LOCUS       40924_5234        8342 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Bacterial two-hybrid vector pB2H-delta-omega, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8342)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8342)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 8342)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8342)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     RBS             42..64
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             complement(2268..2441)
                     /codon_start=1
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     misc_feature    complement(2503..2538)
                     /label=5' UTR of lacZ gene, incl. RBS
                     /note="5' UTR of lacZ gene, incl. RBS"
     protein_bind    2520..2536
                     /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)."
     promoter        complement(2544..2572)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     promoter        complement(2601..2619)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2637..2653)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2661..2677)
                     /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        complement(2685..2715)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2730..2751)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3068..3096
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             3292..3336
                     /codon_start=1
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /translation="KETAAAKFERQHMDS"
     terminator      3388..3435
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3472..3927
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(4045..4902)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4903..4995)
                     /label=AmpR promoter
     rep_origin      5076..5664
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(5850..5992)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6097..6285)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6860..7939)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(7940..8017)
                     /label=lacI promoter
     promoter        8326..8342
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atggttatgc acgggtgaac tgatcgcgca gcggcgtcag cagttgtttt
      121 ttatcgccaa tccacatctg tgaaagaaag cctgactggc ggttaaattg ccaacgctta
      181 ttacccagct cgatgcaaaa atccatttcg ctggtggtca gatgcgggat ggcgtgggac
      241 gcggcgggga gcgtcacact gaggttttcc gccagacgcc actgctgcca ggcgctgatg
      301 tgcccggctt ctgaccatgc ggtcgcgttc ggttgcacta cgcgtactgt gagccagagt
      361 tgcccggcgc tctccggctg cggtagttca ggcagttcaa tcaactgttt accttgtgga
      421 gcgacatcca gaggcacttc accgcttgcc agcggcttac catccagcgc caccatccag
      481 tgcaggagct cgttatcgct atgacggaac aggtattcgc tggtcacttc gatggtttgc
      541 ccggataaac ggaactggaa aaactgctgc tggtgttttg cttccgtcag cgctggatgc
      601 ggcgtgcggt cggcaaagac cagaccgttc atacagaact ggcgatcgtt cggcgtatcg
      661 ccaaaatcac cgccgtaagc cgaccacggg ttgccgtttt catcatattt aatcagcgac
      721 tgatccaccc agtcccagac gaagccgccc tgtaaacggg gatactgacg aaacgcctgc
      781 cagtatttag cgaaaccgcc aagactgtta cccatcgcgt gggcgtattc gcaaaggatc
      841 agcgggcgcg tctctccagg tagcgaaagc cattttttga tggaccattt cggcacagcc
      901 gggaagggct ggtcttcatc cacgcgcgcg tacatcgggc aaataatatc ggtggccgtg
      961 gtgtcggctc cgccgccttc atactgcacc gggcgggaag gatcgacaga tttgatccag
     1021 cgatacagcg cgtcgtgatt agcgccgtgg cctgattcat tccccagcga ccagatgatc
     1081 acactcgggt gattacgatc gcgctgcacc attcgcgtta cgcgttcgct catcgccggt
     1141 agccagcgcg gatcatcggt cagacgattc attggcacca tgccgtgggt ttcaatattg
     1201 gcttcatcca ccacatacag gccgtagcgg tcgcacagcg tgtaccacag cggatggttc
     1261 ggataatgcg aacagcgcac ggcgttaaag ttgttctgct tcatcagcag gatatcctgc
     1321 accatcgtct gctcatccat gacctgacca tgcagaggat gatgctcgtg acggttaacg
     1381 cctcgaatca gcaacggctt gccgttcagc agcagcagac cattttcaat ccgcacctcg
     1441 cggaaaccga catcgcaggc ttctgcttca atcagcgtgc cgtcggcggt gtgcagttca
     1501 accaccgcac gatagagatt cgggatttcg gcgctccaca gtttcgggtt ttcgacgttc
     1561 agacgtagtg tgacgcgatc ggcataacca ccacgctcat cgataatttc accgccgaaa
     1621 ggcgcggtgc cgctggcgac ctgcgtttca ccctgccata aagaaactgt tacccgtagg
     1681 tagtcacgca actcgccgca catctgaact tcagcctcca gtacagcgcg gctgaaatca
     1741 tcattaaagc gagtggcaac atggaaatcg ctgatttgtg tagtcggttt atgcagcaac
     1801 gagacgtcac ggaaaatgcc gctcatccgc cacatatcct gatcttccag ataactgccg
     1861 tcactccaac gcagcaccat caccgcgagg cggttttctc cggcgcgtaa aaatgcgctc
     1921 aggtcaaatt cagacggcaa acgactgtcc tggccgtaac cgacccagcg cccgttgcac
     1981 cacagatgaa acgccgagtt aacgccatca aaaataattc gcgtctggcc ttcctgtagc
     2041 cagctttcat caacattaaa tgtgagcgag taacaacccg tcggattctc cgtgggaaca
     2101 aacggcggat tgaccgtaat gggataggtt acgttggtgt agatgggcgc atcgtaaccg
     2161 tgcatctgcc agtttgaggg gacgacgaca gtatcggcct caggaagatc gcactccagc
     2221 cagctttccg gcaccgcttc tggtgccgga aaccaggcaa agcgccattc gccattcagg
     2281 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     2341 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     2401 cgttgtaaaa cgacggccag tgaatccgta atcatggtca tacccccgga tccgaattcg
     2461 agctcggcgc gcctgcagcg gccgcatgca tatggatcga tcctgtttcc tgtgtgaaat
     2521 tgttatccgc tcacaattcc acacattata cgagccgatg attaattgtc aacaggggga
     2581 tggggagtaa gcttgagtat tctatagtgt cacctaaata gcttggcgta atcatggtca
     2641 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2701 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2761 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2821 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2881 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2941 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgaattaatt ctcatgtttg
     3001 acagcttatc atcggataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtt
     3061 ctcatgtttg acagcttatc ttcgattagc tttaatgcgg tagtttatca cagttaaatt
     3121 gctaacgcag tcaggcaccg tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca
     3181 ccgtcaccct ggatgctgta ggcataggct tggttatgcc ggtactgccg ggcctcttgc
     3241 gggatatcgg ccggccacgc gatcgctgac gtcggtaccc tcgagtctgg taaagaaacc
     3301 gctgctgcga aatttgaacg ccagcacatg gactcgtcta ctagcgcagc ttaattaacc
     3361 taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     3421 ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct
     3481 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3541 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3601 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3661 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3721 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3781 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3841 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3901 ttaacaaaat attaacgttt acaatttctg gcggcacgat ggcatgagat tatcaaaaag
     3961 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4021 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4081 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4141 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4201 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4261 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4321 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4381 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4441 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4501 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4561 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4621 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4681 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4741 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4801 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4861 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaatc
     4921 atgattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4981 agaaaaataa acaaataggt catgaccaaa atcccttaac gtgagttttc gttccactga
     5041 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5101 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5161 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5221 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5281 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5341 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5401 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5461 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5521 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5581 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5641 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5701 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5761 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5821 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     5881 tgcggtattt cacaccgcat atatggtgca ctctcagtac aatctgctct gatgccgcat
     5941 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     6001 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     6061 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     6121 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     6181 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     6241 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     6301 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     6361 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     6421 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     6481 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
     6541 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     6601 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     6661 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     6721 caacgacagg agcacgatca tgctagtcat gccccgcgcc caccggaagg agctgactgg
     6781 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     6841 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6901 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     6961 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     7021 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     7081 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt
     7141 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     7201 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     7261 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     7321 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     7381 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     7441 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     7501 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     7561 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     7621 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     7681 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     7741 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     7801 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     7861 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     7921 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     7981 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     8041 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     8101 gcaaggaatg gtgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     8161 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     8221 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     8281 tgcgtccggc gtagaggatc gagatcgatc tcgatcccgc gaaattaata cgactcacta
     8341 ta
//
