LOCUS Exported 3600 bp ds-DNA circular SYN 04-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..3600
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 9..27
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 59..81
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 101..118
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 122..154
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/label=T7 tag
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS 158..181
/codon_start=1
/product="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/label=Xpress tag
/translation="DLYDDDDK"
CDS 209..928
/codon_start=1
/product="Emerald GFP"
/label=EmGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
terminator 1004..1051
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 1148..1603
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1629..1733
/gene="bla"
/label=AmpR promoter
CDS 1734..2594
/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"
rep_origin 2765..3353
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta gaaataattt
61 tgtttaactt taagaaggag atatacatat gcggggttct catcatcatc atcatcatgg
121 tatggctagc atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa
181 ggatcgatgg ggatccgaat tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
241 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
301 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
361 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttgacctacg gcgtgcagtg
421 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
481 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
541 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
601 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaaggt
661 ctatatcacc gccgacaagc agaagaacgg catcaaggtg aacttcaaga cccgccacaa
721 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
781 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
841 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
901 tctcggcatg gacgagctgt acaagtaact cgagaagctt gatccggctg ctaacaaagc
961 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
1021 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatctgg
1081 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
1141 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
1201 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
1261 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
1321 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
1381 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
1441 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
1501 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
1561 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
1621 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
1681 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
1741 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
1801 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
1861 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
1921 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
1981 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
2041 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
2101 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
2161 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
2221 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
2281 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
2341 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
2401 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
2461 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
2521 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
2581 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
2641 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
2701 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
2761 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
2821 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
2881 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
2941 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
3001 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
3061 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
3121 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
3181 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
3241 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
3301 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
3361 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
3421 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
3481 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
3541 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagg atctcgatcc
//