LOCUS       40924_48393        5832 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pZH3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5832)
  AUTHORS   Hu JC.
  TITLE     Repressor fusions as a tool to study protein-protein interactions
  JOURNAL   Structure 3 (5), 431-433 (1995)
  PUBMED    7663940
REFERENCE   2  (bases 1 to 5832)
  AUTHORS   Zhu H, Celinski SA, Scholtz JM, Hu JC.
  TITLE     The contribution of buried polar groups to the conformational 
            stability of the GCN4 coiled coil
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5832)
  AUTHORS   Zhu H, Celinski SA, Scholtz JM, Hu JC.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   4  (bases 1 to 5832)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5832)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Structure";
            date: "1995"; volume: "3"; issue: "5"; pages: "431-433"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (13-JUL-1999) Biochemistry "
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        12..40
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     terminator      complement(408..455)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(525..590)
                     /codon_start=1
                     /label=GCN4_v1
                     /note="GCN4 peptide that allows binding to an scFv-GFP
                     fusion protein (Tanenbaum et al., 2014)"
                     /translation="LLSKNYHLENEVARLKKLVGER"
     CDS             complement(2497..2535)
                     /codon_start=1
                     /product="expressed peptide start"
                     /label=expressed peptide start
                     /protein_id="AAD50824.1"
                     /translation="MSTDRSRRFTTS"
     RBS             complement(2543..2565)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(2597..2615)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3386..3574
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    3679..3821
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4007..4595)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4769..5626)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5627..5731)
                     /label=AmpR promoter
ORIGIN
        1 aattctcatg tttgacagct tatcatcgat aagctttaat gcggtagttt atcacagtta
       61 aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat cgtcatcctc
      121 ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact gccgggcctc
      181 ttgcgggata tcgtccattc cgacagcatc gccagtcact atggcgtgct gctagcgcta
      241 tatgcgttga tgcaatttct atgcgcaccc gttctcggag cactgtccga ccgctttggc
      301 cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc gatcatggcg
      361 accacacccg tcctgtggat atccgggcat atagttcctc ctttcagcaa aaaacccctc
      421 aagacccgtt tagaggcccc aaggggttat gctagttatt gctcagcggt ggcagcagcc
      481 aactcagctt cctttcgggc tttgttagca gcgccggatc ctcaacgttc accaactagt
      541 tttttcaggc gcgcaacttc gttctcgagg tggtagtttt tagacaggag ctcgttatcg
      601 ctatgacgga acaggtattc gctggtcact tcgatggttt gcccggataa acggaactgg
      661 aaaaactgct gctggtgttt tgcttccgtc agcgctggat gcggcgtgcg gtcggcaaag
      721 accagaccgt tcatacagaa ctggcgatcg ttcggcgtat cgccaaaatc accgccgtaa
      781 gccgaccacg ggttgccgtt ttcatcatat ttaatcagcg actgatccac ccagtcccag
      841 acgaagccgc cctgtaaacg gggatactga cgaaacgcct gccagtattt agcgaaaccg
      901 ccaagactgt tacccatcgc gtgggcgtat tcgcaaagga tcagcgggcg cgtctctcca
      961 ggtagcgaaa gccatttttt gatggaccat ttcggcacag ccgggaaggg ctggtcttca
     1021 tccacgcgcg cgtacatcgg gcaaataata tcggtggccg tggtgtcggc tccgccgcct
     1081 tcatactgca ccgggcggga aggatcgaca gatttgatcc agcgatacag cgcgtcgtga
     1141 ttagcgccgt ggcctgattc attccccagc gaccagatga tcacactcgg gtgattacga
     1201 tcgcgctgca ccattcgcgt tacgcgttcg ctcatcgccg gtagccagcg cggatcatcg
     1261 gtcagacgat tcattggcac catgccgtgg gtttcaatat tggcttcatc caccacatac
     1321 aggccgtagc ggtcgcacag cgtgtaccac agcggatggt tcggataatg cgaacagcgc
     1381 acggcgttaa agttgttctg cttcatcagc aggatatcct gcaccatcgt ctgctcatcc
     1441 atgacctgac catgcagagg atgatgctcg tgacggttaa cgcctcgaat cagcaacggc
     1501 ttgccgttca gcagcagcag accattttca atccgcacct cgcggaaacc gacatcgcag
     1561 gcttctgctt caatcagcgt gccgtcggcg gtgtgcagtt caaccaccgc acgatagaga
     1621 ttcgggattt cggcgctcca cagtttcggg ttttcgacgt tcagacgtag tgtgacgcga
     1681 tcggcataac caccacgctc atcgataatt tcaccgccga aaggcgcggt gccgctggcg
     1741 acctgcgttt caccctgcca taaagaaact gttacccgta ggtagtcacg caactcgccg
     1801 cacatctgaa cttcagcctc cagtacagcg cggctgaaat catcattaaa gcgagtggca
     1861 acatggaaat cgctgatttg tgtagtcggt ttatgcagca acgagacgtc acggaaaatg
     1921 ccgctcatcc gccacatatc ctgatcttcc agataactgc cgtcactcca acgcagcacc
     1981 atcaccgcga ggcggttttc tccggcgcgt aaaaatgcgc tcaggtcaaa ttcagacggc
     2041 aaacgactgt cctggccgta accgacccag cgcccgttgc accacagatg aaacgccgag
     2101 ttaacgccat caaaaataat tcgcgtctgg ccttcctgta gccagctttc atcaacatta
     2161 aatgtgagcg agtaacaacc cgtcggattc tccgtgggaa caaacggcgg attgaccgta
     2221 atgggatagg ttacgttggt gtagatgggc gcatcgtaac cgtgcatctg ccagtttgag
     2281 gggacgacga cagtatcggc ctcaggaaga tcgcactcca gccagctttc cggcaccgct
     2341 tctggtgccg gaaaccaggc aaagcgccat tcgccattca ggctgcgcaa ctgttgggaa
     2401 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     2461 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggga
     2521 tcgatccgtc gacatatgta tatctccttc ttaaagttaa acaaaattat ttctagaggg
     2581 aaaccgttgt ggtctcccta tagtgagtcg tattaatttc gcgggatcga gatctcgatc
     2641 ctctacgccg gacgcatcgt ggccggcatc accggcgccg ccctatacct tgtctgcctc
     2701 cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga agccggcggc
     2761 acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg cggagaactg
     2821 tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc tccagcagcc
     2881 gcacgcggcg catctcgggc agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc
     2941 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
     3001 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
     3061 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
     3121 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
     3181 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
     3241 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
     3301 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
     3361 attccccctt acacggaggc atcaagtgac caaacaggaa aaaaccgccc ttaacatggc
     3421 ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga
     3481 tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg
     3541 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     3601 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     3661 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     3721 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat atgcggtgtg
     3781 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     3841 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3901 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3961 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4021 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4081 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4141 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4201 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4261 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4321 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4381 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4441 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4501 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4561 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4621 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4681 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4741 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4801 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4861 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4921 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4981 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5041 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac
     5101 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5161 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5221 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5281 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5341 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc
     5401 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5461 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5521 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5581 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5641 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5701 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5761 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5821 ttcgtcttca ag
//
