LOCUS       Exported                7443 bp ds-DNA     circular SYN 31-MAY-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pXM-IDS2-ximB
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7443)
  AUTHORS   Hebb
  TITLE     Direct Submission
  JOURNAL   Exported Dec 10, 2018 from SnapGene Viewer 4.2.9
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7443
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     terminator      26..73
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     157..183
                     /label=ximB R
     misc_feature    158
                     /label=MCS
                     /note="multiple cloning site"
     CDS             complement(164..1105)
                     /codon_start=1
                     /label=XimB
                     /translation="VIDISAQPSQQSTRYRFVVPSIAASVVPSAPKAVLPYLQLARMHA
                     PIGSWLYLLPGLWGIALASAGLPDWRQVLLFTVGGVLVRGFGCVVNDLADRKFDARVAR
                     TVGRPLVAGTVTVTGALVFAVVQAVAGLLVLAAASVPAAVFVAASYPLVVAYPFMKRIT
                     YWPQAWLGMVFGCYILAGWLAVAGRIETPALLLFVAGVFWTLGYDTIYAHQDKADDVQV
                     GVKSLALRLGRATRPWVAGFYGATVVGIVAAGAVADLHWTFYALFLPGVAHLLWQVVTV
                     DIDSPADCREKFMANRFFGWLVLVAIVAGRVF"
     primer_bind     complement(1086..1111)
                     /label=ximB F
     CDS             complement(1108)
                     /codon_start=1
                     /product="epitope tag from a T7 major capsid protein"
                     /label=T7 tag
                     /translation=""
     RBS             1117..1122
                     /note="ribosome binding site"
     protein_bind    1153..1177
                     /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)."
     promoter        complement(1178..1196)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1221..1268
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1335..1352)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    1353..1368
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     1367..1394
                     /label=GPPS R
     CDS             complement(1374..2330)
                     /codon_start=1
                     /label=IDS2
                     /translation="MTRSSKALVQLADLSEQVKNVVEFDFDKYMHSKAIAVNEALDKVI
                     PPRYPQKIYESMRYSLLAGGKRVRPILCIAACELMGGTEELAMPTACAIEMIHTMSLIH
                     DDLPYIDNDDLRRGKPTNHKVFGEDTAIIAGDALLSLAFEHVAVSTSRTLGTDIILRLL
                     SEIGRATGSEGVMGGQVVDIESEGDPSIDLETLEWVHIHKTAVLLECSVVCGAIMGGAS
                     EDDIERARRYARCVGLLFQVVDDILDVSQSSEELGKTAGKDLISDKATYPKLMGLEKAK
                     EFADELLNRGKQELSCFDPTKAAPLFALADYIASRQN"
     primer_bind     complement(2312..2337)
                     /label=GPPS F
     misc_feature    2333..2337
                     /label=MCS
                     /note="multiple cloning site"
     CDS             complement(2340..2372)
                     /codon_start=1
                     /product="epitope tag from a T7 major capsid protein"
                     /label=T7 tag
                     /translation="MASMTGGQQMG"
     RBS             2381..2386
                     /note="ribosome binding site"
     protein_bind    2417..2441
                     /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)."
     promoter        complement(2442..2460)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2769..2846
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2847..3929
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             4738..4929
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(5359..5947)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             6069..6884
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(6977..7432)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggaact agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtcaaaag actctccccg
      181 caacgatggc gacgagcacg agccacccga agaaacggtt tgccatgaac ttttcccggc
      241 agtcggcagg gctgtcgatg tccaccgtca ccacctgcca gagcaggtgt gccacgcccg
      301 gcagaaacag cgcgtagaac gtccagtgca gatcggcgac ggctccggca gccacgatcc
      361 cgaccacggt cgccccgtaa aacccggcga cccagggccg ggtcgccctg ccgaggcgca
      421 gcgcgaggga cttcaccccc acctgcacgt cgtccgcctt gtcctggtgg gcgtagatcg
      481 tgtcgtaccc cagcgtccag aagacaccgg cgacgaagag caggagcgcg ggagtctcga
      541 tcctcccggc caccgcgagc caacccgcga ggatgtagca gccgaagacc atccccagcc
      601 aggcctgcgg ccagtaggtg atccgcttca tgaacgggta cgcgaccacg agcggatagg
      661 aggcggccac gaacaccgcc gccggaacgc tggccgcagc cagcacgagc agccccgcca
      721 ccgcctgcac cacggcgaag acaagcgctc ccgtgaccgt gaccgtaccg gccacgagcg
      781 gcctcccgac ggtacgtgcc acccgggcgt cgaacttgcg gtcggcaagg tcgttgacga
      841 cgcagccgaa gccgcgcacc agcaccccac cgaccgtgaa cagcagcacc tggcgccagt
      901 cgggcagccc cgccgaggcg agcgcgatcc cccacaggcc ggggaggagg tacagccaac
      961 tgcctatggg cgcgtgcata cgcgccagtt gtagataggg gaggacggcc ttcggcgcac
     1021 tcggcacgac cgacgccgcg atgctcggca cgacgaaccg atatcgcgtg ctctgctgcg
     1081 agggttgagc ggaaatatcg atcaccatat gtatatctcc ttcttaaagt taaacaaaat
     1141 tatttgtaga ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt
     1201 tttctagttc ctcctttcag caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     1261 tatgctagtt attgctcagc ggtggcagca gccaactcag cttcctttcg ggctttgtta
     1321 gcagccggat ctcagtggtg gtggtggtgg tgctcgagtg cggccgcaag cttttaattc
     1381 tgacgagatg caatgtagtc tgctaacgca aatagaggtg ctgctttggt aggatcaaaa
     1441 caactaagtt cctgttttcc acggttcagt aattcatcgg caaattcctt cgctttttcc
     1501 aaacccatga gtttgggata ggtggctttg tcagaaatca aatctttccc agcagtcttt
     1561 ccgagttctt ccgaggactg gcttacatcc aaaatatcat cgacaacttg gaaaagcaat
     1621 cctacacagc gagcgtaccg tctagcacgc tcgatgtcgt cctcgctggc accccccata
     1681 attgcgccac acacgacact gcattccaac aacacagccg ttttatgaat atggacccat
     1741 tccagcgttt ctaagtctat actgggatca ccttcgcttt caatatccac cacttgacca
     1801 cccatcacgc cctcacttcc tgtggcgcgt ccaatttcgg atagcaaccg taaaataatg
     1861 tcagtaccta gggtacgact ggtgctcacg gctacatgtt caaaggccaa tgacaataat
     1921 gcatcgccag caatgatcgc cgtgtcttca ccaaaaactt tgtggttggt aggcttaccg
     1981 cgacgcaaat catcgttatc aatatagggc aaatcgtcat gaatcaaact catagtgtga
     2041 atcatctcga tggcacaagc cgtaggcatg gcaagttcct cagtcccccc cattagctca
     2101 caggccgcaa tacataaaat tggtcgaacc ctcttcccgc cggctaggag ggaatagcgc
     2161 atactttcat agattttttg aggatagcgg gggggaataa ctttatctaa ggcctcatta
     2221 accgcaatgg ccttggagtg catatacttg tcaaaatcaa attccaccac gttttttact
     2281 tgttcggata gatcagctag ttggaccaag gccttactgc tgcgcgtcat cggatcccga
     2341 cccatttgct gtccaccagt catgctagcc atatgtatat ctccttctta aagttaaaca
     2401 aaattatttg tagaggggaa ttgttatccg ctcacaattc ccctatagtg agtcgtatta
     2461 attttttcta gacgagatct cgatcctcta cgccggacgc atcgtggccg gcatcaccgg
     2521 cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc
     2581 tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc
     2641 cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa
     2701 cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg
     2761 agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg
     2821 aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt
     2881 atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg
     2941 cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg
     3001 tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg
     3061 ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg
     3121 ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc
     3181 acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg
     3241 atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg
     3301 accagacacc catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg
     3361 agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg
     3421 tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc
     3481 cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa
     3541 tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg
     3601 gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg
     3661 gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg
     3721 attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg
     3781 cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc
     3841 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3901 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact
     3961 cattaggcac cgggatctcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     4021 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     4081 ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg
     4141 agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa
     4201 gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc
     4261 atggcggccc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg
     4321 cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac
     4381 tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt
     4441 tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg gatctgcatc
     4501 gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat
     4561 tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca
     4621 caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct
     4681 cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg
     4741 accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg
     4801 cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt
     4861 cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa
     4921 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     4981 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     5041 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
     5101 ttgtactgag agtgcaccat atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     5161 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5221 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5281 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5341 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5401 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5461 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5521 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5581 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5641 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5701 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5761 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     5821 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5881 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5941 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6001 acgttaaggg attttggtca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     6061 ggggtgttat gagccatatt caacgggaaa cgtcttgctc taggccgcga ttaaattcca
     6121 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     6181 cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     6241 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     6301 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     6361 ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     6421 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     6481 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     6541 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     6601 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     6661 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     6721 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     6781 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     6841 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaagaatta attcatgagc
     6901 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6961 cgaaaagtgc cacctgaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     7021 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     7081 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     7141 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     7201 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     7261 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     7321 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     7381 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     7441 cca
//
