LOCUS       40924_29541        4712 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLZ1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4712)
  AUTHORS   Zhu L, Klenner C, Kuhn A, Dalbey RE.
  TITLE     Both YidC and SecYEG are required for translocation of the 
            periplasmic loops 1 and 2 of the multispanning membrane protein TatC
  JOURNAL   J. Mol. Biol. 424 (5), 354-367 (2012)
  PUBMED    23058713
REFERENCE   2  (bases 1 to 4712)
  AUTHORS   Zhu L, Wasey A, White SH, Dalbey RE.
  TITLE     Charge composition features of model single-span membrane proteins 
            that determine selection of YidC and SecYEG translocase pathways in 
            Escherichia coli
  JOURNAL   J. Biol. Chem. 288 (11), 7704-7716 (2013)
  PUBMED    23355473
REFERENCE   3  (bases 1 to 4712)
  AUTHORS   Zhu L, Kaback HR, Dalbey RE.
  TITLE     YidC protein, a molecular chaperone for LacY protein folding via the
            SecYEG protein machinery
  JOURNAL   J. Biol. Chem. 288 (39), 28180-28194 (2013)
  PUBMED    23928306
REFERENCE   4  (bases 1 to 4712)
  AUTHORS   Soman R, Yuan J, Kuhn A, Dalbey RE.
  TITLE     The polarity and charge of the periplasmic loop determines the YidC 
            and Sec translocase requirement for the M13 procoat lep protein
  JOURNAL   J. Biol. Chem. (2013) In press
  PUBMED    24275657
REFERENCE   5  (bases 1 to 4712)
  AUTHORS   Zhu L, Dalbey R.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-NOV-2013) Department of Chemistry and Biochemistry, 
            The Ohio State University, RM580, W. 12th Ave., Columbus, OH 43210, 
            USA
REFERENCE   6  (bases 1 to 4712)
  TITLE     Direct Submission
REFERENCE   7  (bases 1 to 4712)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Mol. 
            Biol."; date: "2012"; volume: "424"; issue: "5"; pages: "354-367"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "J. Biol. 
            Chem."; date: "2013"; volume: "288"; issue: "11"; pages: "7704-7716"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "J. Biol. 
            Chem."; date: "2013"; volume: "288"; issue: "39"; pages: 
            "28180-28194"
COMMENT     SGRef: number: 4; type: "Journal Article"; journalName: "J. Biol. 
            Chem. (2013) In press"
COMMENT     SGRef: number: 5; type: "Journal Article"; journalName: "Submitted 
            (29-NOV-2013) Department of Chemistry and Biochemistry, The Ohio 
            State University, RM580, W. 12th Ave., Columbus, OH 43210, USA"
COMMENT     SGRef: number: 6; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    39..63
                     /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)."
     promoter        197..227
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    235..251
                     /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     complement(397..413)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      604..651
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      704..790
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      882..909
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      1009..1057
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     terminator      1119..1167
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             1313..1357
                     /codon_start=1
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /translation="KETAAAKFERQHMDS"
     terminator      1409..1456
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1689..2501)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(2502..2593)
                     /label=AmpR promoter
     rep_origin      complement(2609..3358)
                     /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(3516..3537)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3553..4632)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4633..4710)
                     /label=lacI promoter
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
       61 tcccctctag cgcgtattca ggctgaccct gcgcgctgcg cagggcttta ttgattccat
      121 ttttacactg atgaatgttc cgttgcgctg cccggattac aggggaatta attctcactc
      181 attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt ggaattgtga
      241 gcggataaca atttcacaca ggaaacagaa ttaattctta cacttagtta aattgctaac
      301 tttatagatt acaaaactta ggagggtttt taccatgggg gatcccaagc ttcttctaga
      361 ggtaccgcat gcgatatcga gctctcccgg gaattcactg gccgtcgttt tacaacgtcg
      421 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
      481 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
      541 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
      601 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
      661 ggaactatat ccggatatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
      721 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
      781 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
      841 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
      901 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt
      961 atccgctcat gctagaggtc gaaattcacc tcgaaagcaa gctgataaac cgatacaatt
     1021 aaaggctcct tttggagcct ttttttttgg agattttcaa cgtgaaaaaa ttattattcg
     1081 caattccaag ctaattcacc tcgaaagcaa gctgataaac cgatacaatt aaaggctcct
     1141 tttggagcct ttttttttgg agattttcaa cgtgaaaaaa ttattattcg caattccaag
     1201 ctctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     1261 acggtcacag cttgtctgta agcggatgca gacggccgca ctcgagtctg gtaaagaaac
     1321 cgctgctgcg aaatttgaac gccagcacat ggactcgtct actagcgcag cttaattaac
     1381 ctaggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     1441 cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca cactgcttcc
     1501 ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc
     1561 tgaaccgacg acaagctgac gaccgggtct ccgcaagtgg cacttttcgg ggaaatgtgc
     1621 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt
     1681 aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     1741 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     1801 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     1861 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     1921 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     1981 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     2041 gattgcgcct gagcgagacg aaatacgcgg tcgctgttaa aaggacaatt acaaacagga
     2101 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     2161 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     2221 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     2281 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     2341 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     2401 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     2461 cgcggcctag agcaagacgt ttcccgttga atatggctca tactcttcct ttttcaatat
     2521 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2581 aaaaataaac aaataggcat gcagcgctct tccgcttcct cgctcactga ctcgctacgc
     2641 tcggtcgttc gactgcggcg agcggtgtca gctcactcaa aagcggtaat acggttatcc
     2701 acagaatcag gggataaagc cggaaagaac atgtgagcaa aaagcaaagc accggaagaa
     2761 gccaacgccg caggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2821 acgctcaagc cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2881 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2941 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgttggt atctcagttc
     3001 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3061 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3121 actggcagca gccattggta actgatttag aggactttgt cttgaagtta tgcacctgtt
     3181 aaggctaaac tgaaagaaca gattttggtg agtgcggtcc tccaacccac ttaccttggt
     3241 tcaaagagtt ggtagctcag cgaaccttga gaaaaccacc gttggtagcg gtggtttttc
     3301 tttatttatg agatgatgaa tcaatcggtc tatcaagtca acgaacagct attccgttac
     3361 tctagaccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct
     3421 acgtgttccg cttcctttag cagcccttgc gccctgagtg cttgcggcag cgtgaagctg
     3481 gtcggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3541 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3601 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     3661 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     3721 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttgacgg
     3781 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     3841 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     3901 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     3961 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     4021 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     4081 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     4141 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     4201 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     4261 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     4321 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     4381 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     4441 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     4501 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     4561 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     4621 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     4681 gcgaaaggtt ttgcgccatt ctatggtgtc gg
//
