LOCUS       pBAD-MTX_Snifit         5768 bp ds-DNA     circular SYN 15-OCT-2018
DEFINITION  pBAD-SNAP_Pro30_CLIP_DHFR withR44L and H45Q.
ACCESSION
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid.
REFERENCE   1  (bases 1 to 5768)
  AUTHORS   QQQ
  TITLE     Direct Submission
  JOURNAL   Exported Oct 15, 2018 from SnapGene 4.1.6 http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5768
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    224..243
                     /standard_name="pBAD-FP GATC"
     promoter        264..292
                     /gene="ARA prom"
                     /note="Promoter"
     CDS             344..367
                     /codon_start=1
                     /translation="WSHPQFEK"
                     /standard_name="Strep II"
     CDS             374..388
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /translation="DDDDK"
                     /standard_name="enterokinase site"
     CDS             398..949
                     /codon_start=1
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIIFLGKGTSAADAV
                     EVPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKL
                     LKVVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEG
                     GLAVKEWLLAHEGHRLGKPGLGEF"
                     /standard_name="SNAP"
     CDS             950..1039
                     /codon_start=1
                     /translation="PPPPPPPPPPPPPPPPPPPPPPPPPPPPPP"
                     /standard_name="Pro30"
     CDS             1049..1594
                     /codon_start=1
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHKIIFLGKGTSAADAV
                     EVPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKL
                     LKVVKFGEVISESHLAALVGNPAATAAVNTALDGNPVPILIPCHRVVQGDSDVGPYLG
                     GLAVKEWLLAHEGHRLGKPGLG"
                     /standard_name="CLIP"
     CDS             1601..2077
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /translation="MISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGL
                     QTWESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQ
                     FLPKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
                     /standard_name="ecDHFR"
     CDS             2108..2137
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /translation="HHHHHHHHHH"
                     /standard_name="10xHis"
     terminator      2234..2391
                     /gene="rrnB term"
                     /note="Terminator"
     terminator      2357..2400
                     /gene="rrnB_T1 term"
                     /note="Terminator"
     terminator      2532..2559
                     /gene="rrnB_T2 term"
                     /note="Terminator"
     promoter        2601..2629
                     /gene="amp prom"
                     /note="Promoter"
     CDS             2671..3531
                     /codon_start=1
                     /note="ampR"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /standard_name="ampR"
     rep_origin      3686..4305
     CDS             complement(4880..5767)
                     /codon_start=1
                     /translation="MKSMAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLG
                     MKGYILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFR
                     PRAYWHEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLE
                     QLLLRRMEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLF
                     RQQLGISVLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASP
                     SEFRAGCEEKVNDVAVKLS"
                     /standard_name="AraC"
ORIGIN      
        1 ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc
       61 gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg
      121 taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc
      181 agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta
      241 tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca
      301 tacccgtttt ttgggctaac aggaggaatt aaccatggca agctggagcc acccgcagtt
      361 cgaaaagggt gcagatgacg acgacaaggt accgcatatg gataaagact gcgaaatgaa
      421 acgcactacc ctggacagcc cgctgggtaa actggaactg agcggttgcg agcagggcct
      481 gcacgaaatc attttcctgg gcaaaggtac gagcgctgcg gatgcggtcg aagtaccggc
      541 tccggctgcg gttctgggtg gcccggagcc gctgatgcag gctaccgcat ggctgaacgc
      601 atattttcac cagccagagg cgattgaaga gttccctgta ccggccctgc atcacccggt
      661 atttcagcag gaatccttta cccgtcaggt actgtggaaa ctgctgaaag tagtgaaatt
      721 cggcgaggtg atctcctatt cccatctggc ggcgctggct ggcaacccag cggcaactgc
      781 ggcggtgaaa accgctctgt ctggtaatcc ggtgccaatt ctgatcccgt gccaccgtgt
      841 ggttcagggc gacctggacg taggtggcta cgaaggcggc ctggctgtga aagaatggct
      901 gctggcgcac gaaggtcacc gtctgggtaa accaggcctg ggcgaattcc ctcctcctcc
      961 gccaccgcca ccaccgccgc cgccgccacc accacctcct ccgcctcctc caccaccacc
     1021 gccaccgcct ccaccacctg gcggccgcat ggacaaggat tgtgaaatga aacgcaccac
     1081 actggacagc cctttgggga agctggagct gtctggttgt gagcagggtc tgcacaaaat
     1141 aatttttctg ggcaagggga cgtctgcagc tgatgccgtg gaggtcccag cccccgctgc
     1201 ggttctcgga ggtccggagc ccctgataca ggctacagcc tggctgaatg cctatttcca
     1261 ccagcccgag gctatcgaag agttccccgt gccggctctt caccatcccg ttttccagca
     1321 agagtcgttc accagacagg tgttatggaa gctgctgaag gttgtgaaat tcggagaagt
     1381 gatttctgag tcacacttag cagccctggt aggcaacccc gcagccacgg cagcagtgaa
     1441 tacggcactg gatggcaatc ctgtccctat cctgatcccg tgccacagag tggtccaggg
     1501 ggattcggat gtggggccgt accttggtgg actggccgtg aaggaatggc ttctggccca
     1561 tgaaggccac cggttgggga agccaggctt gggactcgag atgatcagtc tgattgcggc
     1621 gttagcggta gatcgcgtta tcggcatgga aaacgccatg ccgtggaacc tgcctgccga
     1681 tctcgcctgg tttaaacgca acaccttaaa taaacccgtg attatgggcc tccaaacctg
     1741 ggaatcaatc ggtcgtccgt tgccaggacg caaaaatatt atcctcagca gtcaaccggg
     1801 tacggacgat cgcgtaacgt gggtgaagtc ggtggatgaa gccatcgcgg cgtgtggtga
     1861 cgtaccagaa atcatggtga ttggcggcgg tcgcgtttat gaacagttct tgccaaaagc
     1921 gcaaaaactg tatctgacgc atatcgacgc agaagtggaa ggcgacaccc atttcccgga
     1981 ttacgagccg gatgactggg aatcggtatt cagcgaattc cacgatgctg atgcgcagaa
     2041 ctctcacagc tattgctttg agattctgga gcggcgggga gctccgggct tctcctcaat
     2101 ttccgctcat caccaccatc atcaccatca ccaccactaa ctcgagagct tggctgtttt
     2161 ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg
     2221 ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     2281 tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg
     2341 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     2401 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     2461 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     2521 tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt
     2581 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2641 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2701 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2761 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2821 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2881 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     2941 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3001 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3061 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3121 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3181 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3241 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3301 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3361 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3421 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3481 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3541 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3601 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3661 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3721 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3781 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3841 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3901 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3961 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4021 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4081 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4141 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4201 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4261 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4321 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4381 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4441 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     4501 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     4561 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     4621 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4681 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4741 cgaaacgcgc gaggcagcag atcaattcgc gcgcgaaggc gaagcggcat gcataatgtg
     4801 cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccgtc aagccgtcaa
     4861 ttgtctgatt cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac
     4921 aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat
     4981 agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc
     5041 tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc
     5101 ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga
     5161 cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc
     5221 gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta
     5281 acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg
     5341 cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa
     5401 agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg
     5461 gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat
     5521 gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct
     5581 gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca
     5641 gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca
     5701 ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac
     5761 ttttcata
//
