LOCUS       40924_28726        5070 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLS3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5070)
  AUTHORS   Hays LB, Chen Y-S.A., Hu JC.
  TITLE     A two-hybrid system for characterization of protein-protein 
            interactions in E. coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5070)
  AUTHORS   Hays LB, Chen Y-S.A., Hu JC.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 5070)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5070)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-JUL-1999) Biochemistry "
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5070
                     /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)."
     CDS             109..525
                     /codon_start=1
                     /product="434-GCN4-linker"
                     /label=434-GCN4-linker
                     /note="fusion protein; contains N-terminal DNA binding
                     domain from bacteriophage 434 repressor, a modified leucine
                     zipper from GCN4 and a linker for fusion to additional 
                     protein domains"
                     /protein_id="AAD50896.1"
                     /translation="MSISSRVKSKRIQLGLNQAELAQKVGTTQQSIEQLENGKTKRPRF
                     LPELASALGVSVDWLLNGTSDSNVRFVGHVEPKGKKLRTFTKGDAERWVSTHMKQLEDK
                     IEELLSKNYHLENEIARLKKLIGERGGQLNASGS"
     terminator      599..646
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             2131..2319
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    2424..2564
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2750..3338)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3449..3962
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(4007..4864)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4865..4969)
                     /label=AmpR promoter
ORIGIN
        1 aattctcact cattaggcac cccaggcttt acactttatg cttccggctc gtataatgtg
       61 tggaattgtg agcggataac aatttcacat ctagacagga aacagcttat gagtatttct
      121 tccagggtaa aaagcaaaag aattcagctt ggacttaacc aggctgaact tgctcaaaag
      181 gtggggacta cccagcagtc gatcgagcag ctcgaaaacg gtaaaactaa gcgaccacgc
      241 tttttaccag aacttgcgtc agctcttggc gtaagtgttg actggctgct caatggcacc
      301 tctgattcga atgttagatt tgttgggcac gttgagccca aagggaaaaa gcttagaacc
      361 tttaccaaag gtgatgcgga gagatgggtg tcgacacata tgaaacagct ggaagacaaa
      421 atcgaagagc tcctgtctaa aaactaccac ctcgagaacg aaatcgcgcg cctgaaaaaa
      481 ctaatcggtg aacgtggtgg ccaattgaac gcgtcgggat cctaagtaac taagatccgg
      541 cgctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
      601 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
      661 tatatgcccg gatatcgtcc attccgacag catcgccagt cactatggcg tgctgctagc
      721 gctatatgcg ttgatgcaat ttctatgcgc acccgttctc ggagcactgt ccgaccgctt
      781 tggccgccgc ccagtcctgc tcgcttcgct acttggagcc actatcgact acgcgatcat
      841 ggcgaccaca cccgtcctgt ggatctcgac cgatgccctt gagagccttc aacccagtca
      901 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
      961 tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct
     1021 ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc
     1081 tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta
     1141 tcgccggcat ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct
     1201 ggatggcctt ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc
     1261 aggccatgct gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg
     1321 cggctcttac cagcctaact tcgatcactg gaccgctgat cgtcacggcg atttatgccg
     1381 cctcggcgag cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct
     1441 gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg
     1501 gcggcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag
     1561 aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag
     1621 cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt
     1681 gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga
     1741 atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc
     1801 aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc
     1861 gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac
     1921 acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg
     1981 ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc
     2041 atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa
     2101 cagaaattcc cccttacacg gaggcatcaa gtgaccaaac aggaaaaaac cgcccttaac
     2161 atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac
     2221 gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc
     2281 agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     2341 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     2401 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
     2461 tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     2521 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct
     2581 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2641 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2701 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2761 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2821 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2881 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2941 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3001 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3061 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3121 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3181 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3241 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3301 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3361 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3421 caaaaaggat cttcacctag atccttttac gcgccctgta gcggcgcatt aagcgcggcg
     3481 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3541 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3601 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3661 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3721 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3781 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3841 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     3901 atttaaatat ttgcttatac aatcttcctg tttttggggc ttttctgatt atcaaccggg
     3961 gtaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4021 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4081 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4141 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4201 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4261 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg
     4321 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4381 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4441 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4501 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4561 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4621 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4681 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4741 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4801 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4861 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4921 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4981 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5041 ggcgtatcac gaggcccttt cgtcttcaag
//
