 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5941 tagctcacgc tgttggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6001 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6061 caacccggta agacacgact tatcgccact ggcagcagcc attggtaact gatttagagg
     6121 actttgtctt gaagttatgc acctgttaag gctaaactga aagaacagat tttggtgagt
     6181 gcggtcctcc aacccactta ccttggttca aagagttggt agctcagcga accttgagaa
     6241 aaccaccgtt ggtagcggtg gtttttcttt atttatgaga tgatgaatca atcggtctat
     6301 caagtcaacg aacagctatt ccgttactct agatttcagt gcaatttatc tcttcaaatg
     6361 tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag tcatgccccg
     6421 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     6481 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6541 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6601 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     6661 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     6721 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     6781 cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc
     6841 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     6901 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     6961 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     7021 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     7081 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     7141 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     7201 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     7261 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     7321 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     7381 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     7441 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     7501 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     7561 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     7621 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgg
     7681 tcgacgctta tcgaaattaa tacgactcac tatagggaga ccggaattcc tggcacgaca
     7741 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     7801 attagggacc cccggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     7861 gcggataaca atttcacaca ggaaacagct atgaccatga ttacggattc actggccgtc
     7921 gttttacaac gtcgtgactg gtaaaacccg ggcgttaccc aacttaatcg ccttgcagca
     7981 catccccctt tcgccagcag gcgtaataag gaaaggatcc atgtactatt tgaaaaacac
     8041 aaacttttgg atgttcggtt tattcttttt cttttacttt tttatcatgg gagcctactt
     8101 cccgtttttc ccgatttggc tacatgatat caaccatatc agcaaaagtg atacgggtat
     8161 tatttttgcc gctatttctc tgttctcgct attattccaa ccgctgtttg gtctgctttc
     8221 tgacaaactc ggtctacgca aatacctgct gtggattatt accggcatgt tagtgatgtt
     8281 tgcgccgttc tttattttta tcttcgggcc actgctgcag tacaacattt tagtaggatc
     8341 gattgttggt ggtatttatc taggctttag ttttaacgcc ggtgcgccag cagtagaggc
     8401 atttattgag aaagtcagcc ggcgcagtaa tttcgaattt ggtcgcgcgc ggatgtttgg
     8461 cagtgttggc tgggcgctgg ttgcctcgat tgtcgggatc atgttcacca ttaataatca
     8521 gtttgttttc tggctgggct ctggcagtgc actcatcctc gccgttttac tctttttcgc
     8581 caaaacggac gcgccctcga gtgccacggt tgccaatgcg gtaggtgcca accattcggc
     8641 atttagcctt aagctggcac tggaactgtt cagacagcca aaactgtggt ttttgtcact
     8701 gtatgttatt ggcgtttcct ccacctacga tgtttttgac caacagtttg ctaatttctt
     8761 tacttcgttc tttgctaccg gtgaacaggg tacccgcgta tttggctacg taacgacaat
     8821 gggcgaatta cttaacgcct cgattatgtt ctttgcgcca ctgatcatta atcgcatcgg
     8881 tgggaagaat gccctgctgc tggctggcac tattatgtct gtacgtatta ttggctcatc
     8941 gttcgccacc tcagcgctgg aagtggttat tctgaaaacg ctgcatatgt ttgaagtacc
     9001 gttcctgctg gtgggctcct ttaaatatat tactagtcag tttgaagtgc gtttttcagc
     9061 gacgatttat ctggtcagct tcagcttctt taagcaactg gcgatgattt ttatgtctgt
     9121 actggcgggc aatatgtatg aaagcatagg tttccaaggc gcttatctgg tgctgggtct
     9181 ggtggcgctg ggcttcacct taatttccgt gttcacgctt agcggcccgg gcccgctttc
     9241 cctgctgcgt cgtcaggtga atgaagtcgc tt
//
