LOCUS       40924_26106        5141 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJH370, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5141)
  AUTHORS   Hu JC, O'Shea EK, Kim PS, Sauer RT.
  TITLE     Sequence requirements for coiled-coils: analysis with lambda 
            repressor-GCN4 leucine zipper fusions
  JOURNAL   Science 250 (4986), 1400-1403 (1990)
  PUBMED    2147779
REFERENCE   2  (bases 1 to 5141)
  AUTHORS   Hu JC.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 5141)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5141)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science"; 
            date: "1990"; volume: "250"; issue: "4986"; pages: "1400-1403"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-OCT-2000) Biochemistry "
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..58
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    66..82
                     /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      91..94
                     /regulatory_class="ribosome_binding_site"
     CDS             535..600
                     /label=GCN4_v1
                     /note="GCN4 peptide that allows binding to an scFv-GFP
                     fusion protein (Tanenbaum et al., 2014)"
     terminator      670..717
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             2202..2390
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    2495..2635
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2821..3409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3520..4033
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(4078..4935)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4936..5040)
                     /label=AmpR promoter
ORIGIN
        1 aattctcact cattaggcac cccaggcttt acactttatg cttccggctc gtataatgtg
       61 tggaattgtg agcggataac aatttcacac aggaaacagc gtatgagcac aaaaaagaaa
      121 ccattaacac aagagcagct tgaggacgca cgtcgcctta aagcaattta tgaaaaaaag
      181 aaaaatgaac ttggcttatc ccaggaatct gtcgcagaca agatggggat ggggcagtca
      241 ggcgttggtg ctttatttaa tggcatcaat gcattaaatg cttataacgc cgcattgctt
      301 gcaaaaattc tcaaagttag cgttgaagaa tttagccctt caatcgccag agaaatctac
      361 gagatgtatg aagcggttag tatgcagccg tcacttagaa gtgagtatga gtaccctgtt
      421 ttttctcatg ttcaggcagg gatgttctca cctaagctta gaacctttac caaaggtgat
      481 gcggagagat gggtgtcgac acatatgaaa cagctggaag acaaagttga agagctcctg
      541 tctaaaaact accacctcga gaacgaagtt gcgcgcctga aaaaactagt tggtgaacgt
      601 tgaggatccg gcgctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
      661 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
      721 aaaggaggaa ctatatgccc ggatatcgtc cattccgaca gcatcgccag tcactatggc
      781 gtgctgctag cgctatatgc gttgatgcaa tttctatgcg cacccgttct cggagcactg
      841 tccgaccgct ttggccgccg cccagtcctg ctcgcttcgc tacttggagc cactatcgac
      901 tacgcgatca tggcgaccac acccgtcctg tggatctcga ccgatgccct tgagagcctt
      961 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     1021 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg
     1081 cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat
     1141 cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa
     1201 gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc
     1261 gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat
     1321 gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca
     1381 aggatcgctc gcggctctta ccagcctaac ttcgatcact ggaccgctga tcgtcacggc
     1441 gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag gcgccgccct
     1501 ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg
     1561 aatggaagcc ggcggcacct cgctaacgga ttcaccactc caagaattgg agccaatcaa
     1621 ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc agaacatatc catcgcgtcc
     1681 gccatctcca gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg
     1741 cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt
     1801 tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct
     1861 gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg
     1921 cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta
     1981 ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt
     2041 ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg
     2101 gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt
     2161 acccccatga acagaaattc ccccttacac ggaggcatca agtgaccaaa caggaaaaaa
     2221 ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca
     2281 acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg
     2341 agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2401 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2461 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     2521 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     2581 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
     2641 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2701 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2761 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2821 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2881 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2941 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3001 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3061 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3121 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3181 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3241 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     3301 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3361 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3421 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3481 catgagatta tcaaaaagga tcttcaccta gatcctttta cgcgccctgt agcggcgcat
     3541 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3601 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3661 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3721 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3781 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3841 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     3901 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3961 taacgtttac aatttaaata tttgcttata caatcttcct gtttttgggg cttttctgat
     4021 tatcaaccgg ggtaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4081 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4141 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4201 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4261 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4321 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4381 tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4441 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4501 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4561 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4621 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4681 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4741 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4801 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4861 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4921 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4981 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5041 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5101 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa g
//
