LOCUS       Exported                6019 bp ds-DNA     circular SYN 18-MAR-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    ALARMNMR-plasmid
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6019)
  AUTHORS   Jayme Dahlin
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Mar 28, 2017 from SnapGene Viewer 3.3.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6019
                     /organism="recombinant plasmid"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     terminator      26..73
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(137..949)
                     /codon_start=1
                     /note="La antigen"
                     /translation="MGSSHHHHHHSSGLVPRGSHMASMTGGQQMGRDPVTDEYKNDVKN
                     RSVYIKGFPTDATLDDIKEWLEDKGQVLNIQMRRTLHKAFKGSIFVVFDSIESAKKFVE
                     TPGQKYKETDLLILFKDDYFAKKNEERKQNKVEAKLRAKQEQEAKQKLEEDAEMKSLEE
                     KIGCLLKFSGDLDDQTCREDLHILFSNHGEIKWIDFVRGAKEGIILFKEKAKEALGKAK
                     DANNGNLQLRNKEVTWEVLEGEVEKEALKKIIEDQQEAAALEHHHHHH"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     misc_feature    158..166
                     /note="MCS"
                     /note="multiple cloning site"
     misc_feature    850..854
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(857..889)
                     /codon_start=1
                     /product="epitope tag from a T7 major capsid protein"
                     /note="T7 tag"
                     /translation="MASMTGGQQMG"
     CDS             complement(893..910)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             complement(920..937)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             complement(947..949)
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     RBS             957..962
                     /note="ribosome binding site"
     protein_bind    993..1017
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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(1018..1036)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1345..1422
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             1423..2505
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="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             3314..3505
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(3935..4523)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4645..5460
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(5553..6008)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcttcttgtt
      181 ggtcttctat tattttcttc agtgcttctt tttccacctc tccttctagt acttcccaag
      241 tcacttcttt gttccttaat tgtaggttac cattatttgc atctttggct ttacccaatg
      301 cttccttggc tttttcttta aatagaatta tcccctcttt tgctcctctg acgaagtcta
      361 tccattttat ttcaccatga tttgagaaaa gtatgtgtaa atcttctcta caggtctgat
      421 catctaaatc acccgaaaat ttcagcaagc atccaatctt ttcttctaga gatttcattt
      481 cagcatcttc ttctaacttt tgttttgctt cttgctcctg tttagctctt aatttagctt
      541 ccactttatt ttgttttctt tcttcatttt ttttggcaaa gtaatcgtcc ttgaaaagta
      601 ttagcaggtc tgtttctttg tacttctggc caggggtctc tacaaatttc ttagcagatt
      661 caatgctatc aaacacaaca aaaattgatc ccttaaatgc tttatgcaat gttcttctca
      721 tctgaatatt tagtacttga cctttatctt ctaaccattc ttttatgtca tcaagagttg
      781 catcagttgg gaagccttta atataaacag atctgttttt tacatcattt ttatactcat
      841 cagtcaccgg atcccgaccc atttgctgtc caccagtcat gctagccata tggctgccgc
      901 gcggcaccag gccgctgctg tgatgatgat gatgatggct gctgcccatg gtatatctcc
      961 ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca caattcccct
     1021 atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc ggacgcatcg
     1081 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg
     1141 atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg
     1201 tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg
     1261 cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc
     1321 ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt tcgcggtatg
     1381 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
     1441 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     1501 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     1561 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     1621 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     1681 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     1741 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     1801 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     1861 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
     1921 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     1981 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     2041 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     2101 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     2161 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     2221 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta
     2281 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     2341 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
     2401 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2461 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2521 tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa
     2581 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     2641 cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga
     2701 ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt
     2761 gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca
     2821 ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag
     2881 gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga
     2941 gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt
     3001 cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat
     3061 gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt
     3121 aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc
     3181 cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat
     3241 cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta
     3301 cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag
     3361 aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga
     3421 catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt
     3481 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3541 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     3601 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     3661 gcggcatcag agcagattgt actgagagtg caccatatat gcggtgtgaa ataccgcaca
     3721 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
     3781 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3841 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3901 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3961 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4021 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4081 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4141 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4201 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4261 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4321 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     4381 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4441 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4501 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4561 agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact gtctgcttac
     4621 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc ttgctctagg
     4681 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     4741 gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg
     4801 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     4861 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     4921 gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt attagaagaa
     4981 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     5041 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     5101 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     5161 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     5221 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     5281 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     5341 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     5401 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     5461 gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5521 tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata ttttgttaaa
     5581 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5641 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5701 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5761 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5821 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5881 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5941 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6001 gggcgcgtcc cattcgcca
//
