LOCUS       Exported                5918 bp ds-DNA     circular SYN 30-NOV-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pGrpE-Wasabi
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5918)
  AUTHORS   Dan Piraner
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Nov 30, 2016 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5918
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        47..168
                     /note="GrpE Promoter"
                     /note="From NC_000913.3"
     CDS             169..171
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     CDS             181..198
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             211..921
                     /codon_start=1
                     /product="green variant of teal fluorescent protein mTFP1 
                     (Ai et al., 2008)"
                     /note="mWasabi"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFSYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKETTGWDASTERMYVRDGVLK
                     GDVKMKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYEIA
                     VARNSTDGMDELYKG"
     terminator      1023..1070
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1107..1562
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1677..2537)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2538..2630)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      2711..3299
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3729..3920)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4492..5574)
                     /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"
     promoter        complement(5575..5652)
                     /gene="lacI"
                     /note="lacI promoter"
ORIGIN
        1 atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg cgaaattttt ccgaggtcct
       61 tgttgcgaag attgatgaca atgtgagtgc ttcccttgaa accctgaaac tgatccccat
      121 aataagcgaa gttagcgaga tgaatgcgaa aaaaacgcgg agaaattcat gggcagcagc
      181 catcaccatc atcaccacag ccaggatcca atggtgagca agggcgagga gaccacaatg
      241 ggcgtaatca agcccgacat gaagatcaag ctgaagatgg agggcaacgt gaatggccac
      301 gccttcgtga tcgagggcga gggcgagggc aagccctacg acggcaccaa caccatcaac
      361 ctggaggtga aggagggagc ccccctgccc ttctcctacg acattctgac caccgcgttc
      421 agttacggca acagggcctt caccaagtac cccgacgaca tccccaacta cttcaagcag
      481 tccttccccg agggctactc ttgggagcgc accatgacct tcgaggacaa gggcatcgtg
      541 aaggtgaagt ccgacatctc catggaggag gactccttca tctacgagat acacctcaag
      601 ggcgagaact tcccccccaa cggccccgtg atgcagaagg agaccaccgg ctgggacgcc
      661 tccaccgaga ggatgtacgt gcgcgacggc gtgctgaagg gcgacgtcaa gatgaagctg
      721 ctgctggagg gcggcggcca ccaccgcgtt gacttcaaga ccatctacag ggccaagaag
      781 gcggtgaagc tgcccgacta tcactttgtg gaccaccgca tcgagatcct gaaccacgac
      841 aaggactaca acaaggtgac cgtttacgag atcgccgtgg cccgcaactc caccgacggc
      901 atggacgagc tgtacaaggg ctgagtaaag aaaccgctgc tgcgaaattt gaacgccagc
      961 acatggactc gtctactagc gcagcttaat taacctaggc tgctgccacc gctgagcaat
     1021 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     1081 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     1141 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1201 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1261 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1321 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1381 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1441 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1501 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     1561 ttctggcggc acgatggcat gagattatca aaaaggatct tcacctagat ccttttaaat
     1621 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     1681 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1741 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1801 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     1861 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1921 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1981 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2041 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2101 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2161 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     2221 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     2281 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     2341 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     2401 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     2461 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     2521 aaatgttgaa tactcatact cttccttttt caatcatgat tgaagcattt atcagggtta
     2581 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggtcatga
     2641 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2701 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2761 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2821 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     2881 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2941 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3001 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3061 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3121 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3181 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3241 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3301 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3361 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3421 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3481 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatatg
     3541 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     3601 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     3661 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3721 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     3781 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     3841 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     3901 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     3961 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     4021 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     4081 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     4141 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     4201 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     4261 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     4321 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcta
     4381 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     4441 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     4501 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4561 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     4621 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     4681 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     4741 gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc
     4801 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     4861 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     4921 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     4981 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     5041 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     5101 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     5161 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     5221 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     5281 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     5341 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     5401 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     5461 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     5521 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     5581 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     5641 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     5701 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     5761 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     5821 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     5881 cagcaaccgc acctgtggcg ccggtgatgc cggccacg
//
