LOCUS Exported 6470 bp ds-DNA circular SYN 12-JUL-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pMIR-Reporter
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6470)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sunday, September 4, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6470
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 379..395
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(551..2200)
/codon_start=1
/gene="luc"
/product="firefly luciferase"
/note="luciferase"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIVPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
promoter complement(2225..2428)
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
enhancer 2429..2732
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter complement(2854..2872)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(2884..2902)
/note="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2922..2938)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2946..2962
/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(2970..3000)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3015..3036
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3324..3912)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4083..4943)
/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(4944..5048)
/gene="bla"
/note="AmpR promoter"
polyA_signal 5136..5217
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(5447..6046)
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="PuroR"
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
promoter complement(6143..6459)
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 6157..6292
/note="SV40 ori"
/note="SV40 origin of replication"
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctagcggcc gcatacaaaa
421 aaccaacaca cagatccaat gaaaataaaa gatcctttat taagcttaac tgtttaaacc
481 tatgatatca cgcacgcgtc tatgccggcc tatgagctcc ctcactagtg cgcagctttc
541 atcactcgag caatttggac tttccgccct tcttggcctt tatgaggatc tctctgattt
601 ttcttgcgtc gagttttccg gtaagacctt tcggtacttc gtccacaaac acaactcctc
661 cgcgcaactt tttcgcggtt gttacttgac tggcgacgta atccacgatc tctttttccg
721 tcatcgtctt tccgtgctcc aaaacaacaa cggcggcggg aagttcaccg gcgtcatcgt
781 cgggaagacc tgccacgccc gcgtcgaaga tgttggggtg ttgtaacaat atcgattcca
841 attcagcggg ggccacctga tatcctttgt atttaattaa agacttcaag cggtcaacta
901 tgaagaagtg ttcgtcttcg tcccagtaag ctatgtctcc agaatgtagc catccatcct
961 tgtcaatcaa ggcgttggtc gcttccggat tgtttacata accggacata atcataggtc
1021 ctctgacaca taattcgcct ctctgattaa cgcccagcgt tttcccggta tccagatcca
1081 caaccttcgc ttcaaaaaat ggaacaactt taccgaccgc gcccggttta tcatccccct
1141 cgggtgtaat cagaatagct gatgtagtct cagtgagccc atatccttgt cgtatccctg
1201 gaagatggaa gcgttttgca accgcttccc cgacttcttt cgaaagaggt gcgcccccag
1261 aagcaatttc gtgtaaatta gataaatcgt atttgtcaat cagagtgctt ttggcgaaga
1321 atgaaaatag ggttggtact agcaacgcac tttgaatttt gtaatcctga agggatcgta
1381 aaaacagctc ttcttcaaat ctatacatta agacgactcg aaatccacat atcaaatatc
1441 cgagtgtagt aaacattcca aaaccgtgat ggaatgggac aacacttaaa atcgcagtat
1501 ccggaacgat ttgattgcca aaaataggat ctctggcatg cgagaatctg acgcaggcag
1561 ttctatgcgg aagggccaca cccttaggta acccagtaga tccagaggaa ttcattatca
1621 gtgcaattgt tttgtcacga tcaaaggact ctggtacaaa atcgtattca ttaaaaccgg
1681 gaggtagatg agatgtgacg aacgtgtaca tcgactgaaa tccctggtaa tccgttttag
1741 aatccatgat aataattttc tggattattg gtaatttttt ttgcacgttc aaaatttttt
1801 gcaacccctt tttggaaaca aacactacgg taggctgcga aatgttcata ctgttgagca
1861 attcacgttc attataaatg tcgttcgcgg gcgcaactgc aactccgata aataacgcgc
1921 ccaacaccgg cataaagaat tgaagagagt tttcactgca tacgacgatt ctgtgatttg
1981 tattcagccc atatcgtttc atagcttctg ccaaccgaac ggacatttcg aagtattccg
2041 cgtacgtgat gttcacctcg atatgtgcat ctgtaaaagc aattgttcca ggaaccaggg
2101 cgtatctctt catagcctta tgcagttgct ctccagcggt tccatcctct agaggataga
2161 atggcgccgg gcctttcttt atgtttttgg cgtcttccat ggtggatcca cggttcacta
2221 aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
2281 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
2341 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
2401 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
2461 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
2521 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
2581 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
2641 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
2701 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
2761 atgaccccgt aattgattac tattaataac taagatctgg taccttgaat tcatgcttct
2821 cctcccttta gtgagggtaa ttctctctct ctccctatag tgagtcgtat taattccttc
2881 tcttctatag tgtcacctaa atcgttgcaa ttcgtaatca tgtcatagct gtttcctgtg
2941 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
3001 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
3061 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
3121 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
3181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
3241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
3301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
3361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
3421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
3481 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
3541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
3601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
3661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
3721 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
3781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
3841 aaaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
3901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
3961 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
4021 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
4081 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
4141 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
4201 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
4261 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
4321 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
4381 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
4441 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
4501 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
4561 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
4621 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
4681 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
4741 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
4801 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
4861 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
4921 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
4981 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
5041 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
5101 acattaacct ataaaaatag gcgtatcacg agattgcagt gaaaaaaatg ctttatttgt
5161 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
5221 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
5281 agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga gtcggtgggc
5341 ctcgggggcg ggtgcggggt cggcggggcc gccccgggtg gcttcggtcg gagccatggg
5401 gtcgtgcgct cctttcggtc gggcgctgcg ggtcgtgggg cgggcgtcag gcaccgggct
5461 tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga
5521 agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg aaggcgggca
5581 ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc agacccttgc
5641 cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt
5701 gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg ctcaactcgg
5761 ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc
5821 agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg agcccgacgc
5881 gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc gggtcgacgg
5941 tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg gcccggggga
6001 cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggaa ggtcgtctcc
6061 ttgtgagggg tcaggggcgt gggtcagggg atggtggcgg caccggtcgt ggcggccgac
6121 ctgcaggcat gcaagctagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
6181 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta
6241 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
6301 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
6361 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca
6421 tacttctgcc tgctggggag cctggggact ttccacacca atctttcgtc
//