LOCUS Exported 4281 bp ds-DNA circular SYN 24-MAY-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..4281
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(39..55)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 63..79
/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)."
promoter complement(87..117)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 132..153
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(441..1029)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1200..2060)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(2061..2165)
/gene="bla"
/label=AmpR promoter
primer_bind 2639..2655
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2722..3231
/label=35SPPDK promoter
/note="hybrid promoter consisting of the cauliflower mosaic
virus 35S enhancer fused to the maize C4PPDK basal promoter
(Yoo et al., 2007)"
misc_feature 3238..3290
/label=MCS
/note="multiple cloning site"
misc_feature 3264..3294
/label=MCS
CDS 3291..4010
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
terminator 4029..4281
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
ORIGIN
1 cgatgataag ctgtcaaaca tgagaattcg taatcatggt catagctgtt tcctgtgtga
61 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
121 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
181 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
241 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
301 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
361 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
841 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
1021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
1081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
1141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
1201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
1261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
1321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
1381 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
1441 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
1501 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
1561 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
1621 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
1681 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
1741 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
1801 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
1861 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
1921 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
1981 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
2041 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
2101 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
2161 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
2221 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
2281 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
2341 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
2401 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
2461 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
2521 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
2581 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
2641 aaaacgacgg ccagtgccaa gctctcgaga agcttactcc aagaatatca aagatacagt
2701 ctcagaagac caaagggcta ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct
2761 cggattccat tgcccagcta tctgtcactt catcaaaagg acagtagaaa aggaaggtgg
2821 cacctacaaa tgccatcatt gcgataaagg aaaggctatc gttcaagatg cctctgccga
2881 cagtggtccc aaagatggac ccccacccac aaggagcatc gtggaaaaag aagacgttcc
2941 aaccacgtct tcaaagcaag tggattgatg tgatatctcc actgacgtaa gggatgacgc
3001 acaatcccac tatccttcgc cccaagcttg ggcccaagct tgggtcgcgc cccacggatg
3061 gtataagaat aaaggcattc cgcgtgcagg attcacccgt tcgcctctca ccttttcgct
3121 gtactctctc gccacacaca ccccctctcc agctccgttg gagctccgga cagcagcagg
3181 cgcggggcgg tcacgtagta agcagctctc ggctccctct ccccttgctc cgtggatctc
3241 gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc atggtgagca
3301 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
3361 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
3421 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
3481 ccttcaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
3541 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
3601 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
3661 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
3721 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
3781 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
3841 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
3901 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
3961 tcgtgaccgc cgccgggatc actcacggca tggacgagct gtacaagtaa agcggccgcc
4021 cggctgcaga tcgttcaaac atttggcaat aaagtttctt aagattgaat cctgttgccg
4081 gtcttgcgat gattatcata tatatttctg ttgattacgt taagcatgta ataattaaca
4141 tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg caattataca
4201 tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg
4261 tgtcatctat gttactagat c
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