LOCUS       40924_5164        3809 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAZ1LT7, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3809)
  AUTHORS   Guggenbichler F, Buttner C, Rudolph W, Zimmermann K, Gunzer F, 
            Pohlmann C.
  TITLE     Design of a covalently linked human interleukin-10 fusion protein 
            and its secretory expression in Escherichia coli
  JOURNAL   Appl. Microbiol. Biotechnol. (2016) In press
  PUBMED    27430741
REFERENCE   2  (bases 1 to 3809)
  AUTHORS   Guggenbichler F, Buettner C, Rudolph W, Zimmermann K, Gunzer F, 
            Poehlmann C.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JAN-2016) Institute of Medical Microbiology and 
            Hygiene, TU Dresden, Fetscherstrasse 74, Dresden, Saxony 01307, 
            Germany
REFERENCE   3  (bases 1 to 3809)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3809)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Microbiol. Biotechnol. (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-JAN-2016) Institute of Medical Microbiology and Hygiene, TU 
            Dresden, Fetscherstrasse 74, Dresden, Saxony 01307, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             457..479
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     gene            487..1563
                     /gene="hly-10LT7"
                     /label=hly-10LT7
     CDS             487..1560
                     /codon_start=1
                     /gene="hly-10LT7"
                     /product="interleukin-10/OmpF fusion protein"
                     /label=hly-10LT7
                     /note="human interleukin-10 fusion protein with ompF
                     leader"
                     /protein_id="ANV82811.1"
                     /translation="MMKRNILAVIVPALLVAGTANASPGQGTQSENSCTHFPGNLPNML
                     RDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAEN
                     QDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMS
                     EFDIFINYIEAYMTMKIRNGGGGSGGGGSGGGGSSPGQGTQSENSCTHFPGNLPNMLRD
                     LRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQD
                     PDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEF
                     DIFINYIEAYMTMKIRN"
     primer_bind     complement(1588..1604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1612..1628)
                     /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(1636..1666)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1681..1702)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1990..2578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2752..3609)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3610..3714)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctctaa tacgaccact
      421 atagggagac cacaacggtt tccctctaga aataattttg tttaacttta agaaggagat
      481 atacatatga tgaagcgcaa tattctggca gtgatcgtcc ctgctctgtt agtagcaggt
      541 actgcaaacg ctagcccagg ccaaggtact cagtctgaaa actcctgtac ccactttccg
      601 ggcaacctgc cgaacatgct gcgtgacctg cgtgatgcgt ttagccgtgt taagaccttc
      661 ttccagatga aagaccagct ggataatctg ctgctgaaag aatctctgct ggaagatttc
      721 aaaggttatc tgggctgcca ggcactgtcc gaaatgatcc agttctacct ggaggaagta
      781 atgccgcagg cggagaacca ggacccggat atcaaagctc atgttaactc cctgggtgaa
      841 aacctgaaaa cgctgcgcct gcgcctgcgt cgttgtcacc gcttcctgcc ttgcgaaaac
      901 aaatctaaag cagtggaaca agtcaagaac gctttcaaca aactgcagga aaaaggtatc
      961 tataaagcga tgtctgagtt tgacatcttc attaattaca ttgaagccta catgaccatg
     1021 aagatccgta acggtggtgg tggtagtggt ggcggtggtt caggtggtgg tggcagcagc
     1081 ccaggccaag gtactcagtc tgaaaactcc tgtacccact ttccgggcaa cctgccgaac
     1141 atgctgcgtg acctgcgtga tgcgtttagc cgtgttaaga ccttcttcca gatgaaagac
     1201 cagctggata atctgctgct gaaagaatct ctgctggaag atttcaaagg ttatctgggc
     1261 tgccaggcac tgtccgaaat gatccagttc tacctggagg aagtaatgcc gcaggcggag
     1321 aaccaggacc cggatatcaa agctcatgtt aactccctgg gtgaaaacct gaaaacgctg
     1381 cgcctgcgcc tgcgtcgttg tcaccgcttc ctgccttgcg aaaacaaatc taaagcagtg
     1441 gaacaagtca agaacgcttt caacaaactg caggaaaaag gtatctataa agcgatgtct
     1501 gagtttgaca tcttcattaa ttacattgaa gcctacatga ccatgaagat ccgtaactaa
     1561 taagcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     1621 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     1681 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     1741 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1801 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1861 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1921 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1981 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2041 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2101 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2161 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2221 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2281 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2341 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2401 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2461 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2521 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2581 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2641 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2701 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2761 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2821 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2881 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2941 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3001 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3061 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3121 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3181 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3241 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3301 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3361 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3421 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3481 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3541 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3601 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3661 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3721 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     3781 aaataggcgt atcacgaggc cctttcgtc
//
