LOCUS       pSD117A                 6565 bp ds-DNA     circular SYN 13-MAR-2024
DEFINITION  pET AviTag-6xHis-MBP-TEV-(GGGGS)2-SD40 KanR.
ACCESSION
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid.
REFERENCE   1  (bases 1 to 11577)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Oct 30, 2018 from SnapGene 4.2.5 http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6565
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     rep_origin      12..467
     primer_bind     complement(99..118)
                     /note="F1 origin, reverse primer"
                     /standard_name="F1ori-R"
     primer_bind     309..330
                     /note="F1 origin, forward primer"
                     /standard_name="F1ori-F"
     CDS             complement(559..1374)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="confers resistance to kanamycin in bacteria or G418
                      (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
                     /standard_name="KanR"
     misc_feature    complement(956..980)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="JM116"
     misc_feature    964..980
                     /standard_name="USER"
     primer_bind     964..986
                     /standard_name="JM115"
     primer_bind     1282..1301
                     /note="Kanamycin resistance gene, reverse primer"
                     /standard_name="Kan-R"
     primer_bind     1366..1385
                     /note="pENTR vectors, reverse primer"
                     /standard_name="pENTR-R"
     rep_origin      1496..2084
     primer_bind     1985..2004
                     /note="pBR322 origin, forward primer"
                     /standard_name="pBR322ori-F"
     primer_bind     2238..2255
                     /note="L4440 vector, forward primer"
                     /standard_name="L4440"
     misc_feature    2270..2409
                     /note="basis of mobility region from pBR322"
                     /standard_name="bom"
     primer_bind     complement(2335..2354)
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
                     /standard_name="pRS-marker"
     primer_bind     2495..2517
                     /note="pGEX vectors, reverse primer"
                     /standard_name="pGEX 3'"
     CDS             complement(2511..2702)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low
                     copy number"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH
                     ADELYRSCLARFGDDGENL"
                     /standard_name="rop"
     CDS             complement(3511..4593)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
                     LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
                     GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
                     DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
                     EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
                     QTASPRALADSLMQLARQVSRLESGQ"
                     /standard_name="lacI"
     primer_bind     4555..4574
                     /note="LacI, reverse primer"
                     /standard_name="LacI-R"
     regulatory      complement(4594..4671)
                     /note="Geneious type: promoter"
                     /gene="lacI"
     primer_bind     4877..4896
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
                     /standard_name="pBRrevBam"
     regulatory      4980..4998
                     /note="Geneious type: promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /standard_name="T7 promoter"
     primer_bind     4980..4999
                     /note="T7 promoter, forward primer"
                     /standard_name="T7"
     protein_bind    4999..5023
                     /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)."
                     /standard_name="lac operator"
     primer_bind     complement(5030..5089)
                     /standard_name="SD417"
     primer_bind     complement(5030..5070)
                     /standard_name="SD097"
     misc_feature    5038..5060
                     /note="Geneious type: RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    5065..>5067
                     /note="Geneious type: restriction"
                     /standard_name="NdeI"
     CDS             5068..6429
                     /standard_name="AviTag-6xHis-MBP-TEV-Super"
     gene            5071..5115
                     /standard_name="AviTag"
     source          <5071..>5115
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     5090..5140
                     /standard_name="SD416"
     gene            5125..5142
                     /standard_name="6xHis"
     primer_bind     5130..5181
                     /standard_name="SD373"
     gene            5152..6261
                     /standard_name="MBP"
     source          <5152..>6261
     source          join(<5152..>6261,<5152..>6261)
     primer_bind     5229..5250
                     /standard_name="TW0202"
     misc_difference 5245..5250
                     /standard_name
     gene            6262..6288
                     /standard_name="GS-TEV"
     primer_bind     complement(6262..6303)
                     /standard_name="SD379"
     source          <6289..>6291
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     6304..6342
                     /standard_name="SD378"
     gene            6319..6426
                     /standard_name="Super"
     misc_feature    6430..6456
                     /note="Geneious type: overhang"
                     /standard_name="Gibson"
     primer_bind     6430..6457
                     /standard_name="SD139"
     primer_bind     complement(6479..6497)
                     /note="T7 terminator, reverse primer"
                     /standard_name="T7 Term"
     regulatory      6493..6540
                     /note="Geneious type: terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /standard_name="T7 terminator"
ORIGIN      
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
      481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      541 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      601 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
      661 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
      721 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
      781 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
      841 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
      901 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
      961 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     1021 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     1081 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     1141 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     1201 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     1261 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1321 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc
     1381 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac
     1441 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1501 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1561 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1621 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     1681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1981 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2041 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2101 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2161 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     2221 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     2281 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     2341 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     2401 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2461 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     2521 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     2581 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     2641 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     2701 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     2761 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     2821 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     2881 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     2941 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     3001 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     3061 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     3121 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     3181 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     3241 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     3301 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     3361 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     3421 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     3481 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3541 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3601 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     3661 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     3721 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     3781 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     3841 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     3901 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     3961 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     4021 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     4081 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     4141 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     4201 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     4261 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     4321 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     4381 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     4441 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     4501 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     4561 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     4621 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     4681 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     4741 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     4801 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     4861 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     4921 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     4981 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
     5041 gtttaacttt aagaaggaga tatacatatg ggcttaaacg acatttttga ggctcaaaag
     5101 atcgaatggc acgagggcag ctctcatcat catcatcacc acggttccag taagatcgaa
     5161 gaaggcaagt tagtgatttg gatcaacggc gataaaggct ataacggttt agcggaagta
     5221 ggcaagaagt ttgagaagga taccggaatc aaagtgacgg tcgaacaccc ggataagtta
     5281 gaggagaagt tccctcaagt agcggcaacg ggcgacggtc ctgacatcat cttttgggca
     5341 cacgaccgct tcggtggtta cgctcagtca ggtttattag cggagattac tccagacaaa
     5401 gcgttccagg acaagttata tccatttacg tgggacgcag tgcgttataa cggcaagtta
     5461 atcgcatatc ccattgcggt agaagccctg agcctgattt acaacaagga cctgttaccc
     5521 aaccctccga agacgtggga ggagattccg gctttagata aggaactgaa agcgaaaggc
     5581 aagtctgccc tgatgtttaa cctgcaagaa ccctacttca cctggcctct gatcgcggcg
     5641 gatggcggtt atgcgtttaa gtacgaaaac ggcaaatacg atattaagga tgtaggcgtc
     5701 gataacgccg gtgccaaagc cggtctgacc tttttagttg atctgattaa gaataagcac
     5761 atgaacgcag acaccgacta cagcattgcg gaagctgcgt ttaataaagg tgagaccgcc
     5821 atgacgatta acggaccttg ggcgtggtcg aacattgata ccagtaaagt caattacggc
     5881 gttacagtcc tgccgacctt caaaggtcag ccgtcaaaac cgttcgtagg tgtcttatcc
     5941 gccggtatca acgcggcgtc cccaaataaa gagttggcta aagagttcct ggaaaactat
     6001 ctgctgacag acgaaggact ggaggctgtg aacaaggaca agccactggg cgctgttgcg
     6061 ctgaaaagtt atgaggaaga actggcgaaa gatccccgca tcgcggcgac gatggaaaac
     6121 gcccaaaaag gcgaaatcat gccgaacatt ccgcagatgt cagctttttg gtatgccgta
     6181 cgcacggctg ttattaacgc cgcgtcgggc cgccaaaccg ttgatgaggc actgaaggac
     6241 gcgcagactc gtatcaccaa gggtagcgaa aatctttatt ttcaaggctc gggtggaggg
     6301 ggctcgggag gtggcggtct gttgctgttc tgccctattt gcgggtttac atgtcgccag
     6361 aagggcaact tacttcgcca tattaacctg cacacagggg aaaagttatt taagtaccac
     6421 ctgtattgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     6481 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     6541 ctgaaaggag gaactatatc cggat
//
