LOCUS       Exported                4041 bp ds-DNA     circular SYN 23-FEB-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4041)
  AUTHORS   gep
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Feb 25, 2020 from SnapGene Viewer 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4041
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1061
                     /codon_start=1
                     /label=AmpR
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
     protein_bind    2108..2129
                     /label=CAP binding site
     promoter        2144..2174
                     /label=lac promoter
     protein_bind    2182..2198
                     /label=lac operator
     primer_bind     2206..2222
                     /label=M13 rev
     CDS             2218..3896
                     /codon_start=1
                     /label=lacZ-alpha
                     /translation="MTMITPSFVDTTIK**LHP*IDNYHTHKLILPLHGLNFSKFTTFS
                     *RGHFRGSGVEDRDLDPAKLIRLTIGEL*ADNNSPLEIILFNFKKEIYICIRI*KEKSS
                     LLQELLQGSERRWPFASARSRQKWLSTIIVINKIRTR*KKRSSRRAVKLLSSKEMSRKR
                     KM*KISCKRQLRSSAHSIL*LIMPVLKILCHLTKCRSRIGIKSSART*RVPF*EAVKRL
                     NIS*KTISREMSLTCPVCTK*FLGHYLSTMRQVKAG*S**HAH*RWNTRRRAFASIILG
                     QVRSTRRSIRRNLLTLNRELM*KA*FQWDISANRRRSPQ*QPGLLRRKPATSQASRYSR
                     TAV*HFILHSRQAAVNDSSTPPPPLLEHHHHHH*DPAANKARKEAELAAATAEQ*LA*P
                     LGASKRVLRGFLLKGGTISGSRVLTAVVLLKYSIHKNINLWLFCEISIYGCFVVKLLII
                     NNY*ISLYGCINRYRARIHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIA
                     LPNSCAA*MANGA*CGIFSLRICAVFHTAYGALSVQSALMPH"
     misc_feature    2235..3634
                     /label=pLR_Cm
     misc_feature    2241..2376
                     /label=RE
     promoter        2411..2429
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2430..2454
                     /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)."
     CDS             2499..3311
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MYPDLKGKVVAITGAASGLGKAMAIRFGKEQAKVVINYYSNKQDP
                     NEVKEEVIKAGGEAVVVQGDVTKEEDVKNIVQTAIKEFGTLDIMINNAGLENPVPSHEM
                     PLKDWDKVIGTNLTGAFLGSREAIKYFVENDIKGNVINMSSVHEVIPWPLFVHYAASKG
                     GMKLMTRTLALEYAPKGIRVNNIGPGAINTTINKEKFADPEQRADVESMIPMGYIGEPE
                     EIAAVAAWLASKEASYVTGITLFADGGMTLYPSFQAGRG**LEHTTTTT"
     CDS             2499..3284
                     /codon_start=1
                     /label=GDH
                     /translation="MYPDLKGKVVAITGAASGLGKAMAIRFGKEQAKVVINYYSNKQDP
                     NEVKEEVIKAGGEAVVVQGDVTKEEDVKNIVQTAIKEFGTLDIMINNAGLENPVPSHEM
                     PLKDWDKVIGTNLTGAFLGSREAIKYFVENDIKGNVINMSSVHEVIPWPLFVHYAASKG
                     GMKLMTRTLALEYAPKGIRVNNIGPGAINTTINKEKFADPEQRADVESMIPMGYIGEPE
                     EIAAVAAWLASKEASYVTGITLFADGGMTLYPSFQAGRG"
     misc_feature    3297..3321
                     /label=缺失
     CDS             3319..3336
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3403..3450
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    3476..3628
                     /label=LE
     misc_feature    3635..3646
                     /label=MCS
     primer_bind     complement(3647..3663)
                     /label=M13 fwd
     primer_bind     complement(3880..3897)
                     /label=Donor-250R
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt tgttgataca accataaaat gataattaca cccataaatt
     2281 gataattatc acacccataa attgatattg cctcttcatg gtctaaactt cagtaagttt
     2341 acgacatttt catgaagagg tcatttccgg ggatccggcg tagaggatcg agatctcgat
     2401 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta
     2461 gaaataattt tgtttaactt taagaaggag atatacatat gtatccggat ttaaaaggaa
     2521 aagtcgtcgc tattacagga gctgcttcag ggctcggaaa ggcgatggcc attcgcttcg
     2581 gcaaggagca ggcaaaagtg gttatcaact attatagtaa taaacaagat ccgaacgagg
     2641 taaaagaaga ggtcatcaag gcgggcggtg aagctgttgt cgtccaagga gatgtcacga
     2701 aagaggaaga tgtaaaaaat atcgtgcaaa cggcaattaa ggagttcggc acactcgata
     2761 ttatgattaa taatgccggt cttgaaaatc ctgtgccatc tcacgaaatg ccgctcaagg
     2821 attgggataa agtcatcggc acgaacttaa cgggtgcctt tttaggaagc cgtgaagcga
     2881 ttaaatattt cgtagaaaac gatatcaagg gaaatgtcat taacatgtcc agtgtgcacg
     2941 aagtgattcc ttggccatta tttgtccact atgcggcaag taaaggcggg atgaagctga
     3001 tgacacgcac attagcgttg gaatacgcgc cgaagggcat tcgcgtcaat aatattgggc
     3061 caggtgcgat caacacgacg atcaataagg agaaatttgc tgaccctgaa cagagagctg
     3121 atgtagaaag catgattcca atgggatata tcggcgaacc ggaggagatc gccgcagtag
     3181 cagcctggct tgcttcgaag gaagccagct acgtcacagg catcacgtta ttcgcggacg
     3241 gcggtatgac actttatcct tcattccagg caggccgcgg ttaatgactc gagcacacca
     3301 ccaccaccac tactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
     3361 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     3421 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatctaga
     3481 gtccttactg cagtagtttt gctgaaatac tcgattcaca aaaatatcaa cttatggttg
     3541 ttttgtgaga tatcaatata tggttgtttt gtggttaagt tgctgattat aaataattat
     3601 taaatatcac tttatggttg catcaacagg taccgagctc gaattcactg gccgtcgttt
     3661 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     3721 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     3781 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     3841 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3901 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3961 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4021 cgtcatcacc gaaacgcgcg a
//
