LOCUS       62056_18955        3773 bp DNA     circular SYN 10-APR-2019
DEFINITION  Cloning vector pRNBZMB, complete sequence.
ACCESSION   MH507507
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3773)
  AUTHORS   Kumar D, Batra J, Komives C, Rathore AS.
  TITLE     QbD Based Media Development for the Production of Fab Fragments in 
            E. coli
  JOURNAL   Bioengineering (Basel) 6 (2), E29 (2019)
  PUBMED    30925730
REFERENCE   2  (bases 1 to 3773)
  AUTHORS   Kumar D, Batra J, Komives C, Rathore AS.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JUN-2018) Chemical Engineering, Indian Institute of 
            Technology Delhi, Lab-94, Block-II, Hauz Khas, New Delhi, Delhi 
            110016, India
REFERENCE   3  (bases 1 to 3773)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Bioengineering (Basel)"; date: "2019"; volume: "6"; issue: "2"; 
            pages: "E29"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-JUN-2018) Chemical Engineering, Indian Institute of Technology 
            Delhi, Lab-94, Block-II, Hauz Khas, New Delhi, Delhi 110016, India"
FEATURES             Location/Qualifiers
     source          1..3773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..774
                     /codon_start=1
                     /transl_table=11
                     /product="Mal/Ranibizumab heavy chain"
                     /label=Mal/Ranibizumab heavy chain
                     /note="optimized for E. coli. expression"
                     /protein_id="QBW95968.1"
                     /translation="MKIKTGARILALSALTTMMFSASALAEVQLVESGGGLVQPGGSLR
                     LSCAASGYDFTHYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKST
                     AYLQMNSLRAEDTAVYYCAKYPYYYGTSHWYFDVWGQGTLVTVSSASTKGPSVFPLAPS
                     SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS
                     SSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHL"
     sig_peptide     813..878
                     /label=pelB signal sequence
                     /note="leader peptide for secretion"
     CDS             1203..1520
                     /label=hIg-kappa-CL
                     /note="Human immunoglobulin kappa light chain constant
                     region"
     terminator      1555..1602
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1700..1729
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1759..2304
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     terminator      complement(2569..2596)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2623..3429)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(3430..3520)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     promoter        3624..3742
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
ORIGIN
        1 atgaaaataa aaacaggtgc acgcatcctc gcattatccg cattaacgac gatgatgttt
       61 tccgcctcgg ctctcgccga agtccaactg gtcgagagcg gtggcggcct ggtgcagccg
      121 ggtggcagcc tgcgcctgag ctgcgccgca agcggctatg attttacgca ttacggcatg
      181 aactgggttc gccaggctcc gggcaaaggt ctggaatggg ttggctggat caatacctac
      241 accggtgagc caacgtacgc cgcagacttc aagcgtcgtt tcaccttttc actggatacc
      301 agcaagagca cggcctactt gcaaatgaac tcgctgcgtg cagaggacac tgcggtgtat
      361 tactgtgcga agtatcctta ctattatggt acgagccact ggtacttcga tgtctggggt
      421 cagggtactc tggttacggt gagcagcgcg agcaccaaag gtccgtccgt gttcccgctg
      481 gcgccgagct ccaagagcac cagcggtggc accgctgcgc tgggttgtct ggtgaaagac
      541 tattttccgg aaccggtgac cgttagctgg aatagcggtg cgctgacgtc cggtgttcac
      601 acgttcccgg cagtgctgca atctagcggc ctgtactccc tgagcagcgt cgtcaccgtt
      661 ccgagctcta gcttgggtac ccagacgtat atctgcaatg tcaaccacaa gccgagcaac
      721 accaaggttg acaaaaaagt cgagccaaaa agctgcgaca aaacccattt gtgataacac
      781 tgtataacat taagatttta aggaggtaaa aaatgaaata cctgctgccg accgctgctg
      841 ctggtctgct gctcctcgct gcccagccgg cgatggccga catccaactg acccagagcc
      901 cgtctagcct gtctgcgagc gtgggtgacc gtgttaccat tacgtgcagc gctagccaag
      961 acattagcaa ctatctgaat tggtaccagc agaaaccggg caaggcccct aaggtcctga
     1021 tctactttac cagctccctg cacagcggtg taccgtcccg cttctcgggc agcggctcgg
     1081 gcaccgactt caccctgacg attagcagcc tgcagccaga ggattttgcg acttactatt
     1141 gtcaacagta cagcaccgtc ccgtggactt tcggtcaagg taccaaagtc gagattaagc
     1201 gcaccgtggc ggcaccgagc gttttcatct ttccgccgag cgatgaacaa ctgaagtctg
     1261 gtacggcgag cgttgtttgt ttgttgaaca acttttaccc gcgtgaggca aaagtgcagt
     1321 ggaaagttga caatgcgctg cagagcggta actctcaaga gagcgtgacg gaacaggata
     1381 gcaaagattc cacctatagc ctgtccagca ccctgacgct gagcaaagcc gattatgaga
     1441 agcacaaagt gtacgcgtgc gaagtcacgc atcaaggtct gagcagcccg gtgaccaagt
     1501 cattcaatcg tggcgaatgc taaggttaga gcggccgcca ccgctgagca ataactagca
     1561 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     1621 tccgggtaac gaattcaagc ttgatatcat tcaggacgag cctcagactc cagcgtaact
     1681 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaagtctt
     1741 cttaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa
     1801 cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa
     1861 ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta
     1921 accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg
     1981 gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc
     2041 ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg
     2101 aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta
     2161 aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa cgcctggtat
     2221 ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg
     2281 tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa
     2341 gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc
     2401 gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga agcggaatat atcctgtatc
     2461 acatattctg ctgacgcacc ggtgcagcct tttttctcct gccacatgaa gcacttcact
     2521 gacaccctca tcagtgccaa catagtaagc cagtatacac tccgctagcg cagaaaggcc
     2581 cacccgaagg tgagccaggt gattacattt gggccctcat tagaaaaact catcgagcat
     2641 caaatgaaat tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     2701 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     2761 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     2821 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     2881 aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     2941 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga ggcgaaatac
     3001 gcgatcgctg ttaaaaggac aattacaaac aggaatcgag tgcaaccggc gcaggaacac
     3061 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaacgc
     3121 tgtttttccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     3181 cttgatggtc ggaagtggca taaattccgt cagccagttt agtctgacca tctcatctgt
     3241 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     3301 cccatacaag cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     3361 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgacgttt cccgttgaat
     3421 atggctcata gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     3481 ttcattatgg tgaaagttgg aacctcttac gtgccgatca agaagacggt caaaagcctc
     3541 cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagtatt
     3601 gagcgatatc tagagaattc gtccaccaca attcagcaaa ttgtgaacat catcacgttc
     3661 atctttccct ggttgccaat ggcccatttt cctgtcagta acgagaaggt cgcgaattca
     3721 ggcgcttttt agactggtcg taatgaaatt ctttttaaga aggagatata cat
//
