LOCUS Exported 3628 bp ds-DNA circular SYN 03-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..3628
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 423..572
/label=HSP70 promoter
RBS 579..601
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 620..637
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 647..1366
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
rep_origin complement(1809..2397)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2568..3428)
/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(3429..3533)
/gene="bla"
/label=AmpR promoter
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 gcttgcatgc ctgcaggtcg
421 acgacccgca cgaccagcgt tagcatgctc agtaagttga gtgcatcagg ctcagctctg
481 aattgacagc acaccgccgt cgaggcaagc ttgagcgggg tgcactcatc atagtgcagg
541 aaagaagctc tacatattca ggaggattca ccggatcctt tgtttaactt taagaaggag
601 atataccatg ggcagcagcc atcatcatca tcatcacagc agcggcatgg tgagcaaggg
661 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
721 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
781 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
841 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
901 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
961 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
1021 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
1081 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
1141 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
1201 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
1261 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
1321 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaatttt tttataccgg
1381 gtaccgagct cgaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
1441 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
1501 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
1561 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
1621 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
1681 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
1741 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
1801 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
1861 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
1921 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
1981 tcgggaagcg tggcgctttc tcaaagctca cgctgtaggt atctcagttc ggtgtaggtc
2041 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
2101 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
2161 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
2221 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
2281 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
2341 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
2401 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
2461 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
2521 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
2581 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
2641 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
2701 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
2761 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
2821 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
2881 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
2941 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
3001 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
3061 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
3121 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
3181 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
3241 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
3301 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
3361 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
3421 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
3481 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
3541 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
3601 aataggcgta tcacgaggcc ctttcgtc
//