LOCUS Exported 6195 bp ds-DNA circular SYN 11-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBBR1MCS2-Tac-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6195)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sunday, September 11, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6195
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 458..1252
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
protein_bind 1657..1678
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1693..1723
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 1731..1747
/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)."
primer_bind 1755..1771
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 1792..1810
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind 1840..1856
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(1910..1987)
/gene="lacI"
/note="lacI promoter"
misc_feature 2033..2061
/note="tac promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
CDS 2108..2818
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/note="mCherry"
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
terminator 2852..2899
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(2941..2957)
/note="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(2990..3008)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3018..3034)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(3741..4403)
/codon_start=1
/product="replication protein for the broad-host-range
plasmid pBBR1 from Bordetella bronchiseptica"
/note="pBBR1 Rep"
/translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
rep_origin 4404..5173
/note="pBBR1 oriV"
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
ORIGIN
1 accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc
61 aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg
121 aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg
181 aaaagtgcca cctgggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
241 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt
301 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
361 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
421 aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
481 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
541 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
601 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
661 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
721 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
781 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
841 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
901 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
961 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
1021 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
1081 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
1141 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
1201 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
1261 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
1321 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
1381 atcctccagc gcggggatct catgctggag ttcttcgccc acccccatgg gcaaatatta
1441 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
1501 tggcttccat gtcggcagaa tgcttaatga attacaacag tttttatgca tgcgcccaat
1561 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
1621 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
1681 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
1741 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
1801 tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gaggtcgacg gtatcgataa
1861 gcttgatatc gaattcgagc tcggtacctc gcgaatgcat ctagatcaca ttcaccaccc
1921 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
1981 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa atttgacaat
2041 taatcatcgg ctcgtataat gtgtggaatg tctcatgagc ggatatttca cacaggaaac
2101 agtattcatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
2161 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
2221 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
2281 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
2341 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
2401 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
2461 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
2521 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
2581 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
2641 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
2701 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
2761 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaatt
2821 aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
2881 ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacacatcg
2941 gatccactag ttctagagcg gccgccaccg cggtggagct ccaattcgcc ctatagtgag
3001 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
3061 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
3121 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggaaattg
3181 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
3241 accaataggc cgactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
3301 ggtgccctta aacgcctggt gctacgcctg aataagtgat aataagcgga tgaatggcag
3361 aaattcgaaa gcaaattcga cccggtcgtc ggttcagggc agggtcgtta aatagccgct
3421 tatgtctatt gctggtttac cggtttattg actaccggaa gcagtgtgac cgtgtgcttc
3481 tcaaatgcct gaggccagtt tgctcaggct ctccccgtgg aggtaataat tgacgatatg
3541 atcatttatt ctgcctccca gagcctgata aaaacggtga atccgttagc gaggtgccgc
3601 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
3661 agacaaggta tagggcggcg aggcggctac agccgatagt ctggaacagc gcacttacgg
3721 gttgctgcgc aacccaagtg ctaccggcgc ggcagcgtga cccgtgtcgg cggctccaac
3781 ggctcgccat cgtccagaaa acacggctca tcgggcatcg gcaggcgctg ctgcccgcgc
3841 cgttcccatt cctccgtttc ggtcaaggct ggcaggtctg gttccatgcc cggaatgccg
3901 ggctggctgg gcggctcctc gccggggccg gtcggtagtt gctgctcgcc cggatacagg
3961 gtcgggatgc ggcgcaggtc gccatgcccc aacagcgatt cgtcctggtc gtcgtgatca
4021 accaccacgg cggcactgaa caccgacagg cgcaactggt cgcggggctg gccccacgcc
4081 acgcggtcat tgaccacgta ggccgacacg gtgccggggc cgttgagctt cacgacggag
4141 atccagcgct cggccaccaa gtccttgact gcgtattgga ccgtccgcaa agaacgtccg
4201 atgagcttgg aaagtgtctt ctggctgacc accacggcgt tctggtggcc catctgcgcc
4261 acgaggtgat gcagcagcat tgccgccgtg ggtttcctcg caataagccc ggcccacgcc
4321 tcatgcgctt tgcgttccgt ttgcacccag tgaccgggct tgttcttggc ttgaatgccg
4381 atttctctgg actgcgtggc catgcttatc tccatgcggt agggtgccgc acggttgcgg
4441 caccatgcgc aatcagctgc aacttttcgg cagcgcgaca acaattatgc gttgcgtaaa
4501 agtggcagtc aattacagat tttctttaac ctacgcaatg agctattgcg gggggtgccg
4561 caatgagctg ttgcgtaccc ccctttttta agttgttgat ttttaagtct ttcgcatttc
4621 gccctatatc tagttctttg gtgcccaaag aagggcaccc ctgcggggtt cccccacgcc
4681 ttcggcgcgg ctccccctcc ggcaaaaagt ggcccctccg gggcttgttg atcgactgcg
4741 cggccttcgg ccttgcccaa ggtggcgctg cccccttgga acccccgcac tcgccgccgt
4801 gaggctcggg gggcaggcgg gcgggcttcg ccttcgactg cccccactcg cataggcttg
4861 ggtcgttcca ggcgcgtcaa ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg
4921 ccttccactt ggtgtccaac cggcaagcga agcgcgcagg ccgcaggccg gaggcttttc
4981 cccagagaaa attaaaaaaa ttgatggggc aaggccgcag gccgcgcagt tggagccggt
5041 gggtatgtgg tcgaaggctg ggtagccggt gggcaatccc tgtggtcaag ctcgtgggca
5101 ggcgcagcct gtccatcagc ttgtccagca gggttgtcca cgggccgagc gaagcgagcc
5161 agccggtggc cgctcgcggc catcgtccac atatccacgg gctggcaagg gagcgcagcg
5221 accgcgcagg gcgaagcccg gagagcaagc ccgtagggcg ccgcagccgc cgtaggcggt
5281 cacgactttg cgaagcaaag tctagtgagt atactcaagc attgagtggc ccgccggagg
5341 caccgccttg cgctgccccc gtcgagccgg ttggacacca aaagggaggg gcaggcatgg
5401 cggcatacgc gatcatgcga tgcaagaagc tggcgaaaat gggcaacgtg gcggccagtc
5461 tcaagcacgc ctaccgcgag cgcgagacgc ccaacgctga cgccagcagg acgccagaga
5521 acgagcactg ggcggccagc agcaccgatg aagcgatggg ccgactgcgc gagttgctgc
5581 cagagaagcg gcgcaaggac gctgtgttgg cggtcgagta cgtcatgacg gccagcccgg
5641 aatggtggaa gtcggccagc caagaacagc aggcggcgtt cttcgagaag gcgcacaagt
5701 ggctggcgga caagtacggg gcggatcgca tcgtgacggc cagcatccac cgtgacgaaa
5761 ccagcccgca catgaccgcg ttcgtggtgc cgctgacgca ggacggcagg ctgtcggcca
5821 aggagttcat cggcaacaaa gcgcagatga cccgcgacca gaccacgttt gcggccgctg
5881 tggccgatct agggctgcaa cggggcatcg agggcagcaa ggcacgtcac acgcgcattc
5941 aggcgttcta cgaggccctg gagcggccac cagtgggcca cgtcaccatc agcccgcaag
6001 cggtcgagcc acgcgcctat gcaccgcagg gattggccga aaagctggga atctcaaagc
6061 gcgttgagac gccggaagcc gtggccgacc ggctgacaaa agcggttcgg caggggtatg
6121 agcctgccct acaggccgcc gcaggagcgc gtgagatgcg caagaaggcc gatcaagccc
6181 aagagacggc ccgag
//