LOCUS       40924_18216        5434 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pET-mod, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5434)
  AUTHORS   Soleyman M, Ebrahimi M, Khalili M.
  TITLE     Modification of pET expression vector for ELPylation of target 
            protein for construct of functional active biomaterial
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5434)
  AUTHORS   Soleyman M, Ebrahimi M, Khalili M.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-FEB-2015) Medical Biotechnology, Arak Medical Science 
            University, Arak, Markazi 0098, Iran
REFERENCE   3  (bases 1 to 5434)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5434)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-FEB-2015) Medical Biotechnology, Arak Medical Science 
            University, Arak, Markazi 0098, Iran"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(8..32)
                     /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(33..51)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        364..441
                     /label=lacI promoter
     CDS             442..1521
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    1537..1558
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2333..2521
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    2626..2768
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2954..3542)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3716..4573)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4574..4678)
                     /label=AmpR promoter
     rep_origin      complement(4705..5160)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     terminator      complement(5197..5244)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5311..5328)
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     RBS             complement(5405..5427)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ctagagggga attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg
       61 gatcgagatc gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
      121 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
      181 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
      241 gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc
      301 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc
      361 ccggacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag
      421 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
      481 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
      541 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
      601 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
      661 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
      721 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
      781 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
      841 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
      901 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
      961 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     1021 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     1081 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     1141 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     1201 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac
     1261 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     1321 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     1381 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     1441 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     1501 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
     1561 ggcaccggga tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     1621 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     1681 aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc
     1741 gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt
     1801 cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc
     1861 ggccccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     1921 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     1981 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     2041 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     2101 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     2161 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     2221 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     2281 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     2341 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     2401 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     2461 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     2521 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     2581 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     2641 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     2701 ctgagagtgc accatatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     2761 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2821 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2881 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2941 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3001 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3061 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3121 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3181 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3241 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3301 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3361 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     3421 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3481 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3541 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3601 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3661 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3721 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3781 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3841 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3901 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3961 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4021 ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4081 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4141 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4201 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4261 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4321 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4381 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4441 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4501 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4561 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4621 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4681 catttccccg aaaagtgcca cctgaaattg taaacgttaa tattttgtta aaattcgcgt
     4741 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4801 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4861 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4921 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4981 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     5041 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     5101 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     5161 cccattcgcc aatccggata tagttcctcc tttcagcaaa aaacccctca agacccgttt
     5221 agaggcccca aggggttatg ctagttattg ctcagcggtg gcagcagcca actcagcttc
     5281 ctttcgggct ttgttagcag ccggatctca gtggtggtgg tggtggtgct cgagccagta
     5341 ccagtaccac ggccagcccg gcccagaacc accagaacca ccgaattcgg atcccatatg
     5401 tatatctcct tcttaaagtt aaacaaaatt attt
//
