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    167..307
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(493..1081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1188..2003)
                     /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        2117..2161
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    2137..2153
                     /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    2169..2185
                     /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             2204..2215
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2222..2896
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             3656..3673
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3740..3834
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3878..4537
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      4602..4688
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4781..5863)
                     /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(5864..5941)
                     /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 ggccacgggt gcgcatgatc tagagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
       61 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
      121 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
      181 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
      241 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
      301 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
      361 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
      421 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
      481 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
      541 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
      601 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
      661 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
      721 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
      781 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
      841 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
      901 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
      961 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1021 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1081 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1141 agggattttg gtcatggcgt taagggattt tggtcatgaa ttaattctta gaaaaactca
     1201 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     1261 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     1321 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     1381 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     1441 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     1501 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     1561 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     1621 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     1681 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     1741 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     1801 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     1861 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     1921 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac
     1981 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     2041 tttattgttc atgaccatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     2101 tcttcacctc gagaaatcat aaaaaattta tttgctttgt gagcggataa caattataat
     2161 agattcaatt gtgagcggat aacaatttca cacagaattc attaaagagg agaaattaac
     2221 tatggcttcc tccgaagacg ttatcaaaga gttcatgcgt ttcaaagttc gtatggaagg
     2281 ttccgttaac ggtcacgagt tcgaaatcga aggtgaaggt gaaggtcgtc cgtacgaagg
     2341 tacccagacc gctaaactga aagttaccaa aggtggtccg ctgccgttcg cttgggacat
     2401 cctgtccccg cagttccagt acggttccaa agcttacgtt aaacacccgg ctgacatccc
     2461 ggactacctg aaactgtcct tcccggaagg tttcaaatgg gaacgtgtta tgaacttcga
     2521 agacggtggt gttgttaccg ttacccagga ctcctccctg caagacggtg agttcatcta
     2581 caaagttaaa ctgcgtggta ccaacttccc gtccgacggt ccggttatgc agaaaaaaac
     2641 catgggttgg gaagcttcca ccgaacgtat gtacccggaa gacggtgctc tgaaaggtga
     2701 aatcaaaatg cgtctgaaac tgaaagacgg tggtcactac gacgctgaag ttaaaaccac
     2761 ctacatggct aaaaaaccgg ttcagctgcc gggtgcttac aaaaccgaca tcaaactgga
     2821 catcacctcc cacaacgaag actacaccat cgttgaacag tacgaacgtg ctgaaggtcg
     2881 tcactccacc ggtgctatgg gatccgctgg ctccgctgct ggttctggcg aataggcatc
     2941 caagggcgag gagctcttta ctggcgtagt accaattctc gtagagctcg atggcgatgt
     3001 aaatggccat aagttttccg tacgcggcga gggcgagggc gatgcaacta acggcaagct
     3061 cactctcaag tttatttgta ctactggcaa gctcccagta ccatggccaa ctctcgtaac
     3121 tactctgacc tatggcgtac aatgtttttc ccgctatcca gatcacatga agcaacatga
     3181 tttttttaag tccgcaatgc cagagggcta tgtacaagag cgcactatta gctttaagga
     3241 tgatggcacc tataagactc gcgcagaggt aaagtttgag ggcgatactc tcgtaaatcg
     3301 cattgagctc aagggcattg attttaagga ggatggcaat attctcggcc ataagctgga
     3361 gtataatttc aattcccata atgtatatat taccgcagat aagcaaaaga atggcattaa
     3421 ggcgaatttt aagattcgcc ataatgtgga ggatggctcc gtacaactcg cagatcatta
     3481 tcaacaaaat actccaattg gcgatggccc agtactcctc ccagataatc attatctctc
     3541 cactcaatcc gtgctctcca aagatccaaa tgagaagcgc gatcacatgg tactcctgga
     3601 gtttgtaact gcagcaggca ttactcatgg catggatgag ctctataagc tcgagcacca
     3661 ccaccaccac cactaaggat ccgcatgcga gctcggtacc ccgggtcgac ctgcagccaa
     3721 gcttaattag ctgagcttgg actcctgttg atagatccag taatgacctc agaactccat
     3781 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc
     3841 tagcttggcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
     3901 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     3961 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     4021 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     4081 tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     4141 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     4201 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     4261 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     4321 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     4381 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     4441 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     4501 acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg
     4561 cccttaaacg cctggggtaa tgactctcta gcttgaggca tcaaataaaa cgaaaggctc
     4621 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     4681 ggacaaatcc gccctctaga ttacgtgcag tcgatgataa gctgtcaaac atgagaattg
     4741 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4801 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4861 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     4921 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     4981 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     5041 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
     5101 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     5161 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     5221 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     5281 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     5341 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     5401 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     5461 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     5521 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     5581 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     5641 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     5701 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     5761 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     5821 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     5881 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcaccat tcgatggtgt
     5941 cggaatttcg ggcagcgttg ggtcct
//
