LOCUS       Exported                4413 bp ds-DNA     circular SYN 06-OCT-2014
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCDF-Sfp
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4413)
  AUTHORS   Nic Harmer
  TITLE     Direct Submission
  JOURNAL   Exported 23 Sep 2016 from SnapGene 2.6.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4413
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    3..27
                     /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)."
     CDS             71..745
                     /codon_start=1
                     /note="Sfp-Transferase"
                     /translation="MKIYGIYMDRPLSQEENERFMTFISPEKREKCRRFYHKEDAHRTL
                     LGDVLVRSVISRQYQLDKSDIRFSTQEYGKPCIPDLPDAHFNISHSGRWVIGAFDSQPI
                     GIDIEKTKPISLEIAKRFFSKTEYSDLLAKDKDEQTDYFYHLWSMKESFIKQEGKGLSL
                     PLDSFSVRLHQDGQVSIELPDSHSPCYIKTYEVDPGYKMAVCAAHPDFPEDITMVSYEE
                     LL"
     promoter        846..864
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    865..889
                     /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)."
     misc_feature    930..1065
                     /note="MCS2"
     CDS             998..1042
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /note="S-Tag"
                     /translation="KETAAAKFERQHMDS"
     terminator      1094..1141
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1312..2103)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /note="SmR"
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2104..2195)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      complement(2243..2981)
                     /direction=LEFT
                     /note="CloDF13 ori"
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(3191..4273)
                     /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(4274..4351)
                     /gene="lacI"
                     /note="lacI promoter"
     promoter        4397..2
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgaagattt acggaattta tatggaccgc ccgctttcac aggaagaaaa
      121 tgaacggttc atgactttca tatcacctga aaaacgggag aaatgccgga gattttatca
      181 taaagaagat gctcaccgca ccctgctggg agatgtgctc gttcgctcag tcataagcag
      241 gcagtatcag ttggacaaat ccgatatccg ctttagcacg caggaatacg ggaagccgtg
      301 catccctgat cttcccgacg ctcatttcaa catttctcac tccggccgct gggtcattgg
      361 tgcgtttgat tcacagccga tcggcataga tatcgaaaaa acgaaaccga tcagccttga
      421 gatcgccaag cgcttctttt caaaaacaga gtacagcgac cttttagcaa aagacaagga
      481 cgagcagaca gactattttt atcatctatg gtcaatgaaa gaaagcttta tcaaacagga
      541 aggcaaaggc ttatcgcttc cgcttgattc cttttcagtg cgcctgcatc aggacggaca
      601 agtatccatt gagcttccgg acagccattc cccatgctat atcaaaacgt atgaggtcga
      661 tcccggctac aaaatggctg tatgcgccgc acaccctgat ttccccgagg atatcacaat
      721 ggtctcgtac gaagagcttt tataaattcg agctcggcgc gcctgcaggt cgacaagctt
      781 gcggccgcat aatgcttaag tcgaacagaa agtaatcgta ttgtacacgg ccgcataatc
      841 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc catcttagta
      901 tattagttaa gtataagaag gagatataca tatggcagat ctcaattgga tatcggccgg
      961 ccacgcgatc gctgacgtcg gtaccctcga gtctggtaaa gaaaccgctg ctgcgaaatt
     1021 tgaacgccag cacatggact cgtctactag cgcagcttaa ttaacctagg ctgctgccac
     1081 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1141 gctgaaacct caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc
     1201 ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgaccgg
     1261 gtcatcgtgg ccggatcttg cggcccctcg gcttgaacga attgttagac attatttgcc
     1321 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg
     1381 cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg
     1441 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
     1501 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
     1561 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
     1621 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
     1681 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
     1741 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
     1801 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
     1861 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
     1921 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
     1981 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
     2041 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct
     2101 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     2161 atacatattt gaatgtattt agaaaaataa acaaatagct agctcactcg gtcgctacgc
     2221 tccgggcgtg agactgcggc gggcgctgcg gacacataca aagttaccca cagattccgt
     2281 ggataagcag gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc
     2341 cataggctcc gccctcctgc cagagttcac ataaacagac gcttttccgg tgcatctgtg
     2401 ggagccgtga ggctcaacca tgaatctgac agtacgggcg aaacccgaca ggacttaaag
     2461 atccccaccg tttccggcgg gtcgctccct cttgcgctct cctgttccga ccctgccgtt
     2521 taccggatac ctgttccgcc tttctccctt acgggaagtg tggcgctttc tcatagctca
     2581 cacactggta tctcggctcg gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac
     2641 tccccgttca gcccgactgc tgcgccttat ccggtaactg ttcacttgag tccaacccgg
     2701 aaaagcacgg taaaacgcca ctggcagcag ccattggtaa ctgggagttc gcagaggatt
     2761 tgtttagcta aacacgcggt tgctcttgaa gtgtgcgcca aagtccggct acactggaag
     2821 gacagatttg gttgctgtgc tctgcgaaag ccagttacca cggttaagca gttccccaac
     2881 tgacttaacc ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga
     2941 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct
     3001 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
     3061 agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     3121 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     3181 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3241 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     3301 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     3361 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     3421 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
     3481 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     3541 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     3601 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     3661 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     3721 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     3781 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     3841 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     3901 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     3961 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     4021 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     4081 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     4141 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     4201 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     4261 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     4321 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     4381 tcctgcatta ggaaattaat acgactcact ata
//
