LOCUS       Exported                5966 bp ds-DNA     circular SYN 02-JUL-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5966)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 2, 2018 from SnapGene Viewer 4.1.9
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5966
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    173..313
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(499..1087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1194..2009)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        2123..2167
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    2143..2159
                     /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)."
     protein_bind    2175..2191
                     /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)."
     RBS             2210..2221
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2228..2902
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             3662..3679
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3746..3840
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3884..4543
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      4608..4694
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4787..5869)
                     /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"
     promoter        complement(5870..5947)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
ORIGIN
        1 ggtcctggcc acgggtgcgc atgatctaga gctgcctcgc gcgtttcggt gatgacggtg
       61 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
      121 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
      181 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
      241 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
      301 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      361 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      421 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      481 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      541 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      601 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      661 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      721 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      781 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      841 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      901 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
      961 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1021 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1081 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1141 acgttaaggg attttggtca tggcgttaag ggattttggt catgaattaa ttcttagaaa
     1201 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     1261 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     1321 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     1381 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     1441 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     1501 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     1561 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     1621 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     1681 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     1741 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     1801 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     1861 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     1921 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag
     1981 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     2041 gacagtttta ttgttcatga ccatgacatt aacctataaa aataggcgta tcacgaggcc
     2101 ctttcgtctt cacctcgaga aatcataaaa aatttatttg ctttgtgagc ggataacaat
     2161 tataatagat tcaattgtga gcggataaca atttcacaca gaattcatta aagaggagaa
     2221 attaactatg gcttcctccg aagacgttat caaagagttc atgcgtttca aagttcgtat
     2281 ggaaggttcc gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta
     2341 cgaaggtacc cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg
     2401 ggacatcctg tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga
     2461 catcccggac tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa
     2521 cttcgaagac ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt
     2581 catctacaaa gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa
     2641 aaaaaccatg ggttgggaag cttccaccga acgtatgtac ccggaagacg gtgctctgaa
     2701 aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa
     2761 aaccacctac atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa
     2821 actggacatc acctcccaca acgaagacta caccatcgtt gaacagtacg aacgtgctga
     2881 aggtcgtcac tccaccggtg ctatgggatc cgctggctcc gctgctggtt ctggcgaata
     2941 cgcatccaag ggcgaggagc tctttactgg cgtagtacca attctcgtag agctcgatgg
     3001 cgatgtaaat ggccataagt tttccgtacg cggcgagggc gagggcgatg caactaacgg
     3061 caagctcact ctcaagttta tttgtactac tggcaagctc ccagtaccat ggccaactct
     3121 cgtaactact ctgacctatg gcgtacaatg tttttcccgc tatccagatc acatgaagca
     3181 acatgatttt tttaagtccg caatgccaga gggctatgta caagagcgca ctattagctt
     3241 taaggatgat ggcacctata agactcgcgc agaggtaaag tttgagggcg atactctcgt
     3301 aaatcgcatt gagctcaagg gcattgattt taaggaggat ggcaatattc tcggccataa
     3361 gctggagtat aatttcaatt cccataatgt atatattacc gcagataagc aaaagaatgg
     3421 cattaaggcg aattttaaga ttcgccataa tgtggaggat ggctccgtac aactcgcaga
     3481 tcattatcaa caaaatactc caattggcga tggcccagta ctcctcccag ataatcatta
     3541 tctctccact caatccgtgc tctccaaaga tccaaatgag aagcgcgatc acatggtact
     3601 cctggagttt gtaactgcag caggcattac tcatggcatg gatgagctct ataagctcga
     3661 gcaccaccac caccaccact aaggatccgc atgcgagctc ggtaccccgg gtcgacctgc
     3721 agccaagctt aattagctga gcttggactc ctgttgatag atccagtaat gacctcagaa
     3781 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat
     3841 ccaagctagc ttggcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac
     3901 tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca
     3961 gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa
     4021 gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct
     4081 gatgaatgct catccggaat ttcgtatggc aatgaaagac ggtgagctgg tgatatggga
     4141 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg
     4201 gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg
     4261 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc
     4321 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt
     4381 cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
     4441 gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa
     4501 tgaattacaa cagtactgcg atgagtggca gggcggggcg taattttttt aaggcagtta
     4561 ttggtgccct taaacgcctg gggtaatgac tctctagctt gaggcatcaa ataaaacgaa
     4621 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     4681 tgagtaggac aaatccgccc tctagattac gtgcagtcga tgataagctg tcaaacatga
     4741 gaattgtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     4801 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4861 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     4921 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     4981 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     5041 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
     5101 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     5161 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     5221 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     5281 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     5341 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     5401 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     5461 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     5521 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     5581 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     5641 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     5701 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     5761 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     5821 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     5881 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg caccattcga
     5941 tggtgtcgga atttcgggca gcgttg
//
