LOCUS       Exported                7119 bp ds-DNA     circular SYN 29-NOV-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    p481-pCDest-AcrE2 BFP2 IRES
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7119)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Nov 29, 2016 from SnapGene 3.1.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7119
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        35..53
                     /note="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    106..357
                     /note="AcrE2"
     promoter        complement(376..393)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    398..532
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     enhancer        789..1168
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1169..1380
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          1541..1673
                     /note="chimeric intron"
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1718..1736
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1742..2452
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /note="mTagBFP2"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     misc_feature    2506..3079
                     /note="IRES"
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     promoter        complement(3128..3145)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    3163..3284
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(3415..3433)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3454..3470)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3478..3494
                     /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(3502..3532)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3547..3568
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3856..4444)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4615..5475)
                     /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(5476..5580)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      complement(5606..6061)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6135..7119
                     /note="CMV IE94 promoter"
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaagccg ccaccatgaa tacctatctc
      121 atcgaccccc gcaaaaacaa cgacaactcc ggcgagcgct tcacggttga cgctgtcgac
      181 attacagccg ccgcgaagag cgcagcccaa cagattcttg gcgaggaatt cgagggcctc
      241 gtataccgtg aaaccgggga gagtaacgga agtggcatgt tccaggccta ccaccacctg
      301 cacggcacta accgcacgga gacgaccgtt ggctatccgt ttcatgtaat ggaactctag
      361 ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
      421 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
      481 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
      541 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc
      601 ggccgcggcg ccaatgcatt gggcccggta ctcacatggc tcgacagatc ttcaatattg
      661 gccattagcc atattattca ttggttatat agcataaatc aatattggct attggccatt
      721 gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc
      781 gccatgttgg cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      841 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      901 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      961 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     1021 agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
     1081 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat
     1141 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg
     1201 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     1261 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact gcgatcgccc
     1321 gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     1381 cgtttagtga accgtcagat cactagaagc tttattgcgg tagtttatca cagttaaatt
     1441 gctaacgcag tcagtgcttc tgacacaaca gtctcgaact taagctgcag tgactctctt
     1501 aaggtagcct tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag
     1561 acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt
     1621 ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca
     1681 ctcccagttc aattacagct cttaaggcta gagtacttaa tacgactcac tataggctag
     1741 aatggtgtct aagggcgaag agctgattaa ggagaacatg cacatgaagc tgtacatgga
     1801 gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga
     1861 gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga
     1921 catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat
     1981 ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata
     2041 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     2101 ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa
     2161 aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg gcctggaagg
     2221 cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa acgccaagac
     2281 cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga
     2341 ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt
     2401 ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta attgaacgcg
     2461 tcgagcatgc atctagggcg gccaattccg cccctctccc ccccccccct ctccctcccc
     2521 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     2581 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     2641 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     2701 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     2761 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     2821 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     2881 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     2941 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     3001 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     3061 ttgaaaaaca cgatgataag cttgccacaa cccgggatcc tctagagtcg acccgggcgg
     3121 ccgcttccct ttagtgaggg ttaatgcttc gagcagacat gataagatac attgatgagt
     3181 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3241 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3301 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     3361 tctacaaatg tggtaaaatc cgataaggat cgatccgggc cccagctttt gttcccttta
     3421 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3481 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3541 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3601 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3661 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3721 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3781 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3841 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3901 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3961 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4021 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4081 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4141 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4201 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4261 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4321 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4381 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4441 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4501 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4561 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4621 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4681 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4741 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4801 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4861 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4921 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4981 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5041 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5101 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5161 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5221 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5281 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5341 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5401 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5461 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5521 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5581 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     5641 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5701 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5761 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5821 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5881 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5941 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     6001 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     6061 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     6121 ctattacgcc agtcgaccat agccaattca atatggcgta tatggactca tgccaattca
     6181 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
     6241 atatggtgga tctggacccc agccaattca atatggcgga cttggcacca tgccaattca
     6301 atatggcgga cttggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt
     6361 tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa
     6421 tggcggccat attggctata tgccaggatc aatatatagg caatatccaa tatggcccta
     6481 tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt
     6541 ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc
     6601 atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta
     6661 tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc
     6721 ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg
     6781 accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc
     6841 ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata
     6901 gtaacttggc aagtacatta ctattggaag gacgccaggg tacattggca gtactcccat
     6961 tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa
     7021 tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc
     7081 taatgggagg tctatataag caatgctcgt ttagggaac
//
